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Male Factor Infertility
Male Factor Infertility can result from abnormalities in sperm
production including low sperm count, low motility, poor
morphology, anti-sperm antibodies, and hyper-coagulation /
viscosity of the seminal fluid. Chinese medicine has been very
effective at helping to resolve all of these male issues.
Many studies have been done in China and in the U.S. that show
Chinese Medicine and acupuncture improve the sperm count,
morphology, motility and help the anti-sperm antibodies too. See
the Medical Research for male Factor infertility.
Chinese medicine treatments include acupuncture, herbs,
anti-oxidants and acupressure, which improve the male’s
reproductive capacity.
Male Factor Infertility Articles
General Articles
Male Factor Infertility
In the United States, it is estimated that approximately 15% of
the population falls into the category of being unable to
conceive. In 40% of these cases, sperm abnormalities are either
a factor or the factor...
Medical Research Articles
Study: Acupuncture May Improve Fertility in Men
In many cultures, women are unfairly blamed for the inability of
a sexually active couple to conceive. In reality, men suffer
from infertility issues just as frequently as women. According
to statistics from the National Infertility Association (an
organization also known as RESOLVE),
Acupuncture May
Improve Sperm Quality
Acupuncture may help some men overcome infertility problems by
improving the quality of their sperm, according to a new
study...
Effect of acupuncture on sperm parameters of males suffering
from subfertility related to low sperm quality.
The aim of this prospective controlled study was to assess the
effect of acupuncture on the sperm quality of males suffering
from subfertility related to sperm impairment. Semen samples of
16 acupuncture-treated subfertile patients were analyzed before
and 1 month after treatment (twice a...
Effects of guizhi-fuling-wan on male infertility with
varicocele.
Thirty-seven infertile patients with varicocele were treated
with Guizhi-Fuling-Wan (7.5 g/day) for at least 3 months. Before
and after the administration, semen qualities such as sperm
concentration and motility were examined, and the varicocele was
graded. A varicocele disappearance rate of...
Eighty-seven cases of male infertility treated by bushen
shengjing pill in clinical observation and evaluation on its
curative effect
Eighty-seven cases of male infertility with semen abnormality
were treated and observed by Bushen Shengjing Pill, its curative
effect was evaluated with quantitative assessment and analytical
comparison comprehensive scoring of semen routine analysis.
These patients were treated for one...
An experimental study on inhibitory effect of Chinese medicine
tai-bao on antisperm antibody
OBJECTIVE: To investigate whether Chinese medicine Tai-bao could
inhibit antisperm antibody in experimental mice. METHODS: The
experimental immunoinfertility mice were due to antisperm
antibody induced by injection of human sperm membrane antigens.
The experimental immuno-
Male Factor Infertility
Randine Lewis, Ph.D., Lic.Ac.
In the United States, it is estimated that approximately 15% of
the population falls into the category of being unable to
conceive. In 40% of these cases, sperm abnormalities are either
a factor or the factor.
Male factor infertility is assessed based upon the following
values:
deficient sperm count (less than 10 million per millileter;
volume should be 1 - 5 mL of ejaculate)
insufficient sperm motility (over 60% should be motile and
demonstrate purposeful forward movement), and/or
poor sperm morphology (more than 50-60% abnormal in form)
Infertility is defined as the inability to fertilize the ovum;
whereas sterility is defined as the lack of sperm production.
The average ejaculate sample contains almost 200 million sperm.
Amazingly enough, only a few dozen sperm actually reach the egg
for a chance at penetration. This makes for some pretty ominous
statistics for sperm overall. It is for this reason that sperm
numbers must be so high, just to have a modicum of hope of
reaching the vicinity of the egg traveling down the fallopian
tube. If both partners have fertility issues, it seems truly a
miracle that conception ever even takes place. Luckily, there
are methods to improve sperm count, motility, and morphology.
Etiology
Male fertility depends upon adequate production of spermatozoa
by the testes, unobstructed transit of sperm through the seminal
tract, and satisfactory delivery to the ovum. Deficient sperm
production may be affected by factors such as radiation and
other environmental toxins, undescended testis, varicocele,
traumatic induced or infectious testicular atrophy, drug
effects, prolonged fever, and endocrine disorders that affect
the hypothalamic-pituitary-gonadal axis. Antisperm antibodies
may be a factor in certain couples, and may be produced by
either partner. If a man produces antibodies to his own sperm,
the antibodies will typically attack the sperm's tail. If the
woman produces sperm antibodies, they will often attack the head
of the sperm.
Congenital anomalies may obstruct the seminal tract, as well as
certain surgical procedures. Low sperm counts can be aggravated,
if not caused, by factors such as tight fitting underwear which
raises the scrotal temperature, environmental toxins, urogenital
infections, poor diet and prescription drugs (anti-hypertensives
and anti-inflammatories can drastically reduce sperm count).
Even anti-histamines negatively affect sperm count, by
diminishing the seminal fluid, which contains high levels of
anti-oxidants within it. Stress, lack of sleep, and overuse of
alcohol, nicotine and marijuana decrease sperm production as
well.
When the cause of the abnormality is known, often its
identification and elimination can cure the problem. In other
cases, deeper analysis is necessary.
Diagnosis
Significant medical history would include a history of childhood
cryptorchidism (failure of the testes to descend), mumps, or
history of sexual problems. Physical manifestations may include
structural abnormalities, particularly the presence of a
varicocele (scrotal swelling). The size and shape of the
testicles should be within the normal range. General evaluation
of secondary sex characteristics may provide clues to an
underlying endocrine disorder. Hypothyroidism, hypopituitarism,
other functional adrenal disorders, and hypogonadism are certain
endocrine disorders which may possibly play a role in sperm
abnormalities.
Male sterility is easier to diagnose with western methods than
female infertility, but harder to treat. The only potential
remedy is surgery. Yet many men with sperm problems are treated
effectively with nutritional suplementation and herbs. If the
physical examination reveals no abnormality and the man is not
impotent (able to engage in intercourse, can become erect, and
can ejaculate), the next diagnostic step consists of obtaining a
sperm specimen and examining the ejaculate histologically for
numbers, motility, and morphology (correct shape). A minimum of
2 to 3 specimens should be analyzed before determining ejaculate
adequacy, as sperm values can fluctuate from one sample to the
next.
Grossly the semen should look slightly viscous and opaque, and
the volume should be between 1 and 5 mL.
Sperm density should be (optimally) over 20 million/mL. The
results of semen analyses are recorded into the following
categories:
adequate
aspermia - absence of ejaculate (surgical sequelae or neurogenic
dysfunction)
azoospermia - absence of sperm in the semen (from testicular
disorders)
oligospermia - lowered sperm density
diminished motility and impaired sperm forward progression
abnormal sperm morphology
antisperm antibodies.
An Overview of Sperm Production
Sperm production begins during puberty in response to the same
hormones (LH and FSH) as in the female. But the LH signals cells
within the leydig cells of the testes to produce testosterone,
and FSH signals sertoli cells to produce sperm. Estrogen is also
important in sperm formation, but too much dietary synthetic
sources of estrogen can be harmful.
The seminal vesicles secrete substances which nourish the sperm,
including fructose (which feeds the sperm), fibrinogen (which
holds or coagulates the fluid together) and prostaglandins
(which help the sperm penetrate the cervix). The prostate adds
an alkaline fluid to the ejaculate. It is extremely important to
keep the sperm in a more alkaline environment because the
vaginal pH is relatively acidic. Seminal fluid in normal,
fertile men contains antioxidant factors. In many subfertile men
the seminal fluid may not contain the protective elements, or
the circulating free radicals may be so abundant that the
seminal fluid is not capable of scavenging the damaged reactive
oxygen species. Therefore, men with suboptimum sperm counts
should include dietary sources of antioxidants.
The plasma membrane of human sperm contains high levels of
polyunsaturated fatty acids, making them extremely susceptible
to peroxidative changes. Free radical damage leads to functional
impairment in the sperm, lowering motility and morphology.
Most vaginal lubricants are hostile to sperm. The only vaginal
lubricants which have been found to support sperm longevity are
egg whites (yes, really) and canola oil.
Treatment
Avoid excess environmental toxins including synthetic estrogens.
Beef and dairy cattle are often fed bovine growth hormone to
enhance growth and milk production. Most meat, dairy products,
and even poultry and eggs contain substantial quantities of
synthetic estrogens. Some reports have shown the presence of
synthetic estrogen in sources of drinking water as well.
Therefore, purified drinking water is suggested.
Pesticides and other chemicals which may impair spermatogenesis
are found in non-organically grown produce. It is therefore best
to consume organic fruits and vegetables.
Keep scrotal temperatures between 94 and 96 degrees Farenheit.
Men with slight varicoceles are encouraged to use cool packs
daily on the testicles.
Avoid saturated fats, hydrogenated oils, coconut, palm and
especially cottonseed oil (contains gossypol which inhibits
sperm formation).
Include polyunsaturated oils and essential fatty acids.
Natural Supplements
Soy products contain isoflavones or phytoestrogens which occupy
estrogen receptor sites at the exclusion of circulating
synthetic estrogens, and have a very weak estrogenic (which
physiologically translates to anti-estrogenic) effect. Soy,
other legumes, nuts and seeds also contain phytosterols which
promote testosterone production.
Oxidative damage is present in almost half of the diagnosed
cases of oligospermia. To prevent further free radical damage to
developing sperm, it is recommended that the following
nutritional supplementation be included:
Vitamin C - 2,000 mg/day (in divided doses)
Vitamin E - 800 IU/day
Beta-carotene - 100,000 IU/day
Selenium
Other nutritional supplements which are critical to sperm
production include:
Zinc - 60 mg/day (necessary for sperm production and
testosterone metabolism)
Vitamin B12 - 1000 ug/day (involved in the replication of cells)
L-Arginine - 4 g/day (an amino acid involved in cellular
replication)
L-Carnitine - 600 mg. three times per day (found in very high
levels in sperm, this amino acid transports fatty acids into the
mitochondria and assists sperm motility)
Because of sperm's susceptibility to oxidative damage it is
recommended to include free-radical scavengers like oligomeric
proanthocyanidins. One of the most potent bioactive antioxidant
sources comes from the extracts of pine bark extract, red wine
extract, grape seed extract, and bilberry extract. Oligomeric
proanthocyanidins may be purchased through health and
nutritional sources.
TCM Diagnosis
From a Chinese perspective, the main causes of male infertility
fall under two broad categories: one is a deficiency of the
Kidneys (usually kidney Yang; sometimes kidney yin); the other
is damp-heat in the pelvic organs. [Kidney deficiency may also
affect the liver and spleen and lead to stasis of qi and blood.]
The presence of a varicocele translates to blood stasis in our
Chinese medical diagnosis. The swollen veins obstruct transit;
it is therefore necessary to invigorate and move the blood so
the sperm can develop normally.
Chinese Medical Treatment
Ginseng (Chinese, Korean, or Siberian), which supplements the
source qi, promotes testicular growth, testosterone levels and
sperm formation.
Cornus Officinalis Fructus, used to stabilize the kidney
essence, and tonify the liver and kidneys, has been found to
improve sperm motility.
Kidney yang tonics like Eucommia, Epimedii, Radix Morindae
Officinalis and Cornu Cervi Parvum are used in the appropriate
presentation of impotence, fatigue, low back pain, urinary
frequency and spermatorrhea.
Sperm antibodies are addressed according to pattern
discrimination, for both males and females, and treated
accordingly.
Most men with diagnosed varicocele that I treat respond to
improvement with the formula Cinnamon and Poria decoction or Gui
Zhi Fu Ling Wan, which consists of Ramulus Cinnamomi Cassiae,
Sclerotium Poriae Cocos, Radis Paeoniae, Cortex Moutan Radicis,
and Semen Persicae. This formula, which is traditionally used
for gynecologic disorders of blood stasis in the uterus, has
proven very promising in treating morphologic sperm
abnormalities resulting from varicocele. The formula invigorates
the blood, inhibiting the pooling mechanism which causes the
poor sperm quality. A study from the American Journal of Chinese
Medicine, 24, 1996, on The Effects of Guizhi-fuling-wan on male
infertility with varicocele was conducted by Ishikawa, Ohashi,
Hayakawa, Kaneko & Hata at the Department of Urology, Ichikawa
General Hospital in Japan. The abstract reported that 37
infertile patients with varicocele were treated with Gui Zhi Fu
Ling Way, (7.5 g/day) for three months. Semen qualities such as
sperm concentration and motility were graded. A varicocele
disappearance rate of 80% was obtained with 40 out of 50
varicoceles, and sperm count and motility improvements were
found in 71.4% and 62.1% of patients, respectively.
Journal of Chinese Medicine, Number 54, May 1997, entitled Xu
Runsan's Experience in Treating Sperm Abnormality, stated the
main causes of sperm abnormality are deficiency of the kidney
yang or kidney yin, or deficiency of the kidneys which affects
the liver and spleen and leads to stasis of qi and blood or
downward flow of damp-heat.
Differentiation and treatment was made as follows:
1) Deficiency of kidney yang
aversion to cold
low back pain
coldness in the scrotum
deep and thready pulse
thin and white tongue coating
You Gui Wan
Shu Di Huang, Shan Yao, Shan Zhu Yu, Tu Si Zi, Gou Qi Zi, Lu
Jiao Jiao, Du Zhong, Dang Gui, Rou Gui, Fu Zi
for patients with aspermia remove Du Zhong, Rou Gui and Fu Zi
and add Chuan Xiong and Hong Shen
for patients with absence of sperm liquefaction add Bei Xie
for patients with dead sperm add Xu Duan
Giovanni Maciocia's Obstetrics & Gynecology in Chinese Medicine
suggests treating kidney yang deficiency with the prescription:
Wu Zi Yan Zong Wan, Five Seeds Developing the Ancestors Pill:
Lycium, Cuscutta, Schissandra, Semen Plantaganis, and Fructus
Rubrus.
2) Deficiency of kidney yin
emaciation
irritability
weak, frail pulse
red tongue body
Zuo Gui Wan variation
Shu Di Huang, Shan Yao, Shan Zhu Yu, Tu Si Zi, Gou Qi Zi, Gui
Jiao, Lu Jiao Jiao, Niu Xi
for patients with aspermia add Dang Gui, Chuan Xiong, Nu Zhen Zi,
and Han Lian Cao
for patients with absence of sperm liquefaction add Dan Shen,
Bei Xie, and Huang Bai
Stimulate acupuncture points
Sp 6 Three yin meeting
Ren 4
K3
K7
A study conducted by the College of Acupuncture & Moxabustion at
the Shanghai University of TCM, Shanghai, China, reported 35
cases of dysspermia infertility were treated only with low
frequency electroacupuncture on Sp6, Ren 12 and Ren 4 along with
moxibustion (heating the acupoints). The results of the study
showed improvement in lumbosacral aching, frequent urination,
emission and prospermia; activity and quantity of sperm, semen
quality and spermatogenic environment (semen quantity increased
obviously after treatmetn with significant decreasae of mucosity
and liquefaction time) improved. Sex hormones were normalized as
follows:
33.5% improvement in FSH
35.3% in LH
57.1% in estrogen
65.1% in testosterone
Study:
Acupuncture May Improve Fertility in Men
In many cultures, women are unfairly blamed for the inability of
a sexually active couple to conceive. In reality, men suffer
from infertility issues just as frequently as women. According
to statistics from the National Infertility Association (an
organization also known as RESOLVE), between 35 percent and 40
percent of infertility problems among couples are actually
caused by male conditions. Several factors may be responsible
for male infertility, including low sperm count, abnormal sperm
shape and size, and reduced motility. Lifestyle, genetics, and
physiological changes can also raise or lower male fertility
levels, and can significantly affect a man's ability to produce
offspring.
Previous research has shown that acupuncture can improve
fertility levels in women. Fewer studies on male infertility
have been conducted, although evidence suggests that acupuncture
can have an effect on sperm production and quality, without
causing any changes in behavior or sexual desire.
A recent trial published in Fertility and Sterility has shown
just how effective acupuncture can be in the treatment of this
condition, leading to significant increases in the number of
normal sperm and equally significant reductions in structural
defects.
In the study, 28 men who were diagnosed with idiopathic
infertility received acupuncture twice a week over a period of 5
weeks. The following acupuncture points were used as main
points: Guan yuan (Ren 4), shen shu (UB 23, bilateral), ci liao
(UB 32, bilateral), tai cong (Liv 3, bilateral), and tai xi (KI
3, bilateral). Secondary points included zhu san li (ST 36,
bilateral), xue hai (SP 10, bilateral), san yin jiao (SP 6,
bilateral), gui lai (ST 29, bilateral), and bai hui (Du 20).
Needles were inserted to a depth of between 15 and 25
millimeters, depending on the region of the body being treated.
Needles were manipulated for 10 minutes to achieve de qi, then
left in place for another 25 minutes before being removed.
Semen samples were collected from each of the men after a 3-day
period of sexual abstinence. Two samples were collected from
each patient: one obtained the day before treatment began, the
other after the last acupuncture treatment. Samples from the
treatment group were then randomized with semen samples from 12
untreated control patients and analyzed.
Compared to the control group, motility levels increased
significantly in semen samples in the men receiving acupuncture.
While median motility levels increased from 32% to 37% in the
control group, they increased from 44.5% to 50% in the
acupuncture group.
The number and percentage of healthy sperm also increased
dramatically in the acupuncture patients. At baseline, only
0.06% the sperm among men in the acupuncture group was
considered "healthy," while the median number of healthy sperm
calculated in ejaculate was 0.04 x 10 6 (40,000). After 10
sessions of treatments, the median percentage of healthy sperm
had increased more than four-fold, to 0.26%, while the median
number of healthy sperm per sample had reached 0.2 x 10 6
(200,000).
In addition, significant changes in sperm structure and quality
were seen in the samples from the acupuncture group. Before
treatment, only 22.5% of the sperm samples in the acupuncture
patients contained normal-shaped acrosomes, a cap-like structure
that develops over the anterior portion of a sperm cell's
nucleus. After treatment, the median percentage of normal
acrosome shapes showed a "statistically significant improvement"
to 38.5%.
Similarly, the percentage of sperm with a normal axoneme pattern
increased significantly among men receiving acupuncture. (The
axoneme is a microscopic structure that contains a series of
tubules arranged in a distinct pattern, and is believed to aid
in sperm motility.) Prior to the start of the study, the correct
axoneme pattern was present in 52% of sperm in the control
group, but only 46.1% in the acupuncture group. After 5 weeks of
therapy, the median percentage increased to 52.2% in acupuncture
patients, but actually decreased to 38.2% in the control group.
While acupuncture appeared able to improve the overall quality
and structural integrity of sperm, it was ineffective against
some common sperm pathologies. Apoptosis levels (programmed cell
death) in sperm samples were reduced slightly, but not to a
statistically significant degree. Median percentages of necrosis
(unprogrammed cell death) and sperm immaturity also decreased
slightly in the acupuncture group, but not to a level considered
statistically significant.
The authors concluded that despite the inability of acupuncture
to significantly reduce some sperm abnormalities, the treatment
could be used to improve overall sperm quality, leading to the
possibility of increased fertility.
"In conjunction with ART or even for reaching natural fertility
potential, acupuncture treatment is a simple, noninvasive method
that can improve sperm quality," the authors concluded. "Further
research is needed to demonstrate what stages and times in
spermatogenesis are affected by acupuncture, and how acupuncture
causes the physiologic changes in spermatogenesis."
References
Hopps CV, Goldstein M. Male infertility: the basics.
Available online at:
http://my.webmd.com/content/article/71/81282.htm.
Levine D. Boxers or briefs: myths and facts about men's
infertility.
Available online at:
http://my.webmd.com/content/article/11/1687_50040.htm.
Pei J, Strehler E, Noss U, et al. Quantitative evaluation of
spermatozoa ultrastructure after acupuncture treatment for
idiopathic male infertility. Fertility and Sterility July 2005;
84(1):141-7.
Acupuncture May Improve Sperm Quality
Study Shows Alternative Treatment May Help Male Infertility
Problems
By
Jennifer Warner
WebMD Medical News
Reviewed By
Brunilda Nazario, MD
on Wednesday, August 03, 2005
Aug. 3, 2005 -- Acupuncture may help some men overcome
infertility problems by improving the quality of their sperm,
according to a new study.
Researchers found five weeks of acupuncture treatment reduced
the number of structural abnormalities in sperm and increased
the overall number of normal sperm in a group of men with
infertility problems.
They say the results suggest that acupuncture may complement
traditional infertility treatments and help men reach their full
reproductive potential.
Acupuncture May Ease Male Infertility
An estimated 10% of men are infertile, and the male partner is a
factor in up to 50% of infertile couples, write the researchers.
In many cases, the cause of male infertility is unknown.
Previous studies of acupuncture and male infertility have
suggested that acupuncture can improve sperm production and
motility (a measure of sperm movement).
In this study, researchers looked at the effects of acupuncture
on the structural health of sperm in men with infertility of
unknown cause. The findings appear in the July issue of
Fertility and Sterility.
Twenty-eight infertile men received acupuncture treatments twice
a week for five weeks, and 12 received no treatment and served
as a comparison group.
Researchers analyzed sperm samples at the beginning and end of
the study and found significant improvements in sperm quality in
the acupuncture group compared with the other group.
Acupuncture treatment was associated with fewer structural
defects in the sperm and an increase in the number of normal
sperm in ejaculate.
But other sperm abnormalities, such as immature sperm or sperm
death, were unaffected by acupuncture.
The researchers write that acupuncture treatment is a simple,
noninvasive method that can improve sperm quality.
SOURCE: Pei, J. Fertility and Sterility, July 2005; vol 84: pp
141-147.
Effect of acupuncture on sperm parameters of males suffering
from subfertility related to low sperm quality.
Siterman S; Eltes F; Wolfson V; Zabludovsky N; Bartoov B
Institute of Chinese Medicine, Tel Aviv, Israel.
The aim of this prospective controlled study was to assess the
effect of acupuncture on the sperm quality of males suffering
from subfertility related to sperm impairment. Semen samples of
16 acupuncture-treated subfertile patients were analyzed before
and 1 month after treatment (twice a week for 5 weeks). In
parallel, semen samples of 16 control untreated subfertile males
were examined. Two specimens were taken from the control group
at an interval of 2-8 months. The expanded semen analysis
included routine and ultramorphological observations. The
fertility
index increased significantly (p < or = .05) following
improvement in total functional sperm fraction, percentage of
viability, total motile spermatozoa per ejaculate, and integrity
of the axonema (p < or = .05), which occurred upon treatment.
The intactness of axonema and sperm motility were highly
correlated (corr. = .50, p < or = .05). Thus, patients
exhibiting a low fertility potential due to reduced sperm
activity may benefit from acupuncture treatment.
Effects of guizhi-fuling-wan on male infertility with varicocele.
Ishikawa H; Ohashi M; Hayakawa K; Kaneko S; Hata M
Department of Urology, Ichikawa General Hospital, Tokyo Dental
College, Chiba, Japan.
Thirty-seven infertile patients with varicocele were treated
with Guizhi-Fuling-Wan (7.5 g/day) for at least 3 months. Before
and after the administration, semen qualities such as sperm
concentration and motility were examined, and the varicocele was
graded. A varicocele disappearance rate of 80% was obtained with
40 out of 50 varicocele, and improvement of sperm concentration
and motility were found in 71.4% and 62.1% of patients,
respectively. From these results, Guizhi-Fuling-Wan is
considered to be effective for circulation disorders in
varicocele as well as semen quality
Eighty-seven cases of male infertility treated by bushen
shengjing pill in clinical observation and evaluation on its
curative effect
Yue GP; Chen Q; Dai N
Institute of Acupuncture and Meridians, Anhui College of TCM,
Hefei.
Eighty-seven cases of male infertility with semen abnormality
were treated and observed by Bushen Shengjing Pill, its curative
effect was evaluated with quantitative assessment and analytical
comparison comprehensive scoring of semen routine analysis.
These patients were treated for one of three courses of
treatment, the semen quality was enhanced obviously, the
comprehensive semen routine analysis score was enhanced
significantly (P < 0.001) as compared with that before
treatment, the spouse pregnant rate was 56.32% (49/87), and
total effective rate was 95.40%
(83/87).
The result showed that this prescription had bidirectional
regulatory function in folliclestimulating hormone, luteotropic
hormone, testosterone, corticosterone, and could make the
enhanced or reduced hormone level to normal value.
An experimental study on inhibitory effect of Chinese medicine
tai-bao on antisperm antibody
Lai AN; Song JF; Liu XJ
Xiyuan Hospital, China Academy of TCM, Beijing.
OBJECTIVE: To investigate whether Chinese medicine Tai-bao could
inhibit antisperm antibody in experimental mice. METHODS: The
experimental immunoinfertility mice were due to antisperm
antibody induced by injection of human sperm membrane antigens.
The experimental immuno-infertile mice used in the present study
were divided into four groups including Tai-bao high dose group
(46.8 g.kg-1.d-1), Tai-bao low dose group (31.2 g.kg-1.d-1),
prednisone group and normal
saline group. The enzyme linked immune sorbent assay (ELISA) and
microcytotoxic assay were used for detection of antisperm
antibody. The change of levels of antisperm antibody before and
after treatment, pregnant rate, and the number of implantation
were investigated in tested mice. RESULTS: The pregnant rates in
normal saline group, prednisone group, Tai-bao high dose group
and low dose were 38.89%, 47.06%, 70.00% and 75.00%
respectively. The rate of pregnancy in Tai-bao low dose group
was significantly higher as compared with normal saline group (P
< 0.05). The rate of implantation in Tai-bao low dose group was
significantly higher than that in prednisone group (P < 0.05).
The results of detection of cytotoxic antibody to sperm showed
that cytotoxic percentages in Tai-bao high dose group (63.0 +/-
10.3%) and prednisone group (56.3 +/- 13.7%) were significantly
lower (P < 0.05 and P < 0.01) than that in normal saline group
(72.84 +/- 5.05%). CONCLUSION: Chinese medicine Tai-bao
possesses regulatory effect on reproductive immune function,
inhibitory effect on antisperm cytotoxic antibody, and promoting
effect on pregnancy.
Role of acupuncture in the treatment of female infertility
Raymond Chang, M.D.[a,b] Pak H. Chung, M.D.[b] and Zev Rosenwaks,
M.D.[c]
The Institute of East-West Medicine and the Center for
Reproductive Medicine and Infertility, Weill Medical College of
Cornell University, New York, New York
FERTILITY AND STERILITY® VOL. 78, NO. 6, DECEMBER 2002
Copyright ©2002 American Society for Reproductive Medicine
Published by Elsevier Science Inc., Printed on acid-free paper
in U.S.A.
Objective: To review existing scientific rationale and clinical
data in the utilization of acupuncture in the treatment of
female infertility.
Design: A MEDLINE computer search was performed to identify
relevant articles.
Result(s): Although the understanding of acupuncture is based on
ancient medical theory, studies have suggested that certain
effects of acupuncture are mediated through endogenous opioid
peptides in the central nervous system, particularly
ß-endorphin. Because these neuropeptides influence gonadotropin
secretion through their action on GnRH, it is logical to
hypothesize that acupuncture may impact on the menstrual cycle
through these neuropeptides. Although studies of adequate
design, sample size, and appropriate control on the use of
acupuncture on ovulation induction are lacking, there is only
one prospective randomized controlled study examining the
efficacy of acupuncture in patients undergoing IVF. Besides its
central effect, the sympathoinhibitory effects of acupuncture
may impact on uterine blood flow.
Conclusion(s): Although the definitive role of acupuncture in
the treatment of female infertility is yet to be established,
its potential impact centrally on the
hypothalamic-pituitary-ovarian axis and peripherally on the
uterus needs to be systemically examined. Prospective randomized
controlled studies are needed to evaluate the efficacy of
acupuncture in the female fertility treatment. (Fertil Steril®
2002;78:1149-53. ©2002 by American Society for Reproductive
Medicine.)
Key Words: Acupuncture, female infertility, in vitro
fertilization
Acupuncture as a therapeutic intervention has been extensively
studied and is increasingly practiced in the United States. A
recent survey of acupuncture released by an NIH Consensus
Development panel (1) indicated that although there are inherent
problems of design, sample size, and appropriate controls in the
acupuncture literature, promising data exist for the use of
acupuncture in treating nausea and vomiting (2), postoperative
pain (3-5), addiction (6-9), and general pain syndromes (10-12).
As a medical technique, acupuncture has also been reported as an
adjunct in the treatment of various gynecologic problems
(13-15).
Although conventional treatment options for female infertility
have been well established, there have been few systematic
reviews of complementary or alternative approaches to the
treatment of infertility. In light of an increasing trend in the
use of complementary and alternative medicine (16) and common
inquiry and utilization of such approaches by patients suffering
from infertility, we intend to review the existing scientific
rationale and clinical data based on which acupuncture may exert
an influence on the outcome of female fertility.
In examining the potential usefulness of acupuncture in
enhancing female fertility, it is appropriate first to give some
theoretical background for acupuncture. Although the theory of
acupuncture stems from underlying traditional Chinese medicine
premises that would define etiologies for infertility in terms
of energy disturbance of imbalances, or organ deficiencies and
excesses, we intend to review the existing literature by
examining modern medical aspects of the central and peripheral
modes of action of acupuncture as they impact on the
hypothalamic-pituitary-ovarian axis and the pelvic organs,
respectively. Moreover, the effect of acupuncture on anxiety and
stress and ensuing potential indirect effects on female
fertility will also be discussed.
Background
Acupuncture is the manipulation of thin metallic needles
inserted into anatomically defined locations on the body to
affect bodily function. The US Food and Drug Administration has
recently removed acupuncture needles from the category of
experimental medical devices and now regulates them just like it
does other devices, such as surgical scalpels and hypodermic
needles, under good manufacturing practices and single-use
standard of sterility (1).
The general theory of acupuncture is based on the premise that
there are patterns of energy flow (Qi) through the body, which
are essential for health. Disruption of this flow is believed to
be responsible for disease. Acupuncture can correct imbalances
of flow at identifiable points close to the skin.
According to the proposed international acupuncture nomenclature
by The World Health Organization in 1991 (17), the meridian
system consists of 20 meridians interconnecting about 400
acupoints. These acupoints correspond to specific areas on the
surface of the body, which demonstrate higher electrical
conductance because of the presence of higher density of gap
junctions along cell borders. They act as converging points (or
sinks) for electromagnetic fields. A higher metabolic rate,
temperature, and calcium ion concentration, are also observed at
these points. In principle, positive (anode) pulse stimulation
of a point inhibits the organ function, whereas negative
(cathode) pulse stimulation enhances that function (18). This
forms the basis of electroacupuncture, which applies small
electrical needles inserted in specific acupoints.
Effects of acupuncture on the hypothalamic-pituitary-ovarian
axis and menstrual cycle
Although traditional Chinese medicine understanding of
acupuncture is based on ancient medical theory, a modern and
scientific neuroendocrine perspective has begun to evolve in the
past two decades. Mayer et al. (19) first reported that
acupuncture analgesia was induced through endorphin production
and antagonized by the narcotic antagonist naloxone. Other
studies similarly suggested that certain effects of acupuncture
are mediated through the nervous system, within which
ß-endorphin and other neuropeptides have been implicated
(20-22).
Acupuncture was shown by Petti et al. (20) to cause a
significant increase in ß-endorphin levels during treatment,
which lasted for up to 24 hours. ß-endorphin is derived from its
precursor protein pro-opiomelanocortin, which is present in
abundant amounts in neuronal cells of the arcuate nucleus of the
hypothalamus, pituitary, medulla, and in peripheral tissues
including intestines and ovaries (23-25). Pro-opiomelanocortin
cleaves to form adrenocorticotropic hormone and ß-lipoprotein.
Further cleavage of ß-lipoprotein yields neuropeptides including
ß-endorphin. Aleem et al. (26, 27) demonstrated the presence of
immunoreactive ß-endorphin in follicular fluids of both normal
and polycystic ovaries.
The influence on gonadotropin secretion and the menstrual cycle
by endogenous opioid peptides is believed to be mediated by
their action on GnRH secretion (28). The hypothalamic
ß-endorphin center and the GnRH pulse generator, in fact, are
both situated within the arcuate nucleus. Quigley et al. (29)
first reported an increased opioid inhibition of LH secretion in
hyperprolactinemic patients with pituitary microadenomas. Ching
(30) and Orstead and Spics (31), respectively, showed that
opioid peptides suppress GnRH release in rats and rabbits.
The role of these neuropeptides, including ß-endorphin, in the
regulation of GnRH secretion in humans has recently been
reviewed by Kalra et al. (32) and Pau and Spies (33). Rossmanith
et al. (34) demonstrated the role of opioid peptides in the
initiation of the mid-cycle LH surge in normal cycling women.
Meanwhile, measurement of ß-endorphin in ovarian follicular
fluid of healthy ovulatory women revealed much higher levels
than that in circulating plasma (35). The highest level of
ß-endorphin was noted to be in the preovulatory follicle.
Because acupuncture treatment impacts on ß-endorphin levels,
which in turn affect GnRH secretion and the menstrual cycle, it
is logical to hypothesize that acupuncture may influence
ovulation and fertility. Animal studies have revealed that
acupuncture treatment normalized GnRH secretion and affected
peripheral gonadotropin levels (36, 37). Various investigators
have shown that in normally ovulatory or anovulatory women,
acupuncture also influenced plasma levels of FSH, LH, E2, and P
(38-40). Acupuncture as a surrogate for hCG in ovulation
induction was successfully used by Cai (41). Chen and Yu (42)
showed that electroacupuncture normalized they
hypothalamic-pituitary-ovarian axis, and in another study Chen
(43) reported that 6 of 13 anovulatory cycles responded to
acupuncture treatment.
A series published from the University of Heidelberg in Germany
(44) used auricular acupuncture on 45 infertile women suffering
from ovulatory dysfunction such as oligomenorrhea and luteal
phase defect. The control group received medical treatment
including bromocriptine, dexamethasone, levothyroxine,
clomiphene citrate (CC), and gonadotropin. Although the
investigators concluded that resumption of ovulatory cycles
occurred significantly more often in the acupuncture group
compared to the control group, pregnancy rates were not
different between the two groups. However, interpretation of
study data was very difficult due to the heterogeneity of the
patient population and treatment modalities. Moreover, seven
pregnancies in the acupuncture group were actually achieved with
hormone treatment 6 months after acupuncture was stopped.
Another study by Stenver-Victorin et al. (45) evaluated the use
of electroacupuncture for ovulation induction on 24 oligo/amenorrheic
women with polycycstic ovarian syndrome (PCOS). The percentage
of ovulatory cycles in all subjects was shown to improve from
15% (in a total of 3 months before treatment) to 66% up to 3
months after treatment. Responsive patients were noted to have
significantly lower body mass index (BMI), waist-to-hip
circumference ratio, serum T concentration, serum T/sex
hormone-binding globulin ratio, and serum basal insulin level.
They suggested that, in these selected patients with PCOS,
acupuncture could be considered as an alternative or adjunct to
pharmacological ovulation induction.
A recent prospective randomized controlled study by Paulus et
al. (46) compared pregnancy rates in a total of 160 patients
undergoing IVG. Acupuncture was performed in 80 patients 25
minutes before and after ET. After controlling confounding
variables, clinical pregnancy rate for the acupuncture group
(42.5%) was significantly higher than the control group (26.3%).
Peripheral effects of acupuncture
In addition to the central modulation of the
hypothalamic-pituitary-ovarian axis, the effects of acupuncture
on the autonomic nervous system have been well documented (47).
In the early 1980s, Yao et al. (48) reported long-lasting
cardiovascular depression induced by acupuncture stimulation of
the sciatic nerve in unanesthetized hypertensive rats. In the
human, acupuncture was also shown to be sympathoinhibitory.
After acupuncture, sympathetic nerve activity as measured by
norepinephrine level, skin temperature, blood pressure, and pain
tolerance threshold was shown to be decreased (49).
Endometrial thickness, morphology, and uterine artery blood flow
have been implicated as important parameters for success of
implantation of human embryos (50-57). Despite conflicting
results in the utilization of these parameters during various
stages of treatment to predict outcome in IVF, it is generally
believed that adequate endometrial thickness is required to
optimize pregnancy rate. Because endometrial thickness is a
function of uterine artery blood flow, Sher and Fisch (58)
reported a novel method of using vaginal sildenafil in an
attempt to improve uterine artery blood flow and endometrial
development in patients undergoing IVF.
With its central sympathoinhibitory effect, acupuncture may
contribute to reduce uterine artery impedance and therefore,
increase blood flow to the uterus. In fact, Sterner-Victorin et
al. (59) demonstrated this when they performed acupuncture in 10
infertile women who were down-regulated by GnRH analog to avoid
the effect of endogenous hormone on the uterine artery blood
flow.
Pulsatility index in the uterine artery and skin temperature (on
the forehead and lumbosacral area) were evaluated in three time
periods-before, right after, and 2 weeks after acupuncture
treatment (twice a week for 4 weeks). Pulsatility index and skin
temperatures were found to be significantly decreased and
increased, respectively, both right after and 14 days after
acupuncture treatment. This effect was hypothesized to be caused
by central inhibition of sympathetic activity.
Acupuncture and stress reduction
It has been well documented that infertility causes stress
(60-65), and stress reduction may, in turn, improve fertility
(66). However, the relationship between stress and infertility
is that of a vicious cycle. Social stigmatization, decreased
self-esteem, unmet reproductive potential of sexual
relationship, physical and mental burden of treatment, and the
lack of control on treatment outcome are just some of the
factors that can lead to psychological stress in any couple
pursuing infertility treatment. In turn, stress may lead to the
release of stress hormones and influence mechanisms responsible
for a normal ovulatory menstrual cycle through its impact on the
hypothalamic-pituitary-ovarian axis.
The use of acupuncture for reducing anxiety and stress possibly
through its sympathoinhibitory property and impact on
ß-endorphin levels has been reviewed (67, 68), and the efficacy
of acupuncture in depression has also been studied (69). Because
the pharmacological side effects of anxiolytic and
antidepressant drugs on infertility treatment outcome are
largely unknown, acupuncture may provide an excellent
alternative for stress reduction in women undergoing infertility
treatment.
Discussion
The practice of acupuncture to treat identifiable patho-physiological
conditions has been a subject of intense research. The
underlying physiologic mechanisms of acupuncture such as the
release of opioids and other peptides in the central peripheral
nervous system, and its inhibition of the sympathetic nervous
system have been increasingly established. Promising results
from credible trials have emerged for the use of acupuncture in
treating various pain syndromes, substance abuse, and
chemotherapy-induced nausea and vomiting.
Although the definitive role of acupuncture in the treatment of
female infertility is yet to be established, its neuroendocrine
effect on the hypothalamic-pituitary-ovarian axis and the
preliminary clinical data reviewed here justifies further
clinical trials to systematically examine the efficacy of
acupuncture in treating various conditions related to female
infertility such as ovulatory dysfunction associated with PCOS.
The peripheral impact of acupuncture in improving uterine artery
blood flow and hence endometrial thickness also provides
encouraging data regarding its potential positive effect on
implantation.
Whether these potential beneficial effects of acupuncture on the
reproductive system can be translated into improving infertility
treatment outcomes will eventually mandate randomized controlled
studies of adequate design. Because acupuncture is nontoxic and
relatively affordable, its indications as an adjunct in assisted
reproduction or as an alternative for women who are intolerant,
ineligible, or contraindicated for conventional hormone
induction of ovulation deserves serious research and
exploration.
Appropriate training, credentialing, and certification of
acupuncture practitioners by state agencies can facilitate the
integration of acupuncture into the treatment of female
infertility, and healthcare in general. The NIH Consensus
Conference (1) agreed that this is necessary to allow the public
and other health practitioners to identify qualified acupuncture
practitioners. With the help of the US Department of Education,
issues of training and licensure of non-physician and physician
practitioners have been addressed. There is sufficient evidence
to acupuncture's value to expand its use into conventional
medicine and treatment of female infertility, and to encourage
further studies of its underlying mechanisms as well as to
establish its clinical value.
Acupuncture Normalizes Dysfunction of
Hypothalamic-Pituitary-Ovarian Axis
By Bo-Ying
Chen M.D., Professor of Neurobiology
Institute of Acupuncture and Department of Neurobiology
Shanghai Medical University, Shanghai 200032, P.R. China
(Received June 3, 1997; Accepted with revisions June 30,1997)
ACUPUNCTURE & ELECTRO-THERAPEUTICS RES., INT. J., Vol. 22, pp.
97-108, 1997
Copyright (c) 1997 Cognizant Communication Corp. Printed in the
USA. 0360-1293/95 $10.00 + .00
ABSTRACT This article summarizes the studies of the
mechanism of electroacupuncture (EA) in the regulation of the
abnormal function of hypothalamic pituitary-ovarian axis (HPOA)
in our laboratory. Clinical observation showed that EA with the
effective acupoints could cure some anovulatory patients in a
highly effective rate and the experimental results suggested
that EA might regulate the dysfunction of HPOA in several ways,
which rneans EA could influence some gene expression of brain,
thereby, normalizing secretion of some hormones, such as GnRH,
LH and E2. The effects of EA might possess a relative
specificity on acupoints.
KEY WORDS: Electroacupuncture, ß-Endorphin, GnRH, LH,
Estradiol, Estrogen receptor, Ovariectomized rat,
Hypothalamic-pituitary-ovarian axis
INTRODUCTON
Acupuncture
is a treasure of Chinese traditional medicine, which is employed
in the treatment of different diseases, especially in relief of
all kinds of pain [1, 2] over the world. Since 1960s we have
used acupuncture with appropriate electro-stimulation to cure
patients with anovulation disorder (sterility), the rate of EA
induction of ovulation was increased from 50% initially to 80%
presently. Other authors in China also reported that acupuncture
was successfully to treat patients with sterility [3] and the
lying-in woman with subnormal contraction of uterus [4]. All the
above research demonstrates that acupuncture may be an effective
curative method of some woman's diseases. However, many
questions, such as "why", "how to" and "which" about the
mechanism of EA effect are unknown. To address these problems we
supposed that EA might influence the production and secretion of
hormones, neurotransmitters or neuro-modulators of HPOA leading
to the normalization of hormone status. We also noticed certain
artides reported that EA might affect the blood levels of LH,
FSH, estradiol (E2) and prolactin in the female patients [4, 5,
6] and EA may be related to long term changes in gene expression
[7, 8]. These results are all significant, yet insufficient to
explain the mechanism of EA in the regulation of the function of
HPOA. To obtain more data, a series of experimental studies in
human and animal models has been performed in our laboratory.
MATERIALS AND METHODS
Selection and treatment of cases
Ten cases of chronically anovulatatory patients including eight
cases of polycystic ovarian disease (POCA), one case of
hypogonadotropic amenorrhoea and one case of oligomenorrhea were
treated with EA in 13 menstruation cycles. They were all of
productive age and the courses of disease were 3 to 12 years. On
the 10th day of each menstruation cycle, the patients accepted
the EA treatment. "Guanyuan(RN4)," "Zhongji(RN3)," "Sanyinjiao(SP6),"
and bilateral "Zigong(EXCA1)" points were stimulated for 30 min
at 8:00 AM, Q.D. for 3 days. The stimulation parameters were
7-8mA and 4-5 Hz with G6805 model generator. The electric
current of EA was bearable well for every patient. The blood
samples were collected from forearm of the patients one time per
15 min for detection of FSH.LH and ß-endorphin (ß-E).
Five health volunteers of a productive age with normal
menstruation cycle were selected as controls, which were
undergone the same treatment as above mentioned.
Animals and treatments
Wistar female rats weighting 200-250g were used. The half of
animals were undergone ovariectomy and fed in the same
environment with the intact rats at least for 15 days and
vaginal smears were examined per day for 3 times. No exfoliative
epithelium cell was found in the smears as an index for
successfill ovariectomy. The ovariectomized rats and intact rats
were randomly divided into two groups respectively:
ovariectomized rat group (OVX), ovariectomized rat accepted EA
treatment group (OVX+EA), intact rat group (INT) and intact rat
accepted EA treatment group (INT+EA). The animals in OVX+EA and
INT+EA received EA at the experimental acupoints of Guanyuan
(RN4), Zhongji (RN3), Sanyinjiao (SP6) and bilateral Zigong
(EXCA1) by EA apparatus (Model G6805-2, SMIF, Shanghai, China)
with the frequency of 3 Hz and an intensity to produce a slight
twitch of the limbs. After 3 days' treatment animals were given
EA at Waiguan (SJ5) and Huatuojiaji (EXTRA21) as the control
acupoints in the same way (Fig 1). By the end of last
experiment, animals were sacrificed and their adrenals, brains
and pituitaries were taken out for detection of nucleolar
oganizer regions (AgNORs) and hormones.
Pushpull perfusion in hypothalamic preoptic area (POA) and
elution of pituitary and LH and ß-endorphin (ß-EP)
The technique of brain pushpull perfusion was processed as
previously described by our laboratory [1]. The perfusate from
hypothalamic POA was kept at -70°C for GnRX and ß-EP RIA.
The pituitaries were retrieved and put into 4°C cooled saline.
Afterward, each pituitary was homogenized with 500µl of 70%
acetone aqueous solution at 4°C. The homogenate was
centrifugalized (2,000xg for 15 min at 4°C) and the supernatant
was freeze-dried for LH and ß-EP RIA.
Radioimmunoassay (RIA) of hormones
GnRH IRA: GnRH content in the perfusate from rat
hypothalamus was determined by RIA method developed by Nett in
1973 [9]. GnRH was iodinated by the modified chlomine-T
technique[10]. Na125
I was manufactured by Radiochemical
Center, Amersham.
ß-EP RIA: The sensitive radioimmunoassay was a routine in
our laboratory [1]. The standards of human and rat ß-EP was
synthesized by Peninsula Laboratories, Inc. and the rabbit
antiserum of both ß-EP was developed in our laboratory. The
cross-reaction from human ß-EP and camel ß-EP was detected about
20%. The sensitivity of this method was 10pg/tube.
LH, E2 and
corticosterone RIA: LH, E2
and corticosterone RIA kits were bought from Shanghai Institute
of Biologic Products, the Ministry of Health, P.R. China. All
procedures of RIA were performed as described in the kit
manuals.
|
Fig.
1 |
A: |
Sketch of ventral view (left) and dorsal view (right) of
rat shows the acupoints we used |
|
B: |
Diagram shows the electroacupuncture procedures in
conscious rat |
Staining
techniques:
Vaginal smears were fixed by 100% ethyl alcohol, then stained
with HE method. Adrenal sections were cut in 4µm thickness from
paraffin blocks and processed with silver nitrate staining
technique[11]. In each case, one hundred cells in zona fascicula
were examined randomly under 100-fold oil immersion lens.
Numbers and sizes of AgNOR dots were counted and measured.
C-fos protein immunohistochemistry: The
inmunohistochemical analysis of c-fos expression in rat brain
was perforrned as previously described[11].
Estrogen receptor (ER) protein immunohistochemistry (ABC
method): Under sodium pentobarbital anesthesia (50 mg/kg, ip),
the animals were perfused via left cardiac ventricle with 100ml
of phosphate-buffered saline (PBS), followed by 300ml ice-cold
fixative containing 4% paraformaldehyde in 0.1 M phosphate
buffer (pH7.4). Afterwards, brain was removed with the same
fixative for one day and immersed in 0. lM phosphate buffer
containing 30% sucrose for another day. The hypothalamus blocks
were frozen with dry ice and cut into 35 µM thick section by
cryostat. The brain sections were washed with 0.01M PBS for
15min x 3 and incubated in 0.01M PBS containing 0.5% Triton 100
and 3% normal goat serum (NGS) at 37°C-for one hour. Afterwards,
the sections incubated in 1:1,000 ER monoclonal antibody (H222,
Abott Co.) at 37°C for one hour, then at 4°C for two days. The
sections, washed in PBS three times, were processed by ABC kit
(from Vecot Labs) induding sequential incubation at 20°C in the
following solutions with washes between them. (1). second
antibody (dilution 1:100), 30min. (2). A+B reagents
(dilutionl:100), 60min. (3). 0.05% diaminobenzidine/ 0.02%
hydrogen peroxide in 0.1M Tris- HCI buffer (pH 7.2) 10min. The
sections were washed in tap water, mounted and examined under
light microscope. The certain areas of typical immunoreactive
positive neurons were measured by computer image analysis system
(Vecta PC).
ER mRNA hybridization: The total mRNA of brain was eluted
by the modified phenol method [12]. ER cDNA probe (244bp) was
labeled by the DlG-labeling kit (from Bohringman Co., Germany).
The dot blot hybridization was processed as the method described
by Sambrook J and his colleagues [13]. The dot blot images were
analyzed with gray density by computer imaging analysis software
(TJTY-300, from Tong -Ji university, Shanghai, China).
Statistics: All data in this paper were treated with
analysis of variation (ANOVA), least significant difference
(ISD) or student T-test.
RESULTS
Effect of EA on ovulatary induction and curing sterility in
woman
After EA the blood ß-EP level of the patients resulting in
ovulation either declined or maintain at the levels within the
range of the normal levels and the ß-EP levels of those failing
to show ovulation were significantly higher than the normal's'
(table 1). On the other hand, the blood LH and FSH levels of the
patients with ovulation after EA treatment tended to be the
normal [14].
Table 1. Change of blood ß-EP level
before and after EA (pg/ml)
|
Group of cases |
N |
Before EA |
After EA |
|
|
|
Ovulation |
6 |
65.59 ± 24.15 |
*38.86 ± 10.11 |
|
No ovulation |
7 |
65.59 ± 24.15 |
80.09 ± 22.16 |
|
Control |
5 |
38.84 ± 10.13 |
41.52 ± 6.40 |
The values in this table are mean±SE, *P<0.05
Effect of EA on dysfunction of HPOA in ovariectomized rats
For a further study of the mechanism of EA effect on HPOA a
series of experiments in the animal models was performed.
(1). EA induces maturation and exfoliation of vaginal
epithelium cell and enhances blood level of E2.
After
ovariectomy two weeks late, the exfoliated epithelium cell
disappeared from the vaginal smears of the rats, but it
reappeared in the smears following EA treatment. The blood level
of E2 in OVX was increased significantly (table 2). No obvious
change was seen in INT after EA treatment and in OVX following
EA treatment with the control acupoints.
Table 2. The level of blood E2
following EA treatment (pg/ml)
|
Group |
N |
Before EA |
After EA |
|
|
|
OVX |
10 |
*5.47 ± 0.63 |
**11.58 ± 0.98 |
|
INT |
10 |
18.00 ± 3.26 |
18.34 ± 8.77 |
*P < 0.05 compared with INT, **P<0.01 compared with before EA
(2). EA promotes enlargement of adrenals and enhances
activity of adrenal AgNORs as well as blood level of
corticosterone
We found the adrenals of OVX+EA were enlarged and the weight of
the adrenals was raised significantly. Using histochemical
method, the AgNORs of the cells in inner adrenal cortex were
examined. The result shows that the activity of AgNORs of OVX
was enhanced (table 3, 4), and the level of blood corticosterone
in OVX+EA was also increased (table 5). There were no similar
effects in INT following EA treatment and in OVX after EA with
control acupoints.
Table 3. AgNORs number in OVX and INT
Group
N |
INT
4 |
INI+EA
3 |
OVX
4 |
OVX+EA
7 |
F
value |
|
|
Number
of AgNORs
(mean/100 cells) |
1.55
1.82
1.24
1.30 |
1.19
1.28
1.16 |
1.25
1.61
1.66
1.96 |
2.53
2.05
1.82
2.86
2.86
2.93
3.92 |
9.614* |
|
|
*P < 0.01
tested with ANOVA
Table 4. Weight of adrenal
Group
N |
INT
5 |
INI+EA
3 |
OVX
5 |
OVX+EA
8 |
F
value |
|
|
Weight
(mg) |
57
56
57
43
57 |
54
57
58 |
45
68
56
50
58 |
67
72
66
71
57
74
74
68 |
5.825* |
|
|
*P < 0.01
tested with ANOVA
Table 5. The levels of blood
corticosterone in OVX and lNT (mean ± SE, ng/ml)
|
Group |
N |
Before EA |
After EA |
|
|
|
OVX |
12 |
4.78 ± 0.42 |
*6.06 ± 0.73 |
|
INT |
12 |
3.64 ± 0.15 |
4.76 ± 1.25 |
*P < 0.001 compared with before EA
(3). EA decreases the level of hypothalamic GnRH, pituitary
LH and increases the contents of hypothalamic and pituitary
ß-endorphin
After EA treatment the levels of GnRH released from hypothalamus
was rnarkedly decreased however, the ß-endorphin (ß-EP)
secretion in hypothalamus was raised. The pituitary content of
LH was also fallen, but the ß-EP of pituitary was increased, as
well as peripheral LH and ß-EP level (Fig.2).
|
Fig. 2 |
Change of hypothalarnic GnRH and ß-EP, pituitary LH and
ß-EP, blood LH and ß-EP before and after EA |
Effect of EA on brain c-fos expression in ovariectomized rats
The area occupied by FOS protein labeled neuron was detected in
medial preoptic nucleus (MPN), lateral preoptic nucleus (LPN),
suprachiasmatic nucleus (SCN), paraventricular nucleus of the
hypothalamus (PAVN), medial amygdala nucleus (MAN),
periventricular nucleus of the hypothaLsmus (PVN), ventromedial
nucleus of the hypothalamus (VNH) and arcuate nucleus (AR) 4
hours after ovariectomy (fig. 3a). The C-fos immunoreactive
labeled neurons disappeared two weeks later following
ovariectomy. The rats recovering for more than two weeks after
ovariectomy, were received EA treatment. Many specific FOS
labeled cells were observed in LPN, VNH, SCN and especially in
POA, ARN, and PVN, but not any labeled neuron could be found in
MAN. No obvious C-fos expression was shown in those nuclei in
INT and INT+EA (fig. 3b).
|
Fig. 3a |
C-fos immunocytochemistry neurons distribution after
ovariectomy |
|
Fig. 3b |
C-fos expression labeled neurons following
electroacupuncture |
Effect of EA on expression of ER protein and ER
mRNA in rat brain
Estrogen receptor (ER) immunoreactive neurons were observed
widely in rat brain with immunohistochemical technique,
especially in MPN, ARN and VNH. The above nuclei were measured
by computer image analysis system, and the results show that the
mean gray density in OVX+EA was decreased apparently compared
with that in OVX. Whereas there were no obvious changes of gray
density levels in INT and INT+EA (fig, 4).
|
Fig. 4 |
Effect of EA on expression of ER protein in rat brain (Immunohistochernistry
of monoclonal antibody) *p < 0.01 compared with OVX |
The dot blot indicated that ER mRNA expression was
increased about 48.11% in OVX compared with INT. The gray
density of OVX was 129.75 ± l2.l3 and that in OVX+EA was 199.25
± 5.75 attenuated significantly (Fig. 5). The gray density level
in INT was 87.60 ± 5.91, and the level in INT+EA was 83.60 ±
4.83. There was no significant difference between INT and INT+EA
|
Fig. 5 |
Effect of EA on expression of ER mRNA in rat brain (dot
blot) *** p < 0.01 compared with OVX
|
DlSCUSSION
Since 1985 we have observed that the effect of EA ovulatary
induction might relate to the hand skin temperature (HST) and
the blood level of ß-EP [14]. On the other hand, after EA the
blood FSH and LH levels of the patients who successfully
ovulated either declined or maintained at normal. In general,
provided that body temperature was normal and the environmental
temperature was constant round 25°C, the HST may reflect the
state of sympathetic system of a patient. These results suggest
that in anovulatary cases the hyperactive sympathetic system can
be depressed by EA and the function of HPOA can be regulated by
EA through central sympathetic system. Moreover, EA may mediate
the abnormal function via the influence on the secretion of the
hormones in the different Level of HPOA.
To gain more evidences, we designed some animal experiments to
explain the mechanism of EA effects on HPOA at the whole,
cellular and molecular levels. We found that EA can induce
maturation and exfoliation of vaginal epithelium cell in OVX
rat. It is known that maturation and exfoliation of vaginal
epithelium cells are a reaction dependent on estrogen level. So
we determined the level of blood E2 in OVX and OVX+EA. The
result shows the level of blood E2 in OVX was lower than that in
normal, but it was increased significantly after OVX accepted EA
treatment with the experimental acupoints. This result suggests
EA might promote the activity of the compensative mechanism to
elevate the subnormal level of E2 induced by ovariectomy in
rats.
What is this compensative mechanism? To resolve this question,
we considered that adrenal is the main organ to secrete sexual
hormones except ovarian in females and observed the adrenals of
the animals in four groups. The results show that the mean
weight of the adrenal in OVX+EA was higher than that in OVX, INT
and INT+EA, suggesting the adrenal function might be activated
by EA. Subsequently, we detected that the number of AgNORs in
zona fasciculata of OVX+EA was significantly increased.
Nucleolar organizer regions (NORs) are loops of DNA, which
possess ribosomal RNA (rRNA) genes. They are of vital
significance in the ultimate synthesis of protein. Thus, the
number and configuration of AgNORs (NORs stained by silver
staining method) may reflect the activity of cell
differentiation and transcription of nucleolar rDNA [15]. In the
same time we found the content of blood corticosterone in OVX+EA
was raised markedly, but there was no change of blood
corticosterone in OVX, INT and INT+EA. This result provided a
further evidence that the adrenal cortex cells were initiated in
OVX+EA.
The results including the changes of GnRH releasing from
hypothalamus and of the pituitary and blood LH contents suggest
that the effects of acupuncture in the regulation of HPOA may be
exerted via to promote the function of hypothalamic
pituitary-adrenal axis (HPAA), increasing the synthesis and
secretion of adrenal steroid horrnones, the androgen of which
then be transformed into estrogen in other tissues and thereby
reset the negative feedback of estrogen to HPOA. Moreover, EA
may accelerate the release of brain and pituitary ß-EP to
inhibit the overnormal secretion of GnRH and LH that may be
normalized.
Recently immunohistochemical analysis of the expression of
oncogene c-fos ABl was induced by variety of stimuli [16, 17].
This represents a new method for mapping neuronal activity at
the cellular level [18] and thus functionally and systematically
tracing neuronal pathway in the nervous system (C NS) [19]. We
used this method to examine the distribution of FOS labeled
neuron in CNS for recovery of more evidences that EA may alter
the neuroendocrine function of HPOA in ovariectomized rats in
cellular and gene level. The results show that the specific FOS
labeled neurons were observed especially in POA, ARN and PVN in
OVX following EA treatment. In above nuclei there were a high
concentration of GnRH and ß-EP neuron [20]. These results
suggest this fact that the expression of FOS labeled neurons
reappeared in above mentioned areas following EA treatment in
ovariectomized rats may be related to the changes of GnRH and
ß-EP from rat hypothalamus after EA treatment.
The level of estrogen in the body may regulate the expression of
ER, which may by down-regulated following increase of estrogen
level and up-regulated after decrease of estrogen [22]. Our
finding that after decline of blood E2 induced by ovariectomy
the expression of ER was increased and the expression of ER was
inhibited by EA inducing the elevation of blood E2 are in
accordance with these reported results. ER existing in the
brain, especially in POA, ARN and VHN may mediate the function
of neuroendocrine system [22, 23]. Thus, our observations
suggest that the influence of EA on the change of ER expression
in brain may be one of further mechanisms of EA normalizing the
dysfunction of HPOA.
INT rats as experimental control we adopted were all of in the
stage of preestrus and estrus because the animal sexual hormes
and brain ER expressions were changed with the sexual cycle
[24]. All INT rats were selected to fix in the two stages there
may be a relative constant comparability.
Our results show no same effects were seen after EA treatment in
INT and following EA with control acupoints in OVX, suggesting
that EA may possess a relative specificity on acupoint and the
effect of EA may be a kind of normalization.
CONCLUSION
Our observations reveal that acupuncture may regulate the
abnormal function of HPOA in many ways, which means that
acupuncture may activate C-fos expression of brain, then a long
term changes at molecular level would start, following the
regulation of gene expression in FOS relative gene, such as ER
mRNA and GnRH mRNA involved. On the other hand, EA may promote
the activity of the body compensative mechanisms, then the
levels of hormones, such as GnRH, LH, estrogen and so on would
be normalized. The effect of acupuncture on regulating the
function of HPOA may possess a relative specificity of acupoint.
Moreover, our clinical and animal experimental results suggest
that it is necessary for obtaining a satisfactory effect that
proper stimulation should be about thirty minutes Q.D. for three
days. This suggestion provides a successful consideration for
clinical practice in curing the woman patients with dysfunction
of sexual endocrine, such as primary ovarian dysfunction,
climacteric syndrom, after-ovariectomy and polycystic ovarian
disease etc.
ACKNOWLEDGMENT
The work was supported by National Natural Foundation of China
(3880910 and 392708340) and a grant from the State Key
Laboratory of Medical Neurobiology of China (92003).
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