Interleukin-6 and Hepcidin Levels during Hormone-Deplete and Hormone-Replete Phases of an Oral Contraceptive Cycle: A Pilot Study.

BACKGROUND: In the past, elevated estradiol levels were reported to downregulate the iron regulatory hormone hepcidin, thereby potentially improving iron metabolism. As estrogen plays a role in regulating the menstrual cycle and can influence the cytokine interleukin-6 (IL-6; a hepcidin up-regulator), this investigation examined the effects of estradiol supplementation achieved by the use of a monophasic oral contraceptive pill (OCP) on IL-6, hepcidin levels and iron status during the hormone-deplete versus hormone-replete phases within an oral contraceptive cycle (OCC). METHODS: Fifteen healthy female OCP users were recruited and provided a venous blood sample on 2 separate mornings during a 28-day period. These included (a) days 2-4 of the OCC, representing a hormone-free withdrawal period (WD); (b) days 12-14 of the OCC, representing the end of the first week of active hormone therapy (AHT). RESULTS: IL-6 and hepcidin levels were not significantly different at WD and AHT. Serum ferritin was significantly higher (p = 0.039) during AHT as compared to WD. CONCLUSIONS: Fluctuations in OCP hormones (estradiol and/or progestogen) had no effect on basal IL-6 and hepcidin levels in young women. Nevertheless, elevated ferritin levels recorded during AHT may indicate that OCP hormones can positively influence iron stores within an OCC despite unchanged hepcidin levels.

The anti-hyperplasia of mammary gland effect of protein extract HSS from Tegillarca granosa.

Tegillarca granosa Linnaeus, possesses various biological functions and has been used a Chinese traditional medicine more than one century, but there is no report about anti-hyperplasia of mammary gland (HMG) activity of drugs from T. granosa. In this study, we investigated the anti-HMG effect of protein extract named HSS from T. granosa. The HMG model of virgin female Sprague Dawley rats was prepared by injecting estrogen in the thigh muscle of the rats and progestogen consecutively. HMG rats were treated with either HSS or positive control drug by i.g. for 35 consecutive days. In order to evaluate anti-HMG activity of HSS, Changes of nipple height and diameter, serum sex hormones levels, organ indexes and pathologic changes of mammary gland were performed. Body weight, food intake, pathomorphology examination of organs (heart, liver, spleen, lung, kidney), hematological and biochemical analysis were performed to evaluate the toxicity of HSS. HSS could significantly reduce nipples height and diameter, increase P concentration of HMG rat serum, spleen and thymus index, decrease uterus index, and has therapeutic effect on rat HMG and no toxicity at 500mg/kg/day. The anti-HMG mechanism of HSS may be related to AP-2α and P53. HSS has protective and therapeutic effects on HMG rats, and may be a promising agent for treating hyperplasia of mammary glands.

[Study on preparation method of Yueanjian for treatmen of premenstrual syndrome].

OBJECTIVE: To establish the liver-depression and spleen-deficiency syndrome model in rats to screen the optimal extraction method of small compound Yueanjian on the basis of pharmacodynamic and chemical indicators. METHOD: The PMS liver-depression and spleen-deficiency syndrome model were established by the chronic restraint stress method and treatment with Yueanjian extracted by three methods: water-extraction, steam-distillation and alcohol-extraction. Behavioral performances and the contents of estradiol and progestin in serum were determined before and after the administration of the three extracts. The contents of salvianolic acid B in these three extracts were detected by HPLC. The optimal extraction method of Yueanjian was selected according to pharmacodynamic results. RESULT: The contents of estradiol and progestin in groups treated with steam distillations and alcoholic extraction were higher than the model group. In the open field test, the group treated with steam distillations showed much higher scores than the model group. HPLC showed that the content of salvianolic acid B extracted by steam-distillation was higher than the other two extracts. CONCLUSION: On the basis of pharmacodynamic and chemical results, the steam-distillation was proved to be best extraction method of Yueanjian.

Distinct gender-related sleep pattern in an acute model of TMJ pain.

Since it is recognized that acute inflammation of the temporomandibular joint results in sleep disturbances in male rats, and that the orofacial region may display a site-specific effect of ovarian hormones on nociception, we hypothesized that distinct genders would respond differently when subjected to this inflammatory acute orofacial pain. Sleep was monitored after injection of saline/Freund's adjuvant into the temporomandibular joint in male and female (proestrus and diestrus phases) rats. Progesterone and stress-related hormones were also assessed. In males, Freund's adjuvant induced a significant nociceptive response and sleep disturbances. Behavior and sleep architecture in the females remained unaffected. Our results suggest that females and males present distinct responses to an acute model of orofacial pain.

[Experimental study on Ruxian Pill I in treating mammary gland hyperplasia rabbits].

OBJECTIVE: To observe the effect of Ruxian Pill I (RXP I) on mammary gland hyperplasia (MGH) in rabbits and to explore its mechanism. METHODS: Sixty rabbits were divided into the high, medium and low dose of RXP I groups, the Xiaoyao Pill (XYP) group, the model group and the normal control group with 10 in each group. The former 5 groups were injected with diethylstilbestrol and progesterone intramuscularly for one month to induce the MGH model and then given respective medicines via gastrogavage for 3 months. The changes in morphology of mammary gland were observed using light and electronic microscope, the levels of estradiol (E2) and progestogen (P) were measured by radioimmunoassay, and the expression of estradiol receptor (ER) and progestogen receptor (PR) were detected with immunohistochemistry before, at the end of and 3 months after the treatment. RESULTS: Compared with those before treatment and those in the model group, in the high and midium dose of RXP I groups after treatment, obvious decrease of acini number in hyper-plastic lobuli mammae, connective tissues and blood capillaries, layers of glandular epithelium cells and organellers were seen with partial of hyperplastic cell apoptosis in them. Besides, the serum E2 level decreased obviously (P < 0.05), while the serum P level increased, and the ER expression down-regulated significantly (P < 0.05), but no obvious changes of PR expression was found. Three months later, all the above indexes maintained stable without rebound. CONCLUSION: RXP I treatment could alleviate the hyperplasia of mammary glands, reduce E2 level, and down-regulate ER expression in rabbits with MGH, showing a significant therapeutical effect.

Progesterone bioavailability with a progesterone-releasing silicone vaginal ring in IVF candidates.

A vaginal ring made of silicone polymers and barium sulfate, and containing 1 g of pure micronized progesterone, was developed for luteal supplementation in women undergoing cycles of in vitro fertilization (IVF). The ring, modeled on the Estring, was designed as a means of providing continuous intravaginal delivery of progesterone. Bioavailability of progesterone in the blood was demonstrated for 24 hours in IVF candidates who had an endogenous progesterone deficiency after treatment with gonadotropin-releasing hormone (GnRH) analogues. After the first 4 h of increasing release of progesterone from the ring (with mean serum levels of 1.39 +/- 0.8 ng/ml after 4 h), only a slight increase in serum progesterone levels (with a mean peak of 1.5 +/- 0.45 ng/ml after 24 h) was observed during the rest of the test period. Gonadotropin levels were not affected after insertion of the ring. The ring was well tolerated by the patients. The maximum serum progesterone level was lower in comparison with other forms of progesterone application, but it should be sufficiently high, due to the uterine first-pass effect. This study demonstrated that progesterone administration through a silicone ring for luteal support is feasible in IVF treatment. As the vaginal ring is very well tolerated by the patients, these findings may encourage the pharmaceutical industry to design an appropriate progesterone ring for luteal support.

Dynamics of progestogenic activity in serum following administration of Ligusticum chuanxiong.

Many women are using botanical alternatives for menopausal hormone replacement therapy (HRT) because current progestins, compounds with progesterone activity, have adverse risk profiles. However the development of phyto-progestins for HRT is hampered by the absence of basic pharmacokinetic/pharmacodynamic (PK/PD) data due to the lack of methods to capture summated effects of the numerous compounds that contribute to bioactivity in vivo. In this study, we explored the utility of progesterone receptor (PR)-driven bioassays to track changes in serum progestogenic activity following administration of traditional Chinese medicinal herb, Ligusticum chuanxiong, with potent progestogenic activity. Sensitive and specific (>300-fold) increases in progestogenic activity were observed when HeLa cells transfected with PR and a PR-driven promoter were exposed to the progestogenic drug, medroxy-progesterone acetate (MPA), suggesting the utility of the bioassay to measure progestogenic effects for PK/PD studies. Progestogens were administered to male Sprague-Dawley rats and serum extracted for measurement of progestogenic activity. Effect-time studies indicate that injection of MPA and L. chuanxiong extract raised area-under-curve of progestogenic activity in sera by 8.2-fold (p<0.001) and 4.5-fold (p<0.01) respectively, compared to sera from rats administered vehicle only. Administration of MPA and L. chuanxiong extract by the oral route resulted in a 5.4 (p<0.001) and 2.3-fold (p=0.07) increase respectively. Our data suggest that PR-responsive reporter gene bioassays can measure bioavailability of compounds, known and unknown, of complex botanicals for hormone replacement therapy. L. chuanxiong extracts exert progestogenic activity in vivo, and may have utility for progesterone-replacement therapy.

[Research on effect of total saponins of yinfenglun on uterine].

OBJECTIVE: To research the effect of total saponins of Yinfenglun on uterine. METHOD: Models of uterine inflammation were established to observe the effect of total saponins of Yinfenglun. Uterine contractive effects were studied on rats in vitro and on rabbit in vivo. Weight of uterus and levels of estrogen and progestogen were determined. RESULT: Total Saponins of Yinfenglun had the ameliorated tendency to metritis of model rats, and increased the contractive range and motorricity of uterine of rats in vitro and of rabbit in vivo. The effect to uterus in vivo maintained longer. Total saponins of Yinfenglun could increase the weight of uterus and have an increased tendency on the content of estrogen, but not the level of progestogen. CONCLUSION: There are obvious effects on uterine of total saponins of Yinfenglun, which are related to its clinical use.

[Clinical observation on treatment of Stein-Leventhal syndrome caused sterility by combined use of clomiphene and Chinese nourishing shen and activating blood circulation drugs].

OBJECTIVE: To observe the efficacy of combination therapy of clomiphene and Chinese drugs for nourishing Shen and activating blood circulation (NSABC) in treating Stein-Leventhal syndrome caused sterility. METHODS: Sixty-two patients with anovulation caused sterility were randomly divided into the treated group (n = 32) and the control group (n = 30). The treated grop was treated with the combination therapy and the control group treated by the same dosage of clomiphene alone. RESULTS: After treatment, when comparing with that before treatment, the endocrine hormones in the treated group improved significantly, showing a markedly decrease of androgen and luteotropic hormone, and increase of estrogen (P < 0.001). The periodic ovulation rate in the treated group reached 87%, the total pregnancy rate being 65.6%, with no occurrence of ovarian hyperstimulation syndrome (OHSS) and luteinized unruptured follicle syndrome (LUFS), while in the control group, the periodic ovulation rate was 66%, the total pregnancy rate 36.6%, with LUFS occurred in 4 patients. Comparison of the therapeutic effects between the two groups showed significant difference (P < 0.05). CONCLUSION: The combination therapy of clomiphene and NSABC has a better therapeutic effect in treating Stein-Leventhal syndrome caused sterility than that of using clomiphene alone.

[The effects of herb lithospermum extract on MCF-7 cell and estrogen and progestogen levels in mice].

OBJECTIVE: To study the effects of lithospermum extract on MCF-7 cell and estrogen and progestogen levels in mice. METHOD: Cell growth curve and Western Blotting were used to do animal experiment. RESULT: Lithospermum extract could inhibit the growth of MCF-7 cell. It could also inhibit the expression of ER and increase the expression of PR with large dose. After the mice were bred with Lithospermum, their serum estrogen and progestogen levels reduced, their uterus weight index decresed and uterus ER and PR levels increased. It could also improve the hyperplasia of uterus caused by tamoxifen. CONCLUSION: Lithospermum extract can inhibit the growth of MCF-7 cell and inhibit the level of estrogen and progestogen in mice.

Six-month oral dehydroepiandrosterone supplementation in early and late postmenopause.

The adrenal production of the delta 5-androgens, dehydroepiandrosterone (DHEA) and its sulfate ester dehydroepiandrosterone sulfate (DHEAS), declines linearly with aging. The evidence that DHEA or DHEAS administration may alleviate some of the problems related to aging has opened new perspectives for clinical research. The present study aims to investigate the effects of a 6-month DHEA supplementation in early and late postmenopausal women, with normal or overweight body mass index (BMI), on the level of circulating steroids, sex hormone binding globulin (SHBG), beta-endorphin and gonadotropins, and on the adrenal gland response to dexamethasone suppression and adrenocorticotropic hormone (ACTH) stimulation. Early postmenopausal women (50-55 years) both normal weight (BMI 20-24, n = 9) and overweight (BMI 26-30, n = 9) and late postmenopausal women (60-65 years) both of normal weight and overweight, were treated with oral DHEA (50 mg/day). Circulating DHEA, DHEAS, 17-OH pregnenolone, progesterone, 17-OH progesterone, allopregnenolone, androstenedione, testosterone, dihydrotestosterone, estrone, estradiol, SHBG, cortisol, luteinizing hormone, follicle stimulating hormone and beta-endorphin levels were evaluated monthly and a Kupperman score was performed. The product/precursor ratios of adrenal steroid levels were used to assess the relative activities of the adrenal cortex enzymes. Before and after 3 and 6 months of therapy, each women underwent an ACTH stimulating test (10 micrograms i.v. in bolus) after dexamethasone administration (0.5 mg p.o.) to evaluate the response of cortisol, DHEA, DHEAS, androstenedione, 17-OH pregnenolone, allopregnanolone, progesterone and 17-OH progesterone. The between-group differences observed before treatment disappeared during DHEA administration. Levels of 17-OH pregnenolone remained constant during the 6 months. Levels of DHEA, DHEAS, androstenedione, testosterone and dihydrotestosterone increased progressively from the first month of treatment. Levels of estradiol and estrone significantly increased after the first/second month of treatment. Levels of SHBG significantly decreased from the second month of treatment only in overweight late postmenopausal women, while the other groups showed constant levels. Progesterone levels remained constant in all groups, while 17-OH progesterone levels showed a slight but significant increase in all groups. Allopregnanolone and plasma beta-endorphin levels increased progressively and significantly in the four groups, reaching values three times higher than baseline. Levels of cortisol and gonadotropins progressively decreased in all groups. The product/precursor ratios of adrenal steroid levels at the sixth month were used to assess the relative activities of the adrenal cortex enzymes and were compared to those found before therapy. The 17,20-desmolase, sulfatase and/or sulfotransferase, 17,20-lyase and 5 alpha-reductase activities significantly increased, while the 3 beta-hydroxysteroid-oxidoreductase activity did not vary. On the contrary, the 11-hydroxylase and/or 21-hydroxylase activities showed a significant decrease after 6 months of treatment. In basal conditions, dexamethasone significantly suppressed all the adrenal steroids and this suppression was greater after 3 and 6 months of treatment for DHEA, DHEAS and allopregnanolone, while it remained unchanged for other steroids. Before treatment, ACTH stimulus induced a significant response in all parameters; after the treatment, it prompted a greater response in delta 5- and delta 4-androgens, progesterone and 17-OH progesterone, while cortisol responded less in both younger and older normal-weight women. The endometrial thickness did not show significant modifications in any of the groups of postmenopausal women during the 6 months of treatment. Treatment with DHEA was associated with a progressive improvement of the Kupperman score in all groups, with major effects on the vasomotor symptoms in

Effect of the aromatase inhibitor, 4 hydroxyandrostenedione, on the endotoxin-induced changes in steroid hormones in male rats.

The increase in circulating estrogen concentrations that follows injection of Escherichia coli endotoxin (Endo) may be due to increased aromatase activity. We have therefore analysed the effect of the aromatase inhibitor, 4 hydroxyandrostenedione (4OHA) on the steroid hormone response of male rats, particularly the dramatic increase in estrogens and decrease in androgens, induced by Endo. The concentrations of corticosterone (B), progesterone (P4), 17 alpha hydroxyprogesterone (17 alpha OHP4), androstenedione (delta 4), testosterone (T), estrone (E1) and estradiol (E2) were determined 2 hours after injection of increasing doses of 4OHA with and without Endo. The increase in serum estrogen concentrations and drop in serum androgen levels in response to Endo were blocked by a single dose of 4OHA. The effect of 4OHA appeared to be dose dependent. Low doses (30 mg/kg and 50 mg/kg) induced significant changes in the estrogen and androgen responses, but the high dose (100 mg/kg) blocked all changes in sex steroids induced by Endo. 4OHA did not alter the Endo-induced changes in other steroids.

Antifertility profile of the aqueous extract of Moringa oleifera roots.

An aqueous extract of Moringa oleifera roots was investigated for its estrogenic, anti-estrogenic, progestational and antiprogestational activities. Oral administration of extract progressively increased the uterine wet weight of bilaterally ovariectomized rats. This estrogenic activity was supported by stimulation of uterine histo-architecture. When the extract was given conjointly with estradiol dipropionate (EDP), there was a successive reduction in the uterine wet weight when compared to the gain with EDP alone and uterine histological structures were also inhibited. In the deciduoma test, the highest dose of 600 mg/kg interfered with the formation of deciduoma in 50% of the rats, showing some antiprogestational activity. Doses up to 600 mg/kg of the extract orally failed to induce a decidual response in the traumatized uterus of ovariectomized rats. The antifertility effect of the extract appears to be due to multiple attributes.

[Effects of anterior hypothalamic deafferentation on plasma LH response to LH-RH in the baboon (author's transl)].

We reported previously that a biphasic LH release (within 30 min and 90 approximately 150 min after LH-RH injection) was observed in normal menstruating baboons (non-human primates). Plasma immunoassayable LH-RH reached a maximum within 4 min after injection and was undetectable within 60 min. Plasma estrogen and progestin were elevated within 45 min. No plasma LH release with increment of plasma estrogen and progestin was observed in saline injected baboons. This study was performed to investigate the mechanism of biphasic LH release by LH-RH injection. Synthetic LH-RH (100 microgram) was injected sc into four female retrochiasmatic deafferented baboons. Blood samples collected 30 and 2 min before injection and 5, 10, 15, 30, 45, 60, 90, 120, 150 and 180 min after injection were assayed for LH, estrogen and progestin. In four baboons plasma LH peak was observed within 30 min after injection and plasma estrogen and progestin were elevated within 45 min. However, plasma LH peak within 90 approximately 150 min was not observed. These results infer that exogenous LH-RH exerts an effect on the pituitary to release LH within 30 min after injection. Increased estrogen and progestin exert the effect on the higher brain area (extrahypothalamic area) and the anterior hypothalamus to facilitate release of endogenous LH-RH, which subsequently release LH within 90 approximately 150 min in cooperation with exogenous LH-RH which may be considered to participate LH synthesis in the pituitary. Thus, it seems likely that the extrahypothalamic area (for example; limbic system) and the anterior hypothalamus have an important role in regulating LH-RH secretion and subsequent LH release in the baboon.