Kidney-Replenishing Herb Induces SOCS-3 Expression via ERK/MAPK Pathway and Improves Growth of the First-Trimester Human Trophoblast Cells.

Kidney-replenishing herb is a traditional medicine formula in China which has been widely used for clinical treatment of recurrent miscarriage. Our previous study showed that Kidney-replenishing herb could promote proliferation and inhibit apoptosis of the human first-trimester trophoblasts. In the present study, we further explored the potential mechanism and signal pathway of Kidney-replenishing herb on human trophoblast cells. Our research showed that Kidney-replenishing herb stimulated proliferation and reduced apoptosis of human trophoblast cells in vitro, and this appeared to be positive correlation with SOCS-3 transcription, suggesting that Kidney-replenishing herb regulated biological functions of human trophoblast cells by inducing SCOS-3 expression. Furthermore, the Kidney-replenishing herb treatment stimulated the phosphorylation of ERK1/2, and blocking the signaling pathway by mitogen-activated protein MAPK (MEK) inhibitor, U0126, inhibited Kidney-replenishing herb-induced SOCS-3 transcription, depressed proliferation, and promoted apoptosis of human trophoblasts. Kidney-replenishing herbs still induced ERK1/2 phosphorylation after SOCS-3 siRNA silence. Overexpression of SOCS-3 stimulated the proliferation of trophoblast. These findings suggest that SOCS-3 expression is induced by Kidney-replenishing herbs via activation of MAPK pathways, and this may possibly be involved in promoting human trophoblast cells growth which is contributed to embryo development.

Efficacy of Chinese patent medicine Tian Gui Capsule in patients with polycystic ovary syndrome: a randomized controlled trial.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a complex hormonal disorder and one of the most common reproductive endocrinology abnormalities in women. Recently, many studies have been conducted assessing Chinese herbal medicine as an alternative treatment for women with PCOS, it is, therefore, worthwhile to analyze and observe the curative effects of traditional Chinese medicine treatment in PCOS. OBJECTIVE: To evaluate the efficacy of the Chinese patent medicine Tian Gui Capsule, in women with PCOS and compare its effects with metformin and ethinyl estradiol plus cyproterone acetate (Diane-35). DESIGN, SETTING, PARTICIPANTS AND INTERVENTION: A total of 47 PCOS outpatients from the Obstetrics and Gynecology Hospital of Fudan University were randomly divided into 3 groups. Patients in group A (n=19) were given Tian Gui Capsule, patients in group B (n=17) were given metformin, and patients in group C (n=11) were given Diane-35. The 3 groups of patients were treated for 3 months. MAIN OUTCOME MEASURES: Serum testosterone (T), sex hormone binding globulin (SHBG) and dehydroepiandrosterone sulfate (DHEA-S) levels, free androgen index (FAI), fasting blood glucose (FPG), fasting insulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR), insulin sensitive index (ISI) and left and right ovary volumes of the 3 groups were evaluated before and after treatment . RESULTS: After 3 months of treatment, when compared with before treatment data, group A patients showed decreased serum T and SHBG levels, FAI, FINS, and left and right ovary volumes (P<0.05), and increased serum DHEA-S (P<0.05), while the FPG level showed no significant change. Although the level of serum T and FINS among the 3 groups after the treatment were similar, group A demonstrated better results than group B in reducing the FAI and increasing the serum SHBG, but less significant results than group C besides, group B was the only group showed improved insulin sensitivity. Although the level of FPG of the 3 groups after treatment were similar, group C had the most increased FPG. CONCLUSION: The effects of Tian Gui Capsule on hyperandrogenism are not as significant as Diane-35, but more effective than metformin. The effects of Tian Gui Capsule on hyperinsulinemia are not as significant as metformin but better than Diane-35. Tian Gui Capsule treats PCOS by regulating ovarian functions and reducing blood insulin level without inhibiting the function of the hypothalamic-pituitary-ovarian axis. Further studies with larger sample size are needed to confirm the above results.

[Prelimary exploration on Chinese medicine syndrome type distribution in patients with polycystic ovary syndrome].

OBJECTIVE: To explore the Chinese medicine syndrome type distribution in patients with polycystic ovary syndrome (PCOS) and its relationship with sexual hormones. METHODS: Chinese medicine syndrome types of 212 PCOS patients were differentiated and sorted by adopting fuzzy mean C clustering method, and their relationship with the indices of sexual hormones detected on the 3rd to 5th day of menstrual cycle was analyzed, with the values got from 20 healthy women for controls. RESULTS: Intermingling syndromes were commonly seen in PCOS patients. Shen-deficiency syndrome (presented in 64 patients) and Gan-qi stagnancy syndrome (61 patients) were the dominance, accounting for 30.2% and 28.8% respectively, significantly higher than that of other syndromes (P < 0.05), which were Pi-deficiency syndrome (41 patients, 19.3%), phlegm-dampness syndrome (33 patients, 15.6%) and blood stasis syndrome (13 patients, 6.1%). Levels of estradiol (E2), testosterone (T), luteinzing hormone (LH), dehydroiso-androsterone (DHEA-S) and prolactin (PRL) were higher, while the level of sexual hormone binding protein (SHBG) was lower in PCOS patients than those in control, follicular stimulating hormone (FSH) level in patients of Shen-deficiency syndrome and phlegm-dampness syndrome was high than that in control (P < 0.05 and P < 0.01). However, no significant differences were found in comparing the various sexual endocrinal indices between patients with different syndrome types (P > 0.05). Besides, the level of PRL was positively correlated with LH and E2 levels in patients. CONCLUSION: Chinese medicine syndromes presented in patients with PCOS are mostly intermingling, Shen-deficiency and Gan-stagnancy are the basic syndromes, and there is some correlation between syndrome type and sexual hormone levels.

HPV E6 down-regulation and apoptosis induction of human cervical cancer cells by a novel lipid-soluble extract (PE) from Pinellia pedatisecta Schott in vitro.

AIM OF THE STUDY: To evaluate the cytotoxicity and apoptosis induction effects of a novel lipid-soluble extract (PE) from Pinellia pedatisecta Schott on CaSki, HeLa and HBL-100 cells. Particularly, the effect of PE on HPV E6 gene expression was tested, and the mechanism of its apoptosis induction effect was also studied. MATERIALS AND METHODS: Cell viability was measured by the MTT assay. DAPI staining and flow cytometric analysis (FCM) were used to identify apoptotic cells in PE-treated CaSki, HeLa, and HBL-100 cells. Expression of the HPV E6 gene in CaSki and HeLa cells was detected by real-time RT-PCR and western blot analysis. Apoptosis-associated genes were examined by RT-PCR and western blot analysis in CaSki cells. RESULTS: PE inhibited the growth of CaSki and HeLa cells in a time- and dose-dependent manner, but it had no obvious inhibiting effect on HBL-100 cells except at a relatively high dose (500 µg/mL). PE could induce apoptosis in CaSki and HeLa cells in a time-dependent manner but not in HBL-100 cells. HPV E6 mRNA and protein were decreased significantly by PE. Caspase-8, caspase-3, Bax, P53 and P21 mRNAs as well as proteins were increased while Bcl-2 mRNA and protein were decreased significantly by 24 h of PE treatment. CONCLUSIONS: PE can function as a tumor suppressor by inducing apoptosis in human cervical cancer cells but it has little side effect on normal cells. It probably acts via mitochondria-dependent and death receptor-dependent apoptotic pathways. HPV E6 may be the key target of its action.

[Effect of pinellia extract on HeLa cell line in vitro and associated mechanism].

OBJECTIVE: To investigate the effect of Pinellia extract (PE) on HeLa cell line and to study its associated mechanisms, in order to provide theoretical foundations for its applying in clinical prevention and treatment of cervical disease. METHODS: HeLa cell line was incubated in media containing different concentrations of PE. Growth of cells was observed and photo-generated with inverted phase microscope; cell activity was detected by MTT assay; cell apoptosis detected by flow cytometry, protein expression of proliferating cell nuclear antigen proliferating cell nuclear antigen (PCNA) was detected by immuno-cytochemistry, and protein expression of Bcl-2 was determined by immunofluorescence. RESULTS: PE showed obvious inhibition on the proliferation of HeLa cells, cell apoptosis appeared after PE treatment in a time and dose dependent manner; PCNA and Bcl-2 protein expressions reduced significantly after being effected by PE for 24 h. CONCLUSIONS: PE can obviously inhibit the growth and induce the apoptosis of HeLa cells, and its mechanism is possibly realized through down-regulating the expressions of PCNA and Bcl-2. The study set a primary experimental base for further studying the action mechanism of PE and developing new anti-tumor agents.

[Regulation of vitality and IL-10 receptor expression of human decidual stromal cells in early pregnancy by IL-10, IFN-gamma].

The study is to investigate the effect of IL-10, IFN-gamma on viability and IL-10 receptor expression of human decidual stromal cells (DSC) in early pregnancy. The vitality of DSC was detected by MTT. IL-10R1 and IL-10R gene expression of DSC were detected after treated by different concentrations of IL-10, IFN-gamma at 15min, 30min, 45min, 60min. The vitality of DSC was significantly enhanced by the higher dose of IL-10 (100-10ng/ml) (P<0.05), and there was no significant stimulative role induced by the lower dose of it(0.10-1ng/ml)(P>0.05). The viability of DSC was inhibited by the highest dose of IFN-gamma (1000ng/ml) (P<0.01), on the contrary, its viability was stimulated by the lower dose of IFN-gamma (10ng/ml) (P<0.05). IL-10R1 was highly expressed by IL-10 of 10ng/ml within 15min, and significantly reduced at 30min, then disappeared within 45min; while IL-10R1 expression was not induced by IL-10(1ng/ml) of lower concentration throughout 60min; IL-10R1 was briefly and lowly expressed by IFN-gamma of 100ng/ml, which effect on DSC at 45min; The expression of IL-10R1 was moderate level induced by IFN-gamma of 10ng/ml within 30min,and reduced at 45min,then disappeared at 60min. There was no significant difference of IL-10R2 expression (P>0.05) between treatment and not with the above-mentioned cytokines. IL-10, IFN-gamma may play an important role of immune regulation in early pregriancy by influencing IL-10R1 expression and vitality of DSC.

[Effects of beta-sitosterol on microtubular systems in cervical cancer cells].

OBJECTIVE: To investigate beta-sitosterol's inhibitory effects on SiHa cells' growth, and the effects on microtubular system in SiHa cell. METHODS: Proliferation inhibition of SiHa cell line was evaluated by MTT assay. Cell cycle of SiHa cells treated with beta-sitosterol was analyzed by flow cytometry. The expression and distribution of microtubule and microtubule associated protein 2 in SiHa cells were investigated by confocal microscopy. Immunoblotting analysis was used to determine tubulin alpha, microtubule associated protein 2, and the proportion of polymerization of tubulin. RESULTS: beta-sitosterol could obviously inhibit the proliferation of SiHa cells, and induce the accumulation of cells in S phase (rather than the G2/M phase) and mitotic arrest in the cell cycle. Confocal microscopy showed an abnormal microtubular network in SiHa cell treated with beta-sitosterol for 5 days, and the expression of microtubule associated protein 2 was marked down-regulated. Further analysis by immunoblotting confirmed the down-regulation of beta-sitosterol on the expression for both microtubule associated protein 2 and tubulin alpha. Moreover, beta-sitosterol reduced the proportion of polymerization of microtubule in a time-dependent manner. CONCLUSION: beta-sitosterol could down-regulate the expression of tubulin alpha and microtubule associated protein 2 in SiHa cells, and inhibit the microtubular polymerization. Our results suggested an anti-microtubule characteristic of beta-sitosterol which might contribute to the proliferation inhibition of SiHa cells.

[Effect of bushen yiqi recipe on bioactivity behavior of human cytotrophoblast of early pregnancy].

OBJECTIVE: To investigate the effect of Chinese drugs for tonifying Shen on the bioactivity, cell proliferation, invasion and differentiation of human cytotrophoblast of early pregnancy. METHODS: The human early pregnant cytotrophoblast was cultured in vitro, and treated with saline (as blank control) and drug containing serum in different concentrations (5%, 10% and 20%) respectively. The changes of morphology, proliferation and invasive capacity of cells were examined by scanning electron microscopy, MTT method, flow cytometry, Transwell invasive assay at 24 hrs, 48 hrs and 72 hrs after treatment. RESULTS: After cells being cultured with drug containing serum, the cytotrophoblast became abundant in microvilli, with more and prolonged pseudopodia. The cell absorbency in 490 nm wave length increased significantly (P < 0.01), cells of sub-G1 and G2/M phase obviously decreased and that of S phase increased (P < 0.01), and the cells penetrated through PET membrane in each visual field significantly increased (P < 0.01). CONCLUSION: Chinese herbs for tonifying Shen could promote the proliferation and invasive capacity of cytotrophoblast and might influence its differentiation.

[Effect of Chinese herbal treatment on the efficiency of blocking CD antigen and on prolactin and progesterone in women with recurrent spontaneous abortion].

OBJECTIVE: To explore the effect of Chinese herbal medicine Baotai Granule (BTG, a self-made preparation) on CD antigen blocking efficiency, prolactin (PRL) and progesterone (P) in patients with recurrent spontaneous abortion (RSA). METHODS: Thirty-four women suffered from RSA were treated with BTG, twice every day, 1 package (10 g) in each time by orally intake. Changes of the efficiency of serum blocking antibody in them to the CD antigen in their husband's peripheral T-lymphocytes before and after treatment were observed. And the changes of blood levels of PRL and P were also monitored. RESULTS: Fetus had successfully protected in 30 women (88.2%), in them, the efficiency of blocking to CD3, CD4 and CD8 after treatment were all higher than that before treatment, and levels of PRL and P in peripheral blood increased along with the increase of gestational age, while no obvious change was found in those who failed to complete pregnancy. CONCLUSION: Chinese herbal medicine could protect the fetus by regulating the response between endocrine and immunity network during pregnancy.