The Study Of The Effect Of Bushen Zhuyun Decoction On End- Ometrial Cells ER、MMP-9、TIMP-1 And Ovarian Secretion Hormones

Objective:Observed BuShenZhuYun Decoction-containing on rat luteal phase endometrial cells of estrogen receptor (ER), matrix metalloproteinase-9 (MMP-9), matrix metalloproteinase inhibitor-1 (TIMP-1) intervened by mifepristone and on the regulation of rat granulosa cells in luteal estradiol (E2), progesterone (P) levels of influence. Discussion BuShenZhuYun Treatment luteal infertility effective mechanism.Method:(1)Preparation of serum-containing drugs:used an ordinary male rabbits, to BuShenZhuYun herbs decoction 40g/kg orally (equivalent to 10 times the adult clinical dose),2 times/d, continuous 3d. Fourth day of the last administration 1h (before gavage fasted 12h), a local anesthetic carotid blood, at room temperature for 3h,3000rpm centrifuge serum 15min, serum was collected,-20℃ preservation,56℃ water bath to inactivate complement 30min, 0.22um filter membrane sterilized before use serum-free DMEM diluted to5%,20% concentration containing serum.(2)Endometrial cells, ovarian luteal cells isolated and cultured: Select five weighing 50-60g underage female rats, subcutaneous PMSG 50U/only,65h ovarian follicle after subcutaneous injection of HCG 35U/rats injected seventh days (HCG injection day of the first day), pull the neck marrow were killed off, the removal of the ovaries, uterus, isolated and cultured endometrial cells and ovarian luteal cells. After phenotypic identification of endometrial cells, ovarian luteal cell cultures containing 100U/ml penicillin, 100U/ml streptomycin and 10% fetal bovine serum, DMEM culture medium, placed in 37℃,5% CO2 incubator training, daily observe the growth of adherent 3-5d passaged once. Subculturing 0.25% trypsin, DMEM culture medium.(3)Containing serum intervention:intervention plus medication 48h, cell protein extracts of each group, RNA and cell supernatants were observed BuShenZhuYun contained serum endometrial cells ERα, ERβ, MMP-9, TIMP-1 and ovarian granulosa cells secrete estrogen and progesterone levels. Western blot was used to detect endometrial cells ER α,ERβ protein expression, RT-PCR was used to detect endometrial cells ERα,ERβ, MMP-9, the expression of TIMP-1 mRNA of radioimmunoassay luteinizing granulosa cell supernatants were female, progesterone hormone levels.Result:(1)BuShenZhuYun Decoction-containing improved the expression of ERα, ERβ protein on endometrial cells; while significantly improving the low expression of ERα,ERβ protein mifepristone-induced;(2)BuShenZhuYun Decoction-containing improved endometrial cells ER, MMP-9,TIMP-1 transcriptional gene expression;while significantly improving the mifepristone-induced ER,MMP-9,TIMP-1 in low expression;(3)BuShenZhuYun Decoction- containing significantly improved the luteal phase of the ovarian granulosa cells secrete estrogen and progesterone levels.Conclusion:(1)BuShenZhuYun Decoction increased the endometrial cells ER, MMP-9, TIMP-1 expression, improved endometrial receptivity conducive to embryo implantation pregnancy, laying the foundation for embryo implantation;(2)BuShenZhuYun Decoction increased the secretion of E2, P levels on ovarian luteal cells, improved mifepristone-induced E2, P low expression levels, improved mifepristone-induced corpus luteum function decline, improved luteal function in favor of embryonic implantation and pregnancy to maintain and improved the probability of clinical pregnancy.