The Effect Of Guiling Xiaoyi Decoction Drug Serum On The Secreting Of BCL-2FAS On Eupopic Glandular-cell In Endometriosis

Endometriosis is a common and frequently-occurring gynecological disease which defined as endometriotic tissues growing outside the normal uterine cavity.The benign disease can distant metastasis and growth like malignant disease. The incidence has showed rising trend in recent years.Endometriosis is one of the most common gynecologic disease. Endometriosis often causes dysmenorrhea, dyspareunia and infertility.At present the principal treatment is hormoual therapy and surgical.But false pregnancy and false postmenopausal phenomenon with long-term hormone treatment and complication with surgical that cause many advese effects for patients physical and mental health.According to zheng differentiation-treatment,trditional Chinese medicine can release clinical symptoms and regulate menstruation. Guizhi fuling pill originates from Jinkui yaolue. Zhongjing Zhang, the author of the book,designed this prescription to mildly remove mass in the abdomen,disperse blood stasis so as to prevent abortion which was due to the abdominal mass from endangering the fetus.If the prescription was utilized mainly in order to remove blood stasis and disperse the mass in the abdomen,there may be some shortage of the strength of the prescription. So,first more drugs having the property of enhancing blood circulation and resolving blood stasis should be added to the prescription.Second, the prescription should be modified according to syndrome differentiation. Therefore,Guiling xiaoyi Decoction was designed which combines the Guizhi fuling pill with Xianlingpi to strengthen warming kidney, and Xuejie to disperse blood stasis.Objective:To investigate the theoretical foundation of Guiling xiaoyi decoction on theating Endometriosis,as well as the expression of Bcl-2Fas induce by durg serum of Guiling xiaoyi decoction on glandular cultured in-vitro. Methods:The eutopic endometial glandular cells were isolated and cultured in vitro.The proliferation were obseved by inverted microscope.Then they were ideneified by vimentin immunocytochemistry staining.The serums of different dosage drugs of Guiling xiaoyi decoction inferfered the glandular cells.The growth and inhibition of eutopic glandular cells were measured by MTT assay.Collecting supernatant liquid of cell culture at the time of24h,48h,72h,respectively detected the concentration of Bcl-2,Fas by the method of ELISA.Result:Human endometrial glandular epithelial cells could be successfully isolated and cultured by combination of enzyme digestion, sieve separation, centrifugal separation and differential adhesion method, with88.89%success rate. The cells could be subcultured for1-2generations, and survived for4-6weeks. The glandular epithelial cells from both endometriosis and non-endometriosis can be isolated and cultured successfully(.P>0.05). Furthermore, the derived time of endometrial glandular epithelial cells doesn’t have an influence both in proliferative advanced stage and secretory phase on the survival rate and proliferative capacity (P>0.05). An effective research platform can be provided for the study on endometriosis pathogenesy and pharmacodynamics.Through fixed detection, we confirmed that the serums of Guiling xiaoyi Decoction could reduce Bcl-2secretion, and increase Fas secretion of endometrial glandular epithelial cells(P<0.01), and the effect of inhibition is more and more obvious with the concentration increasing and the time extending.Conclusion:1. Bcl-2and Fas are all expressed in both normal endometrium and eutopic endometrium of endometriosis. Expression of Bcl-2in proliferative phase is higher than that in secretory phase of normal endometrium glandular epithelial cells. But expression of Fas in secretory phase is higher than that in proliferative phase. Expression of Bcl-2in eutopic endometrium of EMs is higher than that in normal endometrium, and this high expression have no relationship with the different phases of menstrual cycle. Expression of Fas in eutopic endometrium of EMs is lowerr than that in normal endometrium, and this low expression also have no relationship with the different phases of menstrual cycle.2.Bcl-2secretion of cell culture supernatant is significantly reduced through Guiling xiaoyi Decoction interfering in endometrial glandular epithelial cells of patients with EMs(P<0.01), and the extent of reduction is more and more obvious with the concentration increasing and the time extending.3.Fas secretion of cell culture supernatant is significantly increased through Guiling xiaoyi Decoction interfering in endometrial glandular epithelial cells of patients with EMs(P<0.01), and the extent of increase is more and more obvious with the concentration increasing and the time extending.