Investigate The Role Of "Pingchongjiangnifang"of The In Vitro Culture Of Human Endometrial Stromal Cells

Objective:Establishment of human endometrial stromal cell model to explore the Chinese medicine “ Pingchongjiangnifang ”of endometriosis of the mechanism of action from cytological level.Method:Centrifugation,culture and identification of endometrial stromal cells by mesh filtration combined with collagenase digestion and separation,to establish human endometriosis disease stromal cell model of endometriosis and non-endometriosis group.Chinese Herbs “ Pingchongjiangnifang ”concentrate(Guizhi 15 g,Xuejie Powder 3g,Wulingzhi vinegar 10 g,Puhuang cloth 6g,Chenxiang after the next 6g,Shuizhi Talc fried3 g,Baishao 10 g,Maidong10g,Gancao6g),mifepristone and saline lml/100 g were given to gavage for 7 days,2 times daily.Then the traditional chinese drug-containing,western drug-containing serum,and blank-containing serum were added to cultured stromal cells,Measured using the MTT method assay the survival of mesenchymal cells,calculate OD values between mesenchymal cells to observe the increase,calculate the through cell number to observe migration and invasion with Transwell experiment;observe the apoptosis with flow cytometry apoptosis rate.Results:1.Compare the absorbance values of endometrial stromal cells after different affect of containing serum in 24,48,72 h.Compared with the control group,at the same time,the traditional chinese drug-containing of endometrial stromal cell proliferation in the most obvious difference was statistically significant(P<0.05);different times,proliferation inhibition is best in 48 h,the difference was statistically significant(P <0.05).2.Compare the endometriosis endometrial stromal cell migration with non-endometriosis patients,the number of endometrial stromal cells migrating within endometriosis and non-endometriosis after the treatment with traditional chinese drug-containing were less than blank serum,and endometriosis group is better than non-endometriosis group,the difference was significant(P <0.05).20% of the number of cells migrating is least.3.Comparison of endometrial stromal cell invasion of endometriosis and non-endometriosis patients,the number of endometrial stromal cells invasive within endometriosis and non-endometriosis after the treatment with traditional chinese drug-containing were less than blank serum,and endometriosis group is better than non-endometriosis group,the difference was significant(P <0.05).20% of the number of cells invasive is least.4.Flow cytometry was performed on cells treated with different treatment to analyze the apoptosis detection showed that serum treatment can make EMs and non-EMs endometrial stromal cell apoptosis,apoptosis rates of the blank serum below western medicine serum,western medicine serum is lower than traditional chinese drug-containing serum,and the endometrial stromal cells of EMs is higher than non-EMs endometrial.the difference was significant(P <0.05).Conclusion:1 Pingchongjiangnifang can inhibit the in vitro culture endometrial stromal cell proliferation,migration,invasion behavior,and promote cell apoptosis;2 Pingchongjiangnifang effects on cell proliferation in dose time dependence;3 Pingchongjiangnifang effects on endometrial stromal cell proliferation,migration and invasion of endometriosis is stronger than normal endometrial stromal cells,and cell apoptosis is weaker than normal endometrial stromal cells;4 Compared with mifepristone,Pingchongjiangnifang on cell proliferation,migration and invasion has greater inhibition,and apoptosis has a greater role in promoting,Tip: Pingchongjiangnifang may be more effective than mifepristone in EMs drug treatment on aspects of cell affected,provide a new theoretical basis for Pingchongjiangnifang treat Ems.