| Background and purposeSpontaneous abortion is a common complication of pregnancy.According to the different stages of spontaneous abortion,it can be divided into threatened abortion,abortion,incomplete abortion and complete abortion.Treatment of patients with spontaneous abortion in a timely manner can further affect the outcome of pregnancy.Therefore,the study of the prevention and treatment in spontaneous abortion has a certain social significance.The book Records of Tradition Chinese and Western Medicine in Combination was written by Zhang Xichun who was a physician in Qing Dynasty and the formulae Shoutai Pill was recorded in this book.In recent years,the formulae is widely used in the prevention and treatment in spontaneous abortion.Early experimental study found that the serum containing Shoutai Pill in a dose-dependent manner to enhance the activity of proliferation,the capacity of invasion and migration,reduce apoptosis of trophoblastic cells in patients with spontaneous abortion.Based on this,this research explored the effects of the serum containing Shoutai Pill on the apoptosis and the expression of Bcl-2,Bax,Fas,FasL and Survivin in trophoblastic cells in patients with spontaneous abortion,and to further explore the mechanism of Shoutai Pill in prevention and treatment in spontaneous abortion and its target.Methods1.A total of 36 SPF grade SD rats were randomly divided into three groups:Shoutai Pill water decoction(6.25g/kg),dydrogesterone normal saline solution(2.08 mg/kg),equal volume of normal saline by oral administration.Make preparation of the blank serum,the serum containing Shoutai Pill and dydrogesterone.2.1n vitro isolate and culture trophoblastic cells(TB)of the patients witih spontaneous abortion and normal pregnant women experienced artificial abortion surgery.The expression of Cytokeratin-7(CK-7)and Vimentin were detected by immunocytochemistry experiments.After confirming as trophoblastic cells,the follow-up experiments continued.3.Intervent with trophoblastic cells of the patients with spontaneous abortion in different concentrations(5%,10%,20%)of the serum containing Shoutai Pill.Blank serum and serum containing dydrogesterone were taken as the blank control and the positive control,respectively.4.Flow cytometry(FCM)was used to detect the early apoptosis rate and the total apoptosis rate of trophoblastic cells in each group.5.The expression of Bcl-2,Bax,Fas,FasL and Survivin were detected by Western blot,and the correlation between the expression of Bcl-2,Bax,Fas,FasL and Survivin was analyzed.6.Statistical methods:SPSS software version 20.0 was used for analysis.All the measurement data were expressed as means±SD.According to the different experimental design,different statistical method was used.P<0.05 for the difference was statistically significant.Results1.Compared with the normal control group,the early apoptosis rate and the total apoptosis rate of trophoblastic cells of spontaneous abortion group was significantly higher(P<0.05).2.Compared with the normal control group,the level of Bcl-2 and Survivin protein in the trophoblastic cells of spontaneous abortion group was significantly lower,and the level of Fas,FasL and Bax was significantly higher(P<0.01).3.Compared with the blank control group,the early apoptosis rate and the total apoptosis rate of trophoblastic cells in the patients with spontaneous abortion was significantly lower in the group which was intervened with the serum containing Shoutai Pill and dydrogesterone(P<0.01).4.Compared with the blank control group,the levels of Bcl-2 and Survivin protein were up-regulated and the levels of Fas,FasL and Bax were down-regulated after the intervention of the serum containing Shoutai Pill and dydrogesterone(P<0.01).5.The serum containing Shoutai Pill had a dose-dependently effect in the early apoptosis rate,the total apoptosis rate and the expression of five kinds of apoptosis protein of the patients with spontaneous abortion(P<0.05).6.There was a significant positive correlation between Bcl-2 and Survivin.There was a significant negative correlation between Bax,Fas,FasL and Survivin(P<0.01).ConclusionThe serum containing Shoutai Pill can dose-dependently reduce the apoptosis of trophoblastic cells in the patients with spontaneous abortion,up-regulate the expression of Bcl-2 and Survivin,down-regulate the expression of Fas,FasL and Bax,inhibit the apoptosis of trophoblastic cells through the intrinsic and extrinsic pathways.It maybe one of the possible mechanisms of Shoutai Pill for the prevention and treatment of spontaneous abortion. |
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