Effect Of Yimu Suogong Granule Containing Serum On The Expression Of Apoptosis Related Proteins In Human Decidual Gland Stromal Cells During Early Pregnancy

Objective:To study the effect of Yimu Suogong Granule drug-containing serum on the proliferation and apoptosis of human decidual gland stromal cells in early pregnancy,and to explore its possible mechanism of action,provide the theoretical basis for its treatment of decidual residual bleeding after drug abortion,and promote the clinical development of traditional Chinese medicine.Methods:Yimu Suogong Granule drug-containing serum and blank serum were prepared,human decidual gland stromal cells were cultured in vitro,and the optimal concentration and time of Yimu Suogong Granule drug-containing serum were selected by cell counting kit(CCK-8)method for subsequent experiments.Human decidual gland stromal cells in early pregnancy were divided into low,medium and high dose groups of Yimu Suogong Granule medicated serum and low,medium and high dose groups of blank serum.The effect of Yimu Suogong Granule medicated serum on apoptosis of human decidual gland stromal cells in early pregnancy was detected by flow cytometry;The expression of Bcl-2,Bax,Caspase-3,Caspase-8,Caspase-9,Fas and Fasl m RNA was detected by real-time fluorescence quantitative assay(q-PCR);The expression of apoptosis-related proteins Bcl-2,Bax,Caspase-3,Caspase-8,Caspase-9,Fas and Fasl in human decidual gland stromal cells of early pregnancy was detected by Western blot(WB).Results: At the time of intervention for 12 hours,there was no significant difference in the inhibition rate between the drug-containing serum group and the blank serum group(P>0.05);At the time of intervention for 24 hours,the inhibition rate of the drug-containing serum group with different concentrations was significantly higher than that of the blank serum group,and there was statistical significance(P<0.01);After 48 hours of intervention,the inhibition rate of the drug-containing serum group with different concentrations was significantly higher than that of the blank serum group(P<0.01);The inhibition rate increased with the increase of concentration,but the 48-hour average OD was closest to 1,so 48-hour was selected as the best time of action concentration;According to the curve of 48-hour drug concentration and inhibition rate,the semi-inhibition concentration(IC50)was calculated to be 0.155,so 5%,10%and 15% were selected as the low,medium and high concentration groups in the follow-up experiment.There was no significant difference between the apoptosis rate of 5% medicated serum and 5% blank serum(P>0.05);The apoptosis rate of 10% drug-containing serum was significantly higher than that of 10% blank serum(P<0.01);The apoptosis rate of 15% drug-containing serum was significantly higher than that of 15% blank serum(P<0.01);And increased with the increase of concentration.Compared with the blank serum group with the same concentration,the expression of Bcl-2 m RNA in 5%,10%and 15% Yimu Suogong Granule drug-containing serum group decreased significantly(P<0.05);The expression of Bax,Caspase-3,Caspase-8,Caspase-9,Fas and Fas L m RNA increased significantly(P<0.05,P<0.01).With the increase of Yimu Suogong Granule serum concentration,the expression of Bax,Caspase-3,Caspase-8,Caspase-9,Fas and Fas L in human decidual gland stromal cells of early pregnancy showed a certain concentration dependence.Compared with the blank serum group with the same concentration,the expression of Bcl-2 protein in Yimu Suogong Granule drug-containing serum group decreased significantly(P<0.01);The protein expression of Bax,Caspase-8,Caspase-9,Fas and Fas L increased significantly(P<0.01,P<0.05).With the increase of Yimu Suogong Granule serum concentration,the secretion of Bax,Caspase-8,Caspase-9,Fas and Fas L by human decidual gland stromal cells in early pregnancy showed a certain concentration dependence.Conclusion: Yimu Suogong Granule can significantly inhibit the proliferation and induce apoptosis of human decidual gland stromal cells in early pregnancy,which may regulate the proliferation and apoptosis of human decidual gland stromal cells in early pregnancy through death receptor pathway and mitochondrial pathway.