The Effect Of Shashen Maidong Decoction-containing Serum Combined With Cisplatin On The Expression Of Smad3, Smad7 And TGF-β1 Protein In Human Lung Adenocarcinoma A549 Cells

Purpose:To investigate the effect of Shashen Maidong Decoction drug-containing serum combined with cisplatin on proliferation of human lung adenocarcinoma A549 cells and expression of Smad3,Smad7 and TGF-β1 protein.Material and method:Forty SD rats were randomly divided into blank serum group and 10% medium-dose Shashen Maidong Decoction drug-containing serum group.The drugcontaining serum was prepared by serum pharmacological method after being given orally once a day for 7 days.Human lung adenocarcinoma A549 cells were divided into 4 groups: blank serum group,Shashen Maidong Decoction drug-containing serum group,cisplatin group and combination group.The intervention was carried out in turn with complete culture medium,Shashen Maidong Decoction drug-containing serum(10% medium dose),cisplatin(6μg/ml),Shashen Maidong Decoction drug-containing serum combined with cisplatin(10% medium dose Shashen Maidong Decoction drug-containing serum +6μg/ml cisplatin).MTT assay was used to detect the cell proliferation inhibition rate of each experimental group acting on human lung adenocarcinoma A549 cells for 24 h,48h and 72 h,and the optimal action time was selected.Immunocytochemistry and Western Blot techniques were used to detect the effects of each experimental group on the expression of Smad3,Smad7 and TGF-β1 proteins in human lung adenocarcinoma A549 cells.Results: 1.MTT results showed that each drug group had obvious inhibition on the proliferation of human lung adenocarcinoma A549 cells,and the inhibition rate on cell proliferation was the strongest at 48 h,with statistically significant difference(P<0.05).2.Compared with the drug-containing serum group and cisplatin group of shashen ophiopogon decoction,the inhibition rate of proliferation of human lung adenocarcinoma A549 cells in the combined group is significantly higher than that in the two groups,and the difference is statistically significant(P<0.05).3.Immunocytochemistry method showed that Compared with the blank serum group,the TGF-β1 and Smad3 proteins in the drug-containing serum group,cisplatin group and combination group of Shashen Maidong Decoction have fewer brownish yellow particles in the cell plasma.In the combined group,the expression of TGF-β1 and Smad3 protein was the least in brown particles in the cell plasma.Compared with the blank serum group,Smad7 protein in the drug-containing serum group,cisplatin group and combination group expressed more brown-yellow particles in cell plasma,while Smad7 protein in combination group expressed the most brown-yellow particles in cell plasma.4.Western Blot technique was used to detect the expression of Smad3,Smad7 and TGF-β1 protein.The results showed that the expression of TGF-β1 and Smad3 protein in the drugcontaining serum group,cisplatin group and combination group was relatively lower than that in the blank serum group,with the highest reduction in the combination group,and the lowest expression of TGF-β1 and Smad3 protein with statistical significance(P <0.05).The expression of Smad7 protein in the drug-containing serum group,cisplatin group and combined group was relatively up-regulated compared with that in the blank serum group.the up-regulation was the highest in the combined group,and the expression of Smad7 protein was the highest,with statistical significance(P<0.05).Conclusion:Adenophora radix ophiopogonis decoction drug-containing serum combined with cisplatin can significantly inhibit the proliferation of human lung adenocarcinoma A549 cells,and the mechanism may be realized by up-regulating the expression of Smad7 protein and down-regulating the expression of TGF-β1 and Smad3 protein in human lung adenocarcinoma A549 cells through TGF-β/Smad signaling pathway.