The effects of Bushen Antai Granules on the expression of VEGF/PI3K/Akt signaling pathway in human placental microvascular endothelial cells
Objective To investigate the effect of Bushen Antai(BSAT)Granules on human placental microvascular endothelial cells(HPMVECs)based on VEGFR-2 silencing,and to explore its mechanism of action in treating recurrent spontaneous abortion(RSA)through the vascular endothelial growth factor(VEGF)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)pathway.Methods Human placental microvascular endothelial cells were constructed in vitro,and then the expression of VEGFR-2 gene was silenced by transfection of small interfering RNA(siRNA)technology.The cells were intervened with Bushen Antai Granules drug containing serum or blank serum.The human placental micro-vascular endothelial cells were divided into blank serum group(PBS+blank serum),siRNA blank serum group(PBS+VEGFR-2 siRNA+blank serum),siRNA NC blank serum group(PBS+NC siRNA+blank serum),siRNA drug serum(PBS+VEGFR-2 siRNA+BSAT drug containing serum),and drug serum group(PBS+BSAT drug containing serum)for corresponding treatment.VEGF and VEGFR-2 were detected by real-time fluorescence quantitative PCR,immunofluorescence,and Western blotting.Observe the expression of PI3K,Akt mRNA and protein,and observe whether Bushen Antai Granules can mediate angiogenesis through the PI3K/Akt pathway when VEGFR-2 gene is silenced.Results There were no significant differences in VEGF,VEGFR-2,PI3K,Akt mRNA and protein between the blank serum group and the siRNA NC blank serum group(P＞0.05);The expression of VEGF,VEGFR-2,PI3K,Akt mRNA and protein in the drug serum group was significantly higher than that in the blank serum group(P＜0.05).Compared with the blank serum group,the siRNA blank serum group showed a significant decrease in the expression of VEGF,VEGFR-2,PI3K,Akt mRNA and protein(P＜0.01).Compared with the siRNA blank serum group,the siRNA drug serum group showed a significant increase in VEGF,VEGFR-2,PI3K,and Akt(P＜0.01),and the Bushen Antai Granules could significantly increase the low expression of VEGF/PI3K/Akt.Conclusion Bushen Antai Granules activate VEGFR-2 in HPMVECs,target upregulation of PI3 K/Akt expression,mediate angiogenesis,promote maternal fetal interface vascular remodeling,and thus exert therapeutic effects on RSA.
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维普
