减味寿胎丸抑制miR-126-3p上调滋养细胞中FOXO3表达的药效机制

Mechanism of Jianwei Shoutai Pills in inhibiting miR-126-3p up-regulating FOXO3 expression in trophoblast cells

目的:探讨减味寿胎丸对miR-126-3p调控靶基因叉头框转录因子O3(FOXO3)在滋养细胞中表达的药效机制。方法:构建慢病毒过表达miR-126-3p滋养细胞稳转株;实时荧光定量PCR法检测慢病毒过表达miR-126-3p细胞株中FOXO3、PR mRNA表达;Western blot法检测miR-126-3p对FOXO3、PR蛋白表达的调控。双荧光素酶报告验证miR-126-3p与FOXO3、FOXO3与PR的直接结合位点。结果:与阴性对照组比较,减味寿胎丸干预慢病毒过表达miR-126-3p细胞后FOXO3、PR的mRNA、蛋白表达显著上调(P<0.05,P<0.01)。结论:减味寿胎丸可以抑制miR-126-3p的表达,上调靶基因FOXO3,进而影响PR表达,达到防治自然流产的药效作用。

Objective: To explore the pharmacodynamic mechanism of Jianwei Shoutai Pills on the expression of miR-126-3 p regulatory target gene forkhead box transcription factor O3(FOXO3) in trophoblast cells. Methods: Construct a stable trophoblast cell strain overexpressing miR-126-3 p with lentivirus;RT-PCR method to detect the expression of FOXO3 and PR mRNA in the cell line overexpressing miR-126-3 p with lentivirus;Western blot method to detect miR-126-3 p regulates FOXO3 and PR proteins. The dual luciferase report verifies the direct binding sites of miR-126-3 p and FOXO3, and FOXO3 and PR.Results: Compared with the control group, the mRNA and protein expressions of FOXO3 and PR were up-regulated after Jianwei Shoutai Pills interfered with lentivirus over-expressing miR-126-3 p cells(P<0.05, P<0.01). Conclusion: Jianwei Shoutai Pills can inhibit the expression of miR-126-3 p and up-regulate the target gene FOXO3, thereby affecting the expression of PR to achieve the efficacy of preventing spontaneous abortion.