基于PI3K/Akt/eNOS通路分析丹寿汤通过靶向miR-146a-5p调控子宫内膜异位症的作用机制  

Action Mechanism of Danshou Decoction in Regulating Endometriosis Through Targeting miR-146a-5p Based on PI3K/Akt/eNOS Pathway

作　　者：张丹焕 马素娟[1] 李玲玲[1] ZHANG Danhuan;MA Sujuan;LI Lingling(The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou Henan China 450000)

机构地区：[1]河南中医药大学第一附属医院,河南郑州450000

出　　处：《中医学报》2023年第8期1725-1731,共7页Acta Chinese Medicine

基　　金：中国中医科学院科技创新工程课题项目(C12021A02401)。

摘　　要：目的:探讨丹寿汤通过miR-146a-5p调控磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)/内皮一氧化氮合酶(endothelial nitric oxide synthase,eNOS)通路影响子宫内膜异位症的作用机制。方法:选取异位子宫内膜基质细胞ecESC,并转染miR-146a-5p inhibitor对照品和miR-146a-5p inhibitor,采用不同浓度的丹寿汤(0μmol·L^(-1)、50μmol·L^(-1)、100μmol·L^(-1)、150μmol·L^(-1)和200μmol·L^(-1))进行干预,CCK-8试剂盒检测细胞增殖;细胞划痕考察细胞迁移能力;Transwell实验考察细胞的侵袭能力;流式细胞术检测细胞凋亡;RT-qPCR测定miR-146a-5p的表达;Western blot检测p-PI3K、p-AKT、p-eNOS的蛋白表达水平。结果:丹寿汤能明显抑制ecESCs增殖、迁移和侵袭并促进凋亡且呈剂量依赖性(P<0.05)。丹寿汤可能通过上调miR-146a-5p抑制ecESCs增殖、迁移、侵袭并促进细胞凋亡(P<0.05)。丹寿汤可明显降低p-PI3K、p-AKT、p-eNOS的mRNA及蛋白表达水平(P<0.05),而miR-146a-5p inhibitor则逆转了丹寿汤对p-PI3K、p-AKT、p-eNOS mRNA及蛋白表达的影响(P<0.05)。结论:丹寿汤通过miR-146a-5p调控PI3K/Akt/eNOS通路调节子宫内膜异位症细胞增殖、侵袭、迁移及凋亡,缓解子宫内膜异位症。Objective:To explore the regulation effect mechanism of Danshou Decoction on endometriosis through phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKt)and endothelial nitric oxide synthase(eNOS)pathway by regulating miR-146a-5p.Methods:Ectopic endometrial stromal cells ecESC were selected and transfected with miR-146a-5p inhibitor control substance and miR-146a-5p inhibitor.Different concentrations of Danshou decoction(0μmol·L^(-1),50μmol·L^(-1),100μmol·L^(-1),150μmol·L^(-1)and 200μmol·L^(-1))were used to intervene.CCK-8 kit was used to detect cell proliferation,cell scratch,cell migration,cell invasion and apoptosis by flow cytometry.The expression of miR-146a-5p was detected by RT-qPCR,and the protein expression of p-PI3K,p-AKT and p-eNOS was detected by Western blot.Results:Danshou decoction could significantly inhibit the proliferation,migration and invasion of ecESCs and promote apoptosis in a dose-dependent manner(P<0.05).Danshou Decoction may inhibit the proliferation,migration,invasion and promote apoptosis of ecESCs by up-regulating miR-146a-5p(P<0.05).Danshou Decoction could significantly reduce the mRNA and protein expression of p-PI3K,p-AKT and p-eNOS,while miR-146a-5p inhibitor reversed the effect of Danshou Decoction on p-PI3K,p-AKT,p-eNOS mRNA and protein expression(P<0.05).Conclusion:Danshou Decoction regulates the proliferation,invasion,migration and apoptosis of endometriosis cells through miR-146a-5p regulation of PI3K/Akt/eNOS pathway,and relieves endometriosis.

关 键 词：丹寿汤 子宫内膜异位症 miR-146a-5p PI3K/Akt/eNOS信号通路 作用机制 

分 类 号：R285.5[医药卫生—中药学]