丹参通络解毒汤含药血清调控miR-126表达对缺氧/复氧大鼠心肌微血管内皮细胞自噬的影响  

Effects of Danshen Tongluo Jiedu Decoction Medicated Serum for Regulating miR-126 Expression on Autophagy in Hypoxia/Reoxygenation Myocardial Microvascular Endothelial Cells of Rats

作　　者：颜梦凡 刘泳钊 李彩云 李鑫辉 YAN Mengfan;LIU Yongzhao;LI Caiyun;LI Xinhui(School of Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学中医学院,湖南长沙410208

出　　处：《中国中医药信息杂志》2024年第11期136-141,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82074392);湖南省自然科学基金区域联合基金(2023JJ50036);湖南省中医药管理局重点课题(A2023013);湖南省教育厅科学研究重点项目(23A0283)。

摘　　要：目的探究丹参通络解毒汤含药血清调控miR-126对缺氧/复氧大鼠心肌微血管内皮细胞(CMECs)自噬的影响。方法体外培养CMECs构建缺氧/复氧损伤细胞模型,通过慢病毒转染miR-126抑制基因,将细胞分为空白组、模型组、药物组、沉默组、空载组、沉默+药物组,试剂盒检测CMECs上清液乳酸脱氢酶(LDH)含量,流式细胞术检测细胞凋亡率,RT-PCR检测细胞miR-126 mRNA表达,Western blot检测自噬相关蛋白Atg5、Beclin1、p62表达。结果与空白组比较,模型组细胞上清液LDH含量显著升高,细胞凋亡率显著升高,miR-126 mRNA表达显著降低,Atg5、Beclin1蛋白表达显著升高,p62蛋白表达显著降低,差异均有统计学意义(P<0.01)。与模型组比较,药物组、沉默+药物组细胞上清液LDH含量显著降低,细胞凋亡率显著降低,miR-126 mRNA表达显著升高,Atg5、Beclin1蛋白表达显著降低,药物组p62蛋白表达显著升高,差异均有统计学意义(P<0.05,P<0.01);沉默组细胞上清液LDH含量、细胞凋亡率及Atg5、Beclin1蛋白表达显著升高,miR-126 mRNA表达显著降低(P<0.01)。与药物组比较,沉默组、沉默+药物组细胞上清液LDH含量和细胞凋亡率显著升高(P<0.05,P<0.01),miR-126 mRNA表达显著降低(P<0.01),沉默组Atg5、Beclin1蛋白表达显著升高(P<0.01),p62蛋白表达显著降低(P<0.01)。与沉默组比较,空载组、沉默+药物组细胞上清液LDH含量、细胞凋亡率及Atg5、Beclin1蛋白表达显著降低(P<0.01),miR-126 mRNA和p62蛋白表达显著升高(P<0.05,P<0.01)。结论丹参通络解毒汤含药血清能抑制缺氧/复氧损伤引起的CMECs自噬,抑制细胞凋亡,进而保护CMECs,其机制可能与促进miR-126表达有关。Objective To investigate the effects of Danshen Tongluo Jiedu Decoction medicated serum for regulating miR-126 on autophagy in hypoxia/reoxygenation myocardial microvascular endothelial cells(CMECs)of rats.Methods Hypoxia/reoxygenation injury cell model was constructed by culturing CMECs in vitro.The cells were divided into blank group,model group,TCM group,silencing group,empty vector group,and silencing+TCM group by lentivirus transfection with miR-126 inhibitor.Lactate dehydrogenase(LDH)content in the supernatant of CMECs was detected using a reagent kit,and cell apoptosis rate was detected using flow cytometry,the miR-126 mRNA expression was detected using RT-PCR,the expressions of autophagy-related proteins Atg5,Beclin1 and p62 proteins were detected by Western blot.Results Compared with the blank group,the LDH content in the supernatant of the model group cells significantly increased,the apoptosis rate significantly increased,the miR-126 mRNA expression significantly decreased,the Atg5 and Beclin1 protein expressions significantly increased,and the p62 protein expression significantly decreased,with statistical significance(P<0.01).Compared with the model group,the TCM group and the silencing+TCM group showed a significant decrease in LDH content in the cell supernatant,a significant decrease in cell apoptosis rate,a significant increase in miR-126 mRNA expression,a significant decrease in Atg5 and Beclin1 protein expressions,and a significant increase in p62 protein expression in the TCM group,with statistical significance(P<0.05,P<0.01);the LDH content in the supernatant,apoptosis rate,Atg5 and Beclin1 protein expressions of the silence group cells significantly increased,while miR-126 mRNA expression significantly decreased(P<0.01).Compared with the TCM group,the LDH content in the cell supernatant and apoptosis rate of the silence group and the silencing+TCM group significantly increased(P<0.05,P<0.01),while the miR-126 mRNA expression significantly decreased(P<0.01);the expressions of Atg5 and Beclin1

关 键 词：丹参通络解毒汤 MIR-126 缺氧/复氧 自噬 心肌缺血再灌注损伤 心肌微血管内皮细胞 

分 类 号：R285.5[医药卫生—中药学]