蓬子菜总黄酮调节miR-205保护心肌微血管内皮细胞损伤的机制研究  

Mechanism of the protective effect of total flavonoids from Pengzicai on miR-205 protecting myocardial microvascular endothelial cells

作　　者：李怀晶[1] 全碧波[1] 尚喜雨[1] LI Huai-jing;QUAN Bi-bo;SHANG Xi-yu(Department of Biochemistry and Molecular Biology,Nanyang Medical College,Nanyang,Henan 473061,China)

机构地区：[1]南阳医学高等专科学校生物化学与分子生物学教研室,河南南阳473061

出　　处：《现代预防医学》2020年第22期4168-4172,4176,共6页Modern Preventive Medicine

基　　金：河南省医学科技攻关计划项目(2018123091)。

摘　　要：目的研究蓬子菜总黄酮(FGVL)对缺氧/复氧(H/R)诱导的心肌微血管内皮细胞损伤的影响及机制。方法实验分为对照(Con)组、缺氧/复氧组、蓬子菜总黄酮低、中、高浓度(H/R+FGVL-L、M、H)组、H/R+miR-NC组、H/R+miR-205组、H/R+FGVL+anti-miR-205组。试剂盒检测乳酸脱氢酶(LDH)水平、丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性;流式细胞术检测细胞凋亡;蛋白质印迹(western blot)法检测蛋白表达;实时荧光定量PCR(RT-qPCR)检测miR-205表达水平。结果低、中、高浓度蓬子菜总黄酮处理后,缺氧/复氧诱导的心肌微血管内皮细胞中LDH水平降低,MDA含量降低,SOD活性升高(P<0.05);细胞凋亡率降低,Bcl-2表达水平升高,Bax表达水平降低,miR-205表达水平升高,且呈浓度依赖性(P<0.05)。miR-205过表达可抑制缺氧/复氧诱导的心肌微血管内皮细胞氧化应激和凋亡。抑制miR-205表达逆转了蓬子菜总黄酮对缺氧/复氧诱导的心肌微血管内皮细胞损伤的作用。结论蓬子菜总黄酮可能通过上调miR-205表达保护缺氧/复氧诱导的心肌微血管内皮细胞损伤。Objective To investigate the effect and mechanism of Flavone from Galium Verum L(FGVL)on myocardial microvascular endothelial cell injury which is induced by hypoxia/reoxygenation(H/R).Methods The experiment was divided into control(Con)group,hypoxia/reoxygenation group,and low,medium and high concentrations of total flavonoids(H/R+FGVL-L,M,H)group,H/R+miR-NC group,H/R+miR-205 group,and H/R+FGVL+anti-miR-205 group.Kits were used to detect the lactate dehydrogenase(LDH)level,the malondialdehyde(MDA)content,the superoxide dismutase(SOD)activity.The flow cytometry was applied to detect apoptosis.The Western blot was adopted to examine protein expression,and the real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression level of miR-205.Results After the treatment of low,medium,and high concentrations of Flavone from Galium Verum L,the LDH level and the MDA content in hypoxia/reoxygenation-induced myocardial microvascular endothelial cells decreased,while the activity of SOD increased(P<0.05).Moreover,the apoptosis rate decreased,the Bcl-2 expression level increased,and Bax expression decreased.The expression of miR-205 increased,which was in a concentration-dependent manner(P<0.05).Overexpression of miR-205 could inhibit hypoxia/reoxygenation-induced oxidative stress and apoptosis of cardiac microvascular endothelial cells.Inhibiting the expression of miR-205 reversed the effect of Flavone from Galium Verum L on hypoxia/reoxygenation-induced cardiac microvascular endothelial cell injury.Conclusion Flavone from Galium Verum L may protect hypoxia/reoxygenationinduced cardiac microvascular endothelial cell injury by up-regulating the expression of miR-205.

关 键 词：蓬子菜总黄酮 miR-205 缺氧/复氧 心肌微血管内皮细胞 氧化应激 凋亡 

分 类 号：R114[医药卫生—卫生毒理学]