苦荞黄酮通过JHDM1D-AS1对缺氧/复氧心肌细胞损伤的影响  

Effect of Tartary Buckwheat Flavone on Hypoxia/Reoxygenation Cardiomyocyte Injury through JHDM1D-AS1

作　　者：李朋[1] 徐建辉 胡威[1] LI Peng;XU Jianhui;HU Wei(Xiaogan Hospital Affiliated to Wuhan University of Science and Technology,Xiaogan Central Hospital,Xiaogan 432000,Hubei,China)

机构地区：[1]武汉科技大学附属孝感医院/孝感市中心医院,湖北孝感432000

出　　处：《中西医结合心脑血管病杂志》2022年第6期991-996,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

摘　　要：目的探讨苦荞黄酮通过JHDM1D反义RNA1(JHDM1D-AS1)对缺氧/复氧心肌细胞损伤的影响及机制。方法将心肌细胞H9c2分为Con组、缺氧/复氧(H/R)组、H/R+苦荞黄酮低剂量组、H/R+苦荞黄酮中剂量组、H/R+苦荞黄酮高剂量组、H/R+pcDNA组、H/R+pcDNA-JHDM1D-AS1组、H/R+苦荞黄酮+si-NC组、H/R+苦荞黄酮+si-JHDM1D-AS1组;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;试剂盒检测超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量;实时荧光定量PCR(qRT-PCR)检测JHDM1D-AS1和miR-421的表达水平;双荧光素酶报告实验检测JHDM1D-AS1和miR-421的靶向关系。结果缺氧/复氧诱导的心肌细胞的凋亡率升高,Bax表达水平升高,Bcl-2表达水平降低,SOD活性降低,MDA水平升高,JHDM1D-AS1表达水平降低,miR-421表达水平升高,差异均有统计学意义(P<0.05);不同剂量苦荞黄酮处理后,心肌细胞凋亡率降低,Bax表达水平降低,Bcl-2表达水平升高,SOD活性升高,MDA水平降低,JHDM1D-AS1表达水平升高,miR-421表达水平降低,且呈剂量依赖性(P<0.05)。过表达JHDM1D-AS1后,心肌细胞凋亡率降低,Bax表达水平降低,Bcl-2表达水平升高,SOD活性升高,MDA水平降低,差异均有统计学意义(P<0.05)。JHDM1D-AS1靶向调控miR-421;抑制JHDM1D-AS1逆转了苦荞黄酮对缺氧/复氧心肌细胞损伤的影响。结论苦荞黄酮通过调控JHDM1D-AS1/miR-421抑制缺氧/复氧诱导的心肌细胞损伤。Objective To explore the effect and mechanism of tartary buckwheat flavone on hypoxia/reoxygenation(H/R)cardiomyocyte injury through JHDM1D antisense RNA 1(JHDM1D-AS1).Methods Cardiomyocytes H9c2 were divided into Con group,H/R group,H/R+tartary buckwheat flavone low,medium and high dose group,H/R+pcDNA group,H/R+pcDNA-JHDM1D-AS1 group,H/R+tartary buckwheat flavone+si-NC group,and H/R+tartary buckwheat flavone+si-JHDM1D-AS1 group,respectively.Flow cytometry was used to detect cell apoptosis.Western blot was used to detect protein expression.Kit was used to detect superoxide dismutase(SOD)activity and malondialdehyde(MDA)content.Real-time fluorescent quantitative PCR(qRT-PCR)was used to detect the expression levels between JHDM1D-AS1 and miR-421.Dual luciferase reporter experiment was used to detect the targeting relationship between JHDM1D-AS1 and miR-421.Results Hypoxia/reoxygenation-induced cardiomyocyte apoptosis rate was increased.Bax expression level was increased.Bcl-2 expression level was decreased.SOD activity was decreased.MDA content was increased.JHDM1D-AS1 expression level was decreased.miR-421 expression level was increased(P<0.05).After treatment with different doses of tartary buckwheat flavonoids,cardiomyocyte apoptosis rate was decreased.Bax expression level was decreased.Bcl-2 expression level was increased.SOD activity was increased.MDA content was decreased.JHDM1D-AS1 expression level was increased,the expression level of miR-421 was decreased,all of them were in a dose-dependent manner(P<0.05).After overexpression of JHDM1D-AS1,cardiomyocyte apoptosis rate was decreased,Bax expression level was decreased,Bcl-2 expression level was increased,SOD activity was increased,MDA content was decreased(P<0.05).JHDM1D-AS1 targeted and regulated miR-421.Inhibition of JHDM1D-AS1 reversed the effect of tartary buckwheat flavonoids on hypoxia-reoxygenation cardiomyocyte damage.Conclusion Tartary buckwheat flavonoids could inhibit hypoxia/reoxygenation-induced cardiomyocyte damage by regulating JHDM1D-AS1/mi

关 键 词：心肌细胞损伤 缺氧/复氧 苦荞黄酮 JHDM1D反义RNA1 实验研究 

分 类 号：R28[医药卫生—中药学]