摘要
目的研究氧化苦参碱(OMT)对缺氧复氧大鼠心肌细胞损伤的影响和机制。方法分离培养大鼠心肌细胞,分成正常对照组(Normal组)、缺氧复氧组(H/R组)和15、30、60μmol/L OMT处理组(OMT-L组、OMT-M组、OMT-H组)以及OMT-M+脂多糖(LPS)处理组(OMT-M+LPS组)。噻唑蓝(MTT)测定增殖,试剂盒检测乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)和过氧化氢酶(catalase,CAT)水平,流式细胞术测定凋亡,Western blot检测剪切的含半胱氨酸的天冬氨酸蛋白水解酶3(C-Caspase-3)、核因子-κBp65(NF-κBp65)蛋白表达。结果与Normal组比较,H/R组细胞活力降低,细胞LDH、MDA水平升高,SOD、CAT水平降低,凋亡率和C-Caspase-3、NF-κBp65蛋白表达水平升高(P均<0.05)。与H/R组比较,OMT-L、OMT-M、OMT-H组细胞活力依次升高,细胞LDH、MDA水平依次降低,SOD、CAT水平依次升高,凋亡率和C-Caspase-3、NF-κBp65蛋白表达水平依次降低(P均<0.05)。与OMT-M组比较,OMT-M+LPS组大鼠心肌细胞活力降低,LDH和MDA水平升高,SOD和CAT活性降低,凋亡率和C-Caspase-3蛋白表达水平升高(P均<0.05)。结论氧化苦参碱通过抑制NF-κB信号通路减轻缺氧复氧大鼠心肌细胞损伤。
Objective To study the influence of oxymatrine(OMT)on cardiomyocyte injury and the mechanism in rats with hypoxia/reoxygenation(H/R).Methods Rat cardiomyocytes were isolated and cultivated,and then divided into normal group,H/R group,OMT-L group(15μmol/L),OMT-M group(30μmol/L),OMT-H group(60μmol/L)and OMT-M+LPS group.The cardiomyocyte proliferation was detected by using MTT,and levels of lactic dehydrogenase(LDH),malondialdehyde(MDA),superoxide dismutase(SOD)and catalase(CAT)were detected by using kit.The cardiomyocyte apoptosis was detected by using flow cytometry,and protein expressions of cleaved cysteinyl aspartate specific proteinase 3(C-Caspase-3)and nuclear factor-κB(NF-κB)were detected by using Western blotting assay.Results The cardiomyocyte viability decreased,levels of LDH and MDA increased,levels of SOD and CAT decreased,cardiomyocyte apoptosis rate increased and protein expressions of C-Caspase-3 and NF-κBp65 increased in H/R group compared with normal group(all P<0.05).The cardiomyocyte viability increased successively,levels of LDH and MDA decreased successively,levels of SOD and CAT increased successively,cardiomyocyte apoptosis rate and protein expressions of C-Caspase-3 and NF-κBp65 decreased successively in OMT-L group,OMT-M group and OMT-H group compared with H/R group(all P<0.05).The cardiomyocyte viability decreased,levels of LDH and MDA increased,levels of SOD and CAT decreased,cardiomyocyte apoptosis rate and protein expression of C-Caspase-3 increased in OMT-M+LPS group compared with OMT-M group(all P<0.05).Conclusion OMT can relieve cardiomyocyte injury through inhibiting NF-κB signaling pathway in H/R rats.
作者
刘婷婷
刘丽
林文果
黄大岗
Liu Tingting;Liu Li;Lin Wenguo;Huang Dagang(Department of Cardiovascular Medicine,Second People's Hospital of Yibin City,Sichuan Province,Yibin 644000,China;不详)
出处
《中国循证心血管医学杂志》
2020年第9期1129-1133,共5页
Chinese Journal of Evidence-Based Cardiovascular Medicine
