小檗碱通过JNK通路调控柯萨奇病毒B3感染心肌细胞凋亡的作用

Effect of Berberine on Apoptosis of Myocardial Cells Infected by Coxsackievirus B3 Through the JNK Pathway

小檗碱是具有细胞保护作用的生物碱,能够在柯萨奇病毒B3(CVB3)感染引起的病毒性心肌炎小鼠中发挥心肌保护作用,但具体的机制未阐明。在内皮细胞中,小檗碱通过c-Jun氨基末端激酶(JNK)通路抑制细胞凋亡,因此本研究将分析小檗碱通过JNK通路调控CVB3感染心肌细胞凋亡的作用。H9c2心肌细胞分为对照组(不含药物的DMEM处理)、模型组(含CVB3的DMEM处理)、小檗碱组(含CVB3及小檗碱的DMEM处理)、小檗碱+JNK质粒组(含CVB3、小檗碱、JNK质粒的DMEM处理),检测细胞凋亡率、肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)、活性氧(ROS)、丙二醛(MDA)的含量、p-JNK、cleaved caspase-3、bax、bcl-2的表达量。结果显示,模型组的细胞凋亡率、TNF-α、IL-6、ROS、MDA的含量、p-JNK、cleaved caspase-3、bax的表达量高于对照组,bcl-2的表达量低于对照组(P<0.05);小檗碱组的细胞凋亡率、TNF-α、IL-6、ROS、MDA的含量、p-JNK、cleaved caspase-3、bax的表达量低于模型组,bcl-2的表达量高于模型组(P<0.05);小檗碱+JNK质粒组的细胞凋亡率、TNF-α、IL-6、ROS、MDA的含量、p-JNK、cleaved caspase-3、bax的表达量高于小檗碱组,bcl-2的表达量低于小檗碱组(P<0.05)。以上结果表明小檗碱对CVB3感染心肌细胞的凋亡具有抑制作用,抑制JNK通路是介导这一作用可能的分子机制。

Berberine is an alkaloid with cytoprotective effects.It can elicit myocardial protection in mice with viral myocarditis caused by coxsackievirus B3(CVB3)infection,but the exact mechanism is not known.Berberine inhibitsapoptosis of endothelial cells through the c-Jun N-terminal kinase(JNK)pathway,so we analyzed the role of berberine in regulating apoptosis of CVB3-infected cardiomyocytes through thispathway.H9c2 cardiomyocytes were divided into a control group(treatedwith Dulbecco’s modified Eagle’s medium(DMEM)without drugs),model group(treatedwith DMEM containing CVB3),berberine group(treatedwith DMEM containing CVB3 and berberine)and berberine+JNK plasmid group(treatedwith DMEM containing CVB3,berberine and JNK plasmid).Percentage of apoptotic cells,content of tumor necrosis factor(TNF)-α,interleukin(IL)-6,reactive oxygen species(ROS)and malondialdehyde(MDA),as well as expression of phosphorylated(p)-JNK,cleaved caspase-3,Bax,and B-cell lymphoma 2(bcl-2)were measured.Percentage of apoptotic cells,content of TNF-α,IL-6,ROS,and MDA,as well as expression of p-JNK,cleaved caspase-3,and Bax of the model group were higher than those of the control group,and bcl-2 expression was lower than that of the control group(P<0.05).Percentage of apoptotic cells,content of TNF-α,IL-6,ROS,and MDA,as well as expression of p-JNK,cleaved caspase-3,and Bax in the berberine group were lower than those of the model group,and bcl-2 expression was higher than that of the model group(P<0.05).Percentage of apoptotic cells,content of TNF-α,IL-6,ROS,and MDA,as well as expression of p-JNK,cleaved caspase-3,and Bax in the berberine+JNK plasmid group were higher than those of theberberine group,and bcl-2 expression was lower than that of theberberine group(P<0.05).Theseresults suggested that berberine could inhibit apoptosis of CVB3-infected cardiomyocytes,and that inhibition of JNK pathway is a possible molecular mechanism.