甘松新酮对H9C2心肌细胞低氧损伤的作用及其机制  

The Effect of Nardosinone on Hypoxic Injury of H9C2 Cardiomyocytes and Its Mechanism

作　　者：李红艳 赵思涵 梅显运 陈政 李旭光 孙芳云[1] LI Hongyan;ZHAO Sihan;MEI Xianyun;CHEN Zheng;LI Xuguang;SUN Fangyun(School of Medicine, Xizang Minzu University∥Key Laboratory for Basic Life Science Research of Tibet Autonomous Region, Xianyang 712082, China)

机构地区：[1]西藏民族大学医学院/西藏高原相关疾病分子遗传机制与干预研究省级重点实验室,咸阳712082

出　　处：《华南师范大学学报（自然科学版）》2021年第2期51-58,共8页Journal of South China Normal University(Natural Science Edition)

基　　金：西藏自治区科技厅项目(2015ZR-14-15);陕西省教育厅科学研究项目(18JK1179);西藏自治区高校青年教师创新支持计划项目(QCZ2016-35)。

摘　　要：以氯化钴(CoCl_(2))构建H9C2心肌细胞低氧损伤模型,研究Nar在其中的作用及机制.CCK-8试剂盒检测细胞活力,流式细胞仪测细胞凋亡,以自噬抑制剂3-MA预作用细胞探讨Nar对自噬的影响,Western Blot检测Beclin-1、LC3II/LC3I、P62、Bax、Bcl-2和Caspase-3蛋白表达,分光光度计测定乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、丙二醛(MDA)和肌酸激酶(CK)含量.结果显示:CoCl_(2)(500μmol/L)可抑制约50%细胞生长,而Nar(50μmol/L)预处理显著减轻了CoCl_(2)(500μmol/L)诱导的细胞凋亡;另外,Nar通过增加LC3II/LC3I和Beclin-1表达及促进P62降解激活了自噬,但3-MA预作用逆转了该过程;3-MA预作用同时逆转了Nar对CoCl_(2)造成的H9C2细胞凋亡和氧化损伤的保护作用.研究表明:Nar在心肌细胞低氧损伤中以诱导自噬抑制凋亡的方式保护心肌细胞,Nar可能成为治疗低氧性心脏病的潜在药物.The model of hypoxic injury of H9C2 cardiomyocytes was established with cobalt chloride(CoCl_(2))to study the role of Nardosinone(Nar)in it and its mechanism.CCK-8 kit was used to detect cell viability and flow cytometry was used to detect apoptosis.Autophagy inhibitor 3-MA was used to pretreat cells to evaluate the effect of Nar on autophagy.The expressions of Beclin-1,LC3II/LC3I,P62,Bax,Bcl-2 and Caspase-3 were detected with Western Blot,and the contents of lactate dehydrogenase(LDH),superoxide dismutase(SOD),malondialdehyde(MDA)and creatine kinase(CK)were measured with spectrophotometer.The results showed that CoCl_(2)(500μmol/L)inhibited the growth of H9C2 cells by about 50%while the pretreatment with Nar(50μmol/L)significantly alleviated CoCl_(2)-induced cell apoptosis.In addition,Nar promoted P62 degradation and increased the expressions of LC3II/LC3I and Beclin-1,which were reversed with 3-MA pretreatment.At the same time,pretreatment with 3-MA reversed the protective effects of Nar on CoCl_(2)-caused H9C2 cell apoptosis and oxidative damage.The results showed that Nar played a protective role in myocardial hypoxic injury by inducing autophagy and inhibiting apoptosis.It was suggested that Nar may be a potential drug for the treatment of hypoxic heart disease.

关 键 词：甘松新酮 自噬 凋亡 心肌细胞 低氧 氧化损伤 

分 类 号：R91[医药卫生—药学]