甘松新酮对H9C2心肌细胞低氧损伤的作用及其机制

以氯化钴(CoCl₂)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:CoCl₂ (500 μmol/L)可抑制约50 %细胞生长，而Nar (50 μmol/L)预处理显著减轻了CoCl₂ (500 μmol/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CoCl₂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 Effect of Nardosinone on Hypoxic Injury of H9C2 Cardiomyocytes and Its Mechanism

The model of hypoxic injury of H9C2 cardiomyocytes was established with cobalt chloride (CoCl₂) 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 CoCl₂(500 μmol/L) inhibited the growth of H9C2 cells by about 50% while the pretreatment with Nar(50 μmol/L) significantly alleviated CoCl₂-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 CoCl₂-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.