目的 研究肌肽对高糖诱导受损的大鼠心肌细胞（H9C2）的保护作用及其机制。 方法 用含50 mmol/L葡萄糖的高糖培养液建立高糖损伤模型，实验观察组给予5、15 mmol/L肌肽进行保护。MTT检测各组H9C2心肌细胞活力；ELISA测定培养液中IL-1β、IL-6和TNF-α炎症因子浓度含量；试剂盒检测细胞中SOD、MDA、AST和LDH活性；二氢乙锭测定细胞中活性氧含量；免疫印迹测定细胞中bax、bcl-2、p65和IκB-α的蛋白表达。 结果 肌肽预处理组细胞内活性氧含量下降，SOD活性升高，AST和LDH活性减弱，MDA含量降低；其培养液中炎症因子含量明显降低，细胞核中p65蛋白水平降低，胞浆中IκB-α蛋白水平升高，bax和bax/bcl-2比值降低。 结论 肌肽能够减轻高糖诱导的H9C2心肌细胞损伤，降低细胞炎症反应，其机制可能与ROS/NF-κB信号通路有关。
Abstract
Objective To study the protective effect of carnosine on H9C2 cardiomyocyte induced by high glucose and its mechanism. Methods High glucose culture medium containing 50 mmol/L glucose was used to establish a high glucose injury model, 5 and 15 mmol/L carnosine was given to protect H9C2 cells. MTT assay was used to detect the viability of H9C2 cells in each group. ELISA was used to determine the concentration of IL-1β, IL-6 and TNF-α inflammatory factors in culture medium. SOD, MDA, AST and LDH activities in cells were detected by kit. Active oxygen content in cells was determined by ethidium dihydrochloride. The expression of bax, bcl-2, p65 and IκB-α protein in cells was determined by Western-blot. Results Carnosine pretreatment could significantly increase cell viability, intracellular reactive oxygen species content decreased, SOD activity increased, AST and LDH activity decreased, MDA content decreased. Inflammatory factors in culture medium decreased significantly, p65 protein level in nucleus significantly decreased, and IκB-α protein level in cytoplasm increased，the expression of bax, the ratio of bax to bcl-2 increased and the differences was dose-dependent. Conclusions Carnosine can alleviate the injury of H9C2 myocardial cells induced by high glucose and reduce the inflammatory reaction of cells. Its mechanism may be related to ROS/NF-κB signaling pathway.
关键词
肌肽 /
H9C2心肌细胞 /
高糖 /
氧化应激 /
炎症
Key words
/
Carnosine /
H9C2 cardiomyocyte /
High glucose /
Oxidative stress /
Inflammation
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基金
辽宁省兴辽计划领军人才专项资金（XLYC1802017）
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