目的 探讨和厚朴酚对心肌细胞脂毒性损伤的保护作用及其机制。方法 用棕榈酸(palmitic acid,PA)建立心肌细胞脂毒性损伤模型,和厚朴酚预处理后,四甲基偶氮唑蓝(MTT)法评估心肌细胞增殖能力;用试剂盒检测心肌细胞氧化应激水平及凋亡水平;Western blotting法评估AMP激活蛋白激酶(AMP-activated protein kinase,AMPK)通路及凋亡通路相关蛋白表达。结果 PA(0.4 mmol·L-1)刺激24、48 h,可显著降低细胞增殖能力,成功建立心肌细胞脂毒性损伤模型,高脂刺激条件设置为PA(0.4 mmol·L-1)刺激24 h;另一方面,高脂可显著增加细胞内活性氧(reactive oxygen species,ROS)水平,促进细胞凋亡,降低p-AMPK蛋白及抗凋亡蛋白Bcl-2表达水平,升高p-p70S6K、促凋亡蛋白Bax和cleaved caspase 3表达水平。和厚朴酚预处理,可以明显逆转上述变化。结论 和厚朴酚对心肌细胞脂毒性损伤有明显的保护作用,其作用机制与AMPK/p70S6K通路相关。
Abstract
OBJECTIVE To investigate the protective effect of honokiol on oxidative stress injury of cardiomyocytes induced by high fat and its correlation with AMP-activated protein kinase(AMPK) signaling pathway. METHODS The lipotoxic injury model of cardiomyocyte was established by palmitic acid (PA). The proliferation of cardiomyocyte was evaluated by MTT assay after pretreatment with honokiol. The oxidative stress and apoptotic level of cardiomyocyte were measured by kit. The expression of AMPK pathway and apoptotic pathway-related protein was evaluated by Western blotting. RESULTS PA (0.4 mmol·L-1) stimulation for 24 and 48 h could significantly reduce the cell proliferation ability, and a model of cardiac myocyte lipid toxicity injury was successfully established. The high-fat stimulation condition was set as PA (0.4 mmol·L-1) stimulation for 24 h, and could significantly increase the level of reactive oxygen species (ROS) in cells and promote cell death. The expression levels of p-AMPK protein and anti-apoptotic protein Bcl-2 were decreased, and the expression levels of p-p70S6K, apoptotic protein Bax and cleaved caspase 3 were increased. Pretreatment with honokiol could significantly reverse the above changes. CONCLUSION Honokiol has significant protective effect on lipid toxicity of cardiac myocytes, and its mechanism is related to the reversal of oxidative stress injury mediated by AMPK pathway.
关键词
和厚朴酚 /
高脂 /
心肌细胞 /
AMP激活蛋白激酶 /
p70S6K /
凋亡
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Key words
honokiol /
high fat /
cardiomyocyte /
AMP-activated protein kinase /
p70S6K /
apoptosis
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中图分类号：
R965
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参考文献
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脚注
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基金
国家自然科学基金项目资助(51703055);咸宁市科技局项目资助(2018XNKJ-4)
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