摘要
目的评价二甲双胍对心肌细胞缺氧复氧(H/R)损伤的作用,及Nod样受体蛋白3(NLRP3)炎性小体在其中的作用机制。方法选取原代心肌细胞随机分为对照组、H/R组、二甲双胍组、激动剂组(n=6)。对照组不作H/R处理,其他3组建立心肌细胞H/R模型。二甲双胍组和激动剂组在细胞H/R模型构建后分别加入二甲双胍(终浓度为100μmol/L)、二甲双胍联合NLRP3激动剂培养。检测各组心肌细胞活性、细胞凋亡率及乳酸脱氢酶(LDH)水平,用Western blot测定腺苷酸活化蛋白激酶(AMPK)、磷酸化AMPK(p-AMPK)、NLRP3、半胱氨酸天冬氨酸酶1(Caspase-1)、白细胞介素(IL)-1β和IL-18表达水平。结果与对照组比较,其他3组心肌细胞存活率明显降低,LDH水平、细胞凋亡率明显升高,差异有统计学意义(P<0.05)。与H/R组比较,二甲双胍组细胞存活率、心肌细胞内p-AMPK/AMPK表达明显升高,LDH水平、细胞凋亡率、NLRP3、Caspase-1和IL-1β表达明显降低(P<0.05)。激动剂组心肌细胞内NLRP3、Caspase-1和IL-1β表达明显高于二甲双胍组(3.5±1.1 vs 1.5±0.5,2.5±0.5 vs 1.5±0.2,2.5±0.5 vs 1.5±0.2,P<0.05)。结论AMPK激活剂能通过抑制心肌细胞内NLRP3炎性小体活化,减轻大鼠心肌细胞的H/R损伤。
Objective To assess the role of metformin in inducing hypoxia-reoxygenation(H/R)injury of myocardiocytes and the mechanism of NLRP3 inflammasome underlying H/R injury of myocardiocytes.Methods Primary myocardiocytes were randomly divided into control group,H/R injury group,metformin treatment group and agonist group.The primary myocardiocytes in metformin treatment group and agonist group were cultured in 100μmol/L metformin and in metformin combined with NLRP3 agonist after the H/R injury model of myocardiocytes was established for H/R injury group,met-formin treatment group and agonist group.The viability and apoptosis rate of myocardiocytes and the serum lactate dehydrogenase(LDH)level were measured.The expressions of AMPK,p-AMPK,NLRP3,Caspase-1,IL-1β and IL-18 were detected by Western blot.Results The viability of myocardiocytes was significantly lower while the serum LDH level and apoptosis rate of myocardiocytes were significantly higher in metformin treatment group and agonist group than in control group(P<0.05).The viability of myocardiocytes was significantly higher while the serum LDH level and apoptosis rate of myocardiocytes were significantly lower in metformin treatment group than in H/R injury group(P<0.05).The expression levels of p-AMPK and AMPK were significantly higher while those of NLRP3,Caspase-1 and IL-1β were significantly lower in metformin treatment group than in H/R injury group(P<0.05).The expression levels of NLRP3,Caspase-1 and IL-1β were significantly higher in agonist group than in metformin treatment group(3.5±1.1 vs 1.5±0.5,2.5±0.5 vs 1.5±0.2,2.5±0.5 vs 1.5±0.2,P<0.05).Conclusion AMPK activator can alleviate the H/R injury of myocardiocytes by inhibiting the activation of NLRP3 inflammasome in rats.
作者
李思源
邹琳
余鹏
赖晓阳
施星
华福洲
陈卓辉
张静
Li Siyuan;Zou Lin;Yu Peng;Lai Xiaoyang;Shi Xing;Hua Fuzhou;Chen Zuohui;Zhang Jing(Department of Endocrinology and Metabolism,Second Affiliated Hospital of Nanchang University,Nanchang University No.2 School of Clinical Medicine,Nanchang 330006,Jiangxi Province,China)
出处
《中华老年心脑血管病杂志》
CAS
北大核心
2021年第2期192-195,共4页
Chinese Journal of Geriatric Heart,Brain and Vessel Diseases
基金
江西省教育厅基金(180084)。
