摘要
目的:探究姜黄素(curcumin,Cur)通过miR-34a介导的自噬途径保护心肌细胞免受高糖(high glucose,HG)诱导的损伤.方法:将心肌细胞分为Con组(正常培养细胞)、HG组(30mmol/L葡萄糖培养细胞)、HG+Cur-L组(姜黄素25 µ mol/L和葡萄糖30 mmol/L培养细胞)、HG+Cur-M组(姜黄素50 μ mol/L和葡萄糖30 mmol/L培养细胞)、HG+Cur-H组(姜黄素100 μ mol/L和葡萄糖30mmol/L培养细胞)、HG+Cur+miR-NC组(转染miR-NC,使用姜黄素100 μ mol/L和葡萄糖30mmol/L培养细胞)、HG+Cur+miR-34a(转染miR-34a,使用姜黄素100µmol/L和葡萄糖30 mmol/L培养细胞)、HG+Cu+miR-34a+3-MA组(转染miR-34a,使用姜黄素100 µ mol/L、葡萄糖30 mmol/L及自噬抑制剂3-MA10µmol/L培养细胞).采用qRT-PCR检测miR-34a表达水平;CCK8检测细胞活性;ELISA检测 LDH 含量、SOD、GSH-Px 活性;Western blot 检测 Beclin-1、LC3Ⅱ/Ⅰ、p62 蛋白表达.结果:与 Con组相比,HG组细胞活性、SOD、GSH-Px活性、p62蛋白明显降低,而LDH含量、miR-34a表达、Beclin-1、LC3Ⅱ/Ⅰ蛋白表达明显增加.与HG组比较,HG+Cur-L组、HG+Cur-M组、HG+Cur-H组细胞活性、SOD、GSH-Px活性、p62蛋白明显增加,而LDH含量、miR-34a表达、Beclin-1、LC3Ⅱ/Ⅰ蛋白表达明显降低.与 HG+Cur+miR-NC 组相比,HG+Cur+miR-34a、HG+Cur+miR-34a+3-MA 组细胞活性、SOD、GSH-Px活性、p62蛋白明显降低,而LDH含量、miR-34a表达、Beclin-1、LC3Ⅱ/Ⅰ蛋白表达明显增加.与 HG+Cur+miR-34a 组相比,HG+Cur+miR-34a+3-MA 组细胞活性、SOD、GSH-Px 活性、p62蛋白明显增加,而LDH含量、miR-34a表达、Beclin-1、LC3Ⅱ/Ⅰ蛋白表达明显降低.结论:姜黄素通过微小核糖核酸-34a介导自噬途径减缓高糖诱导的心肌细胞损伤.
Abstract
Objective:To explore how curcumin(Cur)protects cardiomyocytes from damage induced by high glucose(HG)through miR-34a-mediated autophagy pathway.Methods:Cardiomyocytes were divided into Con group(normal cultured cells),HG group(30 mmol/L glucose cultured cells),HG+Cur-L group(curcumin 25μmol/L and glucose 30 mmol/L cultured cells),HG+Cur-M group(curcumin 50 μmol/L and glucose 30 mmol/L cultured cells),HG+Cur-H group(curcumin 100 μmol/L and glucose 30 mmol/L cultured cells),HG+Cur+miR-NC group(transfected miR-NC,cultured cells with curcumin 100 μmol/L and glucose 30 mmol/L),HG+Cur+miR-34a(transfected miR-34a,used curcumin 100 μmol/L and glucose 30 mmol/L cultured cells),HG+Cur+miR-34a+3-MA group(transfected with miR-34a,using curcumin 100 μmol/L,glucose 30 mmol/L and autophagy inhibitor 3-MA 5 μmol/L cultured cells).qRT-PCR was used to detect the expression level of miR-34a;CCK8 was used to detect cell proliferation activity;ELISA was used to detect LDH content,SOD,and GSH-Px activity;Western blot was used to detect Beclin-1,LC3Ⅱ/Ⅰ,and p62 protein expression.Results:Compared with the Con group,cell viability,SOD,GSH-Px activity,and p62 protein were significantly reduced,while the LDH content,miR-34a expression,Beclin-1 and LC3Ⅱ/Ⅰ protein expression were significantly increased in the HG group.Compared with the HG group,cell viability,SOD,GSH-Px activity,and p62 protein were significantly increased,while the LDH content,miR-34a expression,Beclin-1,and LC3Ⅱ/Ⅰ protein expression were significantly decreased in the HG+Cur-L group,HG+Cur-M group and HG+Cur-H group.Compared with HG+Cur+miR-NC group,cell viability,SOD,GSH-Px activity,p62 protein were significantly decreased,while LDH content,miR-34a expression,Beclin-1 and LC3Ⅱ/Ⅰ protein expression were increased significantly in the HG+Cur+miR-34a,and HG+Cur+miR-34a+3-MA groups.Compared with the HG+Cur+miR-34a group,cell viability,SOD,GSH-Px activity,and p62 protein were significantly increased,while the LDH content,miR-34a expression,Beclin-1,LC3Ⅱ/Ⅰ protein expression were significantly reduced in the HG+Cur+miR-34a+3-MA group.Conclusions:Curcumin reduces the cardiomyocyte damage induced by HG through miR-34a-mediated autophagy pathway.
维普
万方数据