低糖后复糖对H9c2大鼠心肌细胞自噬活性的影响
发布时间：2019-04-26 11:39
【摘要】：目的·研究低糖后复糖对H9c2大鼠心肌细胞自噬活性的影响。方法·将H9c2细胞分为低糖组(LG)、复糖组(RG)、自噬抑制组[RG+氯喹(CQ)]和空白对照组(Con)。LG组用低糖培养基培养2 h;RG组用低糖培养基培养2 h后,用高糖培养基继续培养2 h;RG+CQ组在给予低糖和高糖处理前1 h加入CQ;Con组始终用高糖培养基培养。采用MTT法检测H9c2细胞增殖能力,LDH活性检测试剂盒检测细胞毒性,Western blotting检测p-m TOR、m TOR、p-AMPK、AMPK、LC3、beclin-1的表达量。结果·与LG组相比,RG组细胞增殖能力降低,LDH活性升高(均P0.05);与RG组相比,RG+CQ组细胞增殖能力进一步降低,LDH活性进一步升高(均P0.05)。Western blotting结果显示,与LG组相比,RG组p-AMPK、LC3Ⅱ/Ⅰ和beclin-1表达量增加,p-m TOR表达量降低(均P0.05);RG+CQ组较RG组LC3Ⅱ/Ⅰ和beclin-1表达进一步增加(均P0.05)。结论·H9c2细胞经低糖后复糖处理后自噬活性增强,这可能与激活AMPK/m TOR途径有关,且此时自噬活性增强对于心肌细胞具有保护作用。
[Abstract]:Objective to study the effect of hypoglycemic compound on autophagy of cardiac myocytes in H9c2 rats. Methods H9c2 cells were divided into low glucose group, (LG), complex group, (RG), autophagy inhibition group [RG chloroquine (CQ)] and blank control group (Con). LG group, cultured in low glucose medium for 2 hours. The RG group was cultured in low glucose medium for 2 h, and the high glucose medium was used in the high glucose medium for 2 h. The RG CQ group was added to the CQ;Con group 1 h before being treated with low glucose and high glucose, and the high glucose medium was used all the time. The proliferation ability of H9c2 cells was detected by MTT assay, and the expression of pAm TOR,m TOR,p-AMPK,LC3,beclin-1 was detected by cytotoxic, Western blotting assay with LDH activity assay kit. Results compared with LG group, the proliferation ability of RG group decreased and the activity of LDH increased (P0.05). Compared with RG group, the proliferation ability of, RG CQ group was further decreased and the activity of LDH was further increased (P0.05). Western blotting). Compared with LG group, the expression of LC3 鈪，
本文编号：2466035
[Abstract]:Objective to study the effect of hypoglycemic compound on autophagy of cardiac myocytes in H9c2 rats. Methods H9c2 cells were divided into low glucose group, (LG), complex group, (RG), autophagy inhibition group [RG chloroquine (CQ)] and blank control group (Con). LG group, cultured in low glucose medium for 2 hours. The RG group was cultured in low glucose medium for 2 h, and the high glucose medium was used in the high glucose medium for 2 h. The RG CQ group was added to the CQ;Con group 1 h before being treated with low glucose and high glucose, and the high glucose medium was used all the time. The proliferation ability of H9c2 cells was detected by MTT assay, and the expression of pAm TOR,m TOR,p-AMPK,LC3,beclin-1 was detected by cytotoxic, Western blotting assay with LDH activity assay kit. Results compared with LG group, the proliferation ability of RG group decreased and the activity of LDH increased (P0.05). Compared with RG group, the proliferation ability of, RG CQ group was further decreased and the activity of LDH was further increased (P0.05). Western blotting). Compared with LG group, the expression of LC3 鈪，
本文编号：2466035
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