上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

低糖后复糖对H9c2大鼠心肌细胞自噬活性的影响

毕亚光， 王光宇， 刘向东， 魏盟， 张庆勇   

  1. 上海交通大学附属第六人民医院心内科，上海 200233
  • 出版日期:2016-10-28 发布日期:2016-11-29
  • 通讯作者: 张庆勇， 电子信箱： zhang_qingyong71@qq.com。
  • 作者简介:毕亚光（1989—）， 女， 硕士生； 电子信箱： 317412763@qq.com。
  • 基金资助:

    上海市自然科学基金(13ZR1431500)

Effects of glucose reperfusion after low-glucose on the activity of autophagy in rat H9c2 cells

BI Ya-guang， WANG Guang-yu， LIU Xiang-dong， WEI Meng， ZHANG Qing-yong   

  1. Department of Cardiology, Shanghai Sixth Peoples Hospital, Shanghai Jiao Tong University, Shanghai 200233
  • Online:2016-10-28 Published:2016-11-29
  • Supported by:

    Natural Science Foundation of Shanghai Municipality， 13ZR1431500

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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-mTOR、mTOR、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P<0.05）；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P<0.05）。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-mTOR表达量降低（均P<0.05）；RG+CQ组较RG组LC3Ⅱ/Ⅰ和beclin-1表达进一步增加（均P<0.05）。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/mTOR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。

关键词: 低糖, 复糖, H9c2细胞, 自噬, AMPK/mTOR

Abstract:

Objective·To investigate the effects of glucose reperfusion after low-glucose on the activity of autophagy in rat cardiomyocyte H9c2. Methods·H9c2 cells were assigned to low glucose group (LG), glucose reperfusion group (RG), autophagy inhibition group (RG+CQ), and control group (Con). LG group was treated with low glucose medium for 2 h. RG group was treated with high glucose medium for 2 h after being treated with low glucose medium for 2 h. For RG+CQ group, CQ was added 1 h before low glucose and high glucose treatment. Con group was treated with high glucose medium. MTT assay was used to detect the proliferation of H9c2 cells, lactate dehydrogenase assay was used to detect the cytotoxicity, and Western blotting was used to measure the expressions of p-mTOR, mTOR, p-AMPK, AMPK LC3, and beclin-1. Results·RG group had reduced cell proliferation capacity and increased LDH activity as compared with LG group (P<0.05). RG+CQ group had lower cell proliferation capacity and higher LDH activity as compared with RG group (P<0.05). Results of Western blotting showed that RG group had higher expressions of p-AMPK, LC3II/I, and beclin-1 and lower expression of p-mTOR as compared with LG group (P<0.05), and RG+CQ group had higher expression of LC3II/I and beclin-1 as compared with RG group (P<0.05). Conclusion·Glucose reperfusion after low-glucose increases autophagy activity in H9c2 cells. This may be related to the activation of AMPK/mTOR pathway and increased autophagy activity can protect cardiomyocytes.

Key words: low-glucose, glucose reperfusion, H9c2 cell, autophagy, AMPK/mTOR