| 高糖后低糖过程通过ERK1/2信号途径增强H9c2心肌细胞自噬的功能 |
| 投稿时间：2016-08-12 修订日期：2016-08-22 点此下载全文 |
| 引用本文：毕亚光,王光宇,刘向东,张庆勇.高糖后低糖过程通过ERK1/2信号途径增强H9c2心肌细胞自噬的功能[J].医学研究杂志,2017,46(3):65-69 |
| DOI：
10.11969/j.issn.1673-548X.2017.03.017 |
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| 基金项目:上海市科委基金资助项目（13ZR1431500） |
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| 中文摘要:目的 研究高糖后低糖H9c2心肌细胞自噬活性的影响及ERK1/2在此过程中对自噬的调节作用。方法 用25mmol/L葡萄糖浓度的培养基培养H9c2细胞后用2.5mmol/L葡萄糖浓度培养基处理细胞2h作为低糖组（LG组），高糖组（HG组）维持25mmol/L的葡萄糖浓度，空白对照组（Con组）维持5.5mmol/L的葡萄糖浓度；低糖组加入ERK抑制剂U0126作为抑制剂组。采用MTT法检测H9c2细胞增殖能力；LDH检测试剂盒检测细胞毒性；Western blot法检测 ERK、p-ERK、LC3、Beclin-1的表达量。结果 与Con组和HG组相比，LG组细胞增殖能力降低，LDH活性增加；与Con组相比，HG组LC3-Ⅱ、Beclin-1表达量升高，p-ERK表达无显著改变；与Con组和HG组相比，LG组p-ERK、LC3-Ⅱ和Beclin-1表达显著增加；而与LG组比较，抑制剂组p-ERK明显降低，同时LC3-Ⅱ、Beclin-1表达以及细胞毒性降低，细胞活力增加。结论 ERK1/2信号途径促进了高糖后低糖过程中H9c2心肌细胞的自噬活性。 |
| 中文关键词:低糖 高糖 H9c2细胞 自噬 ERK/MAPK |
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| Effect of Low-after-high Glucose on the Activity of Autophagy in H9c2 Cells |
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| Abstract:Objective To investigate the effect of low-after-high glucose on the autophagy activity in H9c2 cells. Methods DMEM of 2.5mmol/L glucose concentration was used to replace that of 25mmol/L glucose concentration maintaining for 2h as the low glucose group (LG). The high glucose group (HG) was maintained 25mmol/L glucose concentration environment,and the control group (Con group) was maintained 5.5mmol/L glucose concentration environment all the time. In addition, ERK inhibitor U0126 was added to LG as the inhibitor group (LG+U0126). MTT assay was used to detect the proliferation of H9c2 cells; Lactate dehydrogenase assay was used to detect cytotoxicity. Western blott assay was used to detect the expression of ERK, p-ERK, LC3 and Beclin-1. Results Compared with the Con group and RG group, the cell proliferation capacity of LG group was reduced and the activity of LDH was increased which were all reversed in LG+U0126 group. The protein level of Beclin-1 and ratio of LC3Ⅱ/LC3-Ⅰ were increased one by one in the Con, HG and the LG group. Expression of p-ERK was significantly increased in LG group than other two groups, but had no obviously change in HG group and Con group. Moreover, LC3-Ⅱ/LC3-Ⅰ and Beclin-1 expressed in LG+U0126 group were decreased than that in LG group. Conclusion Low-after-high glucose increase autophagy activity in H9c2 cells by activating ERK/MAPK signal pathway, which may damage H9c2 myocardial cells. |
| keywords:Low-glucose High-glucose H9c2 cell Autophagy ERK/MAPK |
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