高糖经内质网应激途径诱导胰岛内皮细胞凋亡
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国家重点基础研究发展计划(973计划)(2010CB535008)


Hyperglycemia induced islet endothelial cell line MS-1 apoptosis through endoplasmic reticulum stress pathway
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    摘要:

    目的:探讨高糖诱导胰岛微血管内皮细胞株MS-1凋亡及其可能机制-方法:体外培养MS-1细胞,用含有不同浓度葡萄糖(5.6-25.0和33.6 mmol/L)的培养液分组培养12-24 h-流式细胞术分析各组细胞凋亡率变化;四甲基偶氮唑盐(MTT)法检测各组细胞增殖率变化;RT-PCR分析GRP78-CHOP-caspase-3-caspase-12 mRNA表达变化情况;Western blot分析GRP78-CHOP蛋白表达变化情况-结果:与5.6 mmol/L组相比,培养12 h及24 h后,25.0和33.6 mmol/L组MS-1细胞凋亡率显著增加(P < 0.05);而细胞增殖率显著下降(P < 0.05),且呈浓度和时间依赖性-进一步研究表明,在高糖刺激12 h及24 h后,caspase-3-caspase-12 mRNA表达显著上调(P < 0.05),CHOP mRNA和蛋白表达水平均显著上调(P < 0.05);而GRP78 mRNA和蛋白表达水平在刺激12 h后显著上调,24 h后却显著下降(P < 0.05)-结论:高糖可以促进胰岛内皮细胞凋亡增加,并呈浓度和时间依赖性升高,其机制可能与启动胰岛内皮细胞内质网应激有关-

    Abstract:

    Objective:To investigate the effect of hyperglycemia on the apoptosis and proliferation of islet endothelial cell line MS-1. Methods: MS-1 cells were treated with different concentrations of glucose (5.6,25.0 and 33.6 mmol/L) for different time (12 or 24 h). The apoptotic cells were detected by flow cytometry,and the cell proliferation was tested using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay. The expressions of GRP78,CHOP,caspase-3,caspase-12 mRNA were examined by reverse transcription and polymerase chain reaction(RT-PCR),and the expressions of GRP78,CHOP protein were examined by Western blot. Results: After treated with glucose for 12 h or 24 h,the apoptotic rates were obviously higher in 25.0 and 33.6 mmol/L group than in 5.6 mmol/L group(P < 0.05),while the cell proliferation was significantly lower in 25.0 and 33.6 mmol/L glucose treated group than that in 5.6 mmol/L glucose treated group(P < 0.05). The mRNA levels of CHOP,caspase-3,caspase-12 and the protein level of CHOP increased in a dose-dependent manner after treated with glucose(P < 0.05);while the mRNA and protein levels of GRP78 increased after treated for 12 h,and decreased at 24 h(P < 0.05). Conclusion: Hyperglycemia significantly promotes apoptosis and decreases proliferation of MS-1 cells,the mechanism may account for endoplasmic reticulum stress.

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刘 璇,徐宽枫,陈 恒,许馨予,杨 涛.高糖经内质网应激途径诱导胰岛内皮细胞凋亡[J].南京医科大学学报(自然科学版),2012,(3):349-353

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  • 收稿日期:2011-11-12
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