糖基化终产物通过其受体激活NADPH氧化酶及其下游通路诱导L细胞的凋亡

发布时间：2018-07-17 21:12

【摘要】：目的:探讨糖基化终产物(AGEs)与其受体(RAGE)及其下游信号通路在肠道L细胞(GLUTag)凋亡中的作用。方法:首先将不同浓度的AGEs作用于GLUTag细胞24 h后,RT-PCR及Western blot检测各组细胞RAGE mRNA及蛋白表达水平。然后将细胞分为BSA对照组、AGEs 200μg/mL组、AGEs+siRNA-RAGE组、AGEs+apocynin组。采用Annexin V-FITC/PI检测细胞凋亡率,荧光探针检测细胞内活性氧水平,Western blot检测NADPH氧化酶亚单位p22~(phox)、p47~(phox)磷酸化水平和Bcl-2、Bax蛋白表达水平。结果:AGEs干预后GLUTag细胞RAGE mRNA及蛋白表达水平呈剂量依赖性增加。AGEs单独处理组与BSA对照组相比,细胞凋亡率显著增加,NADPH氧化酶亚单位p22~(phox)、p47~(phox)的表达增多,细胞内ROS水平升高,促凋亡蛋白Bax表达上调,抗凋亡蛋白Bcl-2表达下调。AGEs+apocynin组、AGEs+siRNA-RAGE组与AGEs单独处理组相比,细胞凋亡率显著下降,NADPH氧化酶亚单位p22~(phox)、p47~(phox)的表达减少,细胞内ROS水平下降,促凋亡蛋白Bax表达下调,抗凋亡蛋白Bcl-2表达上调。结论:AGEs作用于GLUTag,可上调细胞RAGE受体表达,激活NADPH氧化酶,使细胞内ROS生成增多,进而通过p53/Bax途径诱导GLUTag凋亡。
[Abstract]:Aim: to investigate the role of advanced glycation end product (ages) and its receptor (rage) and its downstream signaling pathway in the apoptosis of intestinal L cells (GLUTag). Methods: the expression of rage mRNA and protein was detected by RT-PCR and Western blot after different concentrations of ages were treated on GLUTag cells for 24 h. Then the cells were divided into two groups: BSA control group (ages 200 渭 g / mL) and ages siRNA-rage group (ages apocynin). Apoptosis rate was detected by Annexin V-FITC / Pi, reactive oxygen species (Ros) level was detected by fluorescence probe, phosphorylation level of NADPH oxidase subunit p22 ~ (phox) p47 ~ (phox) and expression of Bcl-2P Bax protein were detected by Western blot. Results the expression of rage mRNA and protein in GLUTag cells was increased in a dose-dependent manner. Compared with the control group, the apoptosis rate of GLUTag cells was significantly increased, and the expression of p22 ~ (phox) oxidase subunit p22 ~ (phox) p47 ~ (phox) was increased, and the intracellular Ros level was increased. The expression of pro-apoptotic protein Bax was up-regulated, and the expression of anti-apoptotic protein Bcl-2 was down-regulated. AGEs siRNA-rage group significantly decreased the expression of p22 ~ (phox) oxidase subunit p22 ~ (phox) p47 ~ (phox), and the intracellular Ros level in AGEs apocynin group. The expression of pro-apoptotic protein Bax was down-regulated and the expression of anti-apoptotic protein Bcl-2 was up-regulated. Conclusion the expression of rage receptor was up-regulated, NADPH oxidase was activated, Ros production was increased and GLUTag apoptosis was induced by p53 / Bax pathway.
【作者单位】：南方医科大学珠江医院内分泌科;
【基金】：国家自然科学基金青年基金项目(编号:81500623) 广东省科技计划项目(编号:2014A020212489)
【分类号】：R587.1

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