喹乙醇致肾脏毒性的内质网应激相关凋亡途径研究

发布时间：2018-06-14 07:45

本文选题：喹乙醇 + 内质网应激　；参考：《卫生研究》2015年03期

【摘要】：目的通过喹乙醇染毒人源肾小管上皮细胞(HK-2细胞)并检测活性氧(ROS)和凋亡相关蛋白的表达,探讨喹乙醇肾脏毒性产生过程及其可能的内质网应激相关凋亡机制。方法分别以不同浓度(1、2、3、4、5、6、7和8μmol/ml)喹乙醇染毒HK-2细胞24 h,噻唑蓝(MTT)比色法检测细胞增殖率以确定剂量-效应关系;Hoechst-33258荧光染色检测各组细胞凋亡形态,流式细胞仪检测各组细胞凋亡率和细胞内活性氧含量;免疫印迹法(Western blot)检测内质网应激相关凋亡蛋白葡萄糖调节蛋白78(GRP78)、葡萄糖调节蛋白94(GRP94)和凋亡促进因子CCAAT增强子结合蛋白同源蛋白(CHOP)的表达。结果根据MTT毒性实验结果 ,喹乙醇对HK-2细胞凋亡效应的适宜剂量确定为1、2、3和4μmol/ml。在喹乙醇不同剂量观察组中,喹乙醇染毒2μmol/ml以上剂量组,细胞凋亡率、内质网凋亡相关蛋白GRP79、GRP94和CHOP表达增加,喹乙醇各染毒剂量均可见活性氧水平增加(P0.05);在喹乙醇不同染毒时间观察组中,喹乙醇染毒12和24 h组,细胞凋亡率、内质网凋亡相关蛋白GRP79、GRP94表达增加,喹乙醇染毒6、12和24 h组,活性氧水平、内质网凋亡相关蛋白CHOP表达增加(P0.05)。结论喹乙醇可致肾小管上皮细胞发生细胞凋亡从而造成肾脏毒性,凋亡的发生可能与内质网应激相关凋亡途径相关。
[Abstract]:Objective to study the expression of reactive oxygen species (ROS) and apoptosis related protein in human renal tubular epithelial cells (HK-2 cells) and to explore the mechanism of renal toxicity induced by olq and its possible mechanism of apoptosis related to endoplasmic reticulum stress. Methods HK-2 cells with different concentrations (1,2,3,4,5,6,7 and 8 mol/ml) were infected with 24 h, thiazole, respectively. Blue (MTT) colorimetric assay was used to determine the cell proliferation rate to determine the dose effect relationship; Hoechst-33258 fluorescence staining was used to detect the apoptosis morphology of each group. Flow cytometry was used to detect the apoptosis rate and intracellular reactive oxygen content of each group; Western blot (Western blot) was used to detect the endoplasmic reticulum associated apoptosis protein glucose regulator protein 78 (GRP78), and the grape The expression of sugar regulated protein 94 (GRP94) and apoptosis promoting factor CCAAT enhancer binding protein homologous protein (CHOP). Results according to the results of MTT toxicity test, the optimum dosage of ethanol for the apoptosis of HK-2 cells was determined to be 1,2,3 and 4 mol/ml. in the observation group of different doses of oloethanol. The dose group over 2 mu mol/ml by olq, and the cell apoptosis Rate, the expression of GRP79, GRP94 and CHOP of endoplasmic reticulum increased, and the level of reactive oxygen species increased (P0.05) in all doses of ethyl alcohol. In the group of 12 and 24 h, the rate of apoptosis, the apoptosis related to the endoplasmic reticulum, the expression of egg white GRP79, the expression of GRP94, and the activity of 6,12 and 24 h in ethyl alcohol. The expression of apoptosis related protein CHOP in endoplasmic reticulum increased (P0.05). Conclusion ethanol can induce apoptosis in renal tubular epithelial cells and induce renal toxicity. Apoptosis may be related to the apoptosis pathway associated with endoplasmic reticulum stress.
【作者单位】：中国疾病预防控制中心职业卫生与中毒控制所毒理室中国疾病预防控制中心化学污染物与健康重点实验室;
【基金】：环保公益性行业科研专项(No.201109038)
【分类号】：R114

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