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(P<0.05);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(P<0.05)。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。 OBJECTIVE: To explore the possible mechanism of endoplasmic reticulum stress by olaquindox-induced renal tubular epithelial cells (HK-2 cells) and the detection of reactive oxygen species (ROS) and apoptosis related proteins Apoptosis mechanism. Methods HK-2 cells were exposed to olaquindox at different concentrations (1, 2, 3, 4, 5, 6, 7 and 8 μmol / ml) for 24 h. MTT assay was used to determine the cell proliferation rate - effect; Hoechst-33258 fluorescent staining of apoptotic cells in each group were detected by flow cytometry apoptosis rate and intracellular reactive oxygen species content; Western blot detection of endoplasmic reticulum stress-related apoptosis (GRP78), glucose regulatory protein 94 (GRP94), and apoptosis promoting factor CCAAT enhancer binding protein homologous protein (CHOP). Results Based on the results of MTT toxicity test, the appropriate doses of olaquindox to HK-2 cells were determined as 1, 2, 3 and 4 μmol / ml. In the olaquindox observation group, olaquindox dose of 2μmol / ml and above, the apoptosis rate, the endoplasmic reticulum apoptosis-related protein GRP79, GRP94 and CHOP expression increased, olaquindox exposure dose can be seen in all active oxygen (P <0.05). In the olaquindox treatment groups, the apoptosis rates, the expressions of GRP79 and GRP94 in the endoplasmic reticulum increased at 12 and 24 h after olaquindox treatment, At the 6th, 12th and 24th hour, the levels of reactive oxygen species and endoplasmic reticulum apoptosis-associated protein CHOP increased (P <0.05). Conclusion Olaquindox causes renal tubular epithelial cell apoptosis and thus causes renal toxicity. The occurrence of apoptosis may be related to the apoptosis pathway related to endoplasmic reticulum stress.