tBHQ对草酸钙引起的肾小管上皮细胞氧化应激损伤的影响  

The effect of tBHQ on oxidative stress damage in renal tubular epithelial cells caused by calcium oxalate

作　　者：雷思羽 金晶 谢文华 何屹 Lei Siyu

机构地区：[1]浙江省嘉兴市第一医院,314000

出　　处：《浙江临床医学》2024年第9期1289-1292,共4页Zhejiang Clinical Medical Journal

摘　　要：目的分析叔丁基对苯二酚(tBHQ)对草酸钙引起的肾小管上皮细胞氧化应激损伤的影响,并探讨其与核因子E2相关因子2(Nrf2)/血红素氧合酶1(HO-1)通路的相关性。方法采用正常大鼠肾上皮样细胞系NRK-52E构建细胞模型,将细胞分为对照组、tBHQ组、草酸钙(CaOx)组、CaOx+tBHQ组4组。tBHQ组用tBHQ预处理6 h;CaOx组在含4 mmol/L CaOx的完全培养基中培养;CaOx+tBHQ组用tBHQ组预处理6 h,保持在含4 mmol/L CaOx的完全培养基中继续培养。24 h后使用光学显微镜观察NRK-52E细胞的形态及草酸钙晶体沉积情况,收集细胞及上清液,采用MTT、FCM检测细胞凋亡,采用相应试剂盒测定血清活性氧类物质(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)含量,采用免疫印迹法(WB)检测细胞中Nrf2、HO-1、醌氧化还原酶-1(NQO-1)蛋白的表达情况。结果CaOx组可见草酸钙晶体沉积,CaOx+tBHQ组仅有少量晶体沉积,其余两组无晶体沉积现象;与对照组相比,CaOx组细胞凋亡增加,ROS、MDA含量上升,SOD水平降低,Nrf2、HO-1、NQO-1表达下调;与CaOx组相比,CaOx+tBHQ组细胞凋亡减少,细胞内ROS、MDA含量降低,SOD水平升高,Nrf2、HO-1、NQO-1的表达上调。结论tBHQ能够保护草酸钙晶体导致的肾小管上皮细胞氧化应激损伤,是通过Nrf2/HO-1信号通路发挥其作用。Objective To analyze the effect of tert-butyl hydroquinone(tBHQ)on oxidative stress damage in renal tubular epithelial cells caused by calcium oxalate and to investigate its association with nuclear factor E2(Nrf 2)/heme oxygenase 1(HO-1)pathway.Methods The cell model was constructed using the normal rat renal epithelial-like cell line NRK-52E,and the cells were divided into four groups:control,tBHQ,(calcium oxalate)CaOx,and CaOx+tBHQ.The tBHQ group was pre-treated with tBHQ for 6 h;the CaOx group was cultured in complete medium containing 4 mmol/L CaOx and the CaOx+tBHQ group was pre-treated with the tBHQ group for 6 h and kept in complete medium containing 4 mmol/L CaOx for continuation.After 24 hours,the morphology of NRK-52E cells and calcium oxalate crystal deposition were observed by light microscope,cells and supernatants were collected,apoptosis was detected by MTT and FCM,(reactive oxygen species)ROS,malondialdehyde(MDA)and superoxide dismutase(SOD)content by corresponding kits,and expression of Nrf 2,HO-1 and quinone oxidoreductase-1(NQO-1)proteins was detected by immunoblotting(WB).Results The calcium oxalate crystal deposition was visible in the CaOx group,only a few crystals in the CaOx+tBHQ group,the other two groups had no crystal deposition;compared with the control group,apoptosis in the CaOx group increased,increased ROS,MDA content,decreased SOD levels,and Nrf 2,HO-1,and NQO-1 were down-regulated;compared with the CaOx group,the CaOx+tBHQ group reduced apoptosis,intracellular ROS,MDA,increased SOD levels,and Nrf 2,HO-1,and NQO-1.Conclusion tBHQ can protect against oxidative stress damage in renal tubular epithelial cells caused by calcium oxalate crystals and exert its role through the Nrf 2/HO-1 signaling pathway.

关 键 词：叔丁基对苯二酚 核因子E2相关因子2 草酸钙 氧化应激 

分 类 号：R692.4[医药卫生—泌尿科学]