内质网应激PERK/eIF2α通路在草酸钙诱导肾小管上皮细胞损伤中的作用  

Mechanism of endoplasmic reticulum stress PERK/eIF2αpathway in calcium oxalate-induced renal tubular epithelial cell injury

作　　者：卢东[1] 蒋小雷[1] 吴明辉[1] 邓建军[1] 吴运军[1] 袁强[1] 任均碧 王永春 杨建 陈宸 Lu Dong;Jiang Xiaolei;Wu Minghui;Deng Jianjun;Wu Yunjun;Yuan Qiang;Ren Junbi;Wang Yongchun;Yang Jian;Chen Chen(Department of Urology,Sichuan 404 Hospital,Mianyang 621000,China)

机构地区：[1]四川绵阳四0四医院泌尿外科,绵阳621000

出　　处：《国际泌尿系统杂志》2022年第5期769-773,共5页International Journal of Urology and Nephrology

基　　金：绵阳市卫计委课题(201616)。

摘　　要：目的探讨内质网应激蛋白激酶R样内质网激酶(PERK)/真核翻译起始因子2α(eIF2α)通路在草酸钙诱导的肾小管上皮细胞损伤中的作用机制。方法将人肾小管上皮细胞(HK-2)分为对照组、草酸钙组和草酸钙+4-苯基丁酸组。草酸钙组和草酸钙+4-苯基丁酸组在培养基中加入终浓度为5 mmol/L的草酸钙诱导细胞损伤模型,草酸钙+4-苯基丁酸组在培养基中另加入终浓度为2 mmol/L的4-苯基丁酸。通过CCK-8法、流式细胞术和DCFH-DA荧光探针检测细胞活力、凋亡和细胞内活性氧(ROS)水平,并分析细胞内PERK/eIF2α通路中蛋白以及Caspase12和Caspase3表达水平。结果草酸钙组的OD值(0.42±0.05)低于对照组(0.71±0.08),草酸钙+4-苯基丁酸组的OD值(0.65±0.07)高于草酸钙组(均P<0.05)。草酸钙组的细胞凋亡率、ROS水平、PERK和eIF2α蛋白水平、Caspase12、Caspase3 mRNA和蛋白水平高于对照组,差异均有统计学意义(均P<0.05)。草酸钙+4-苯基丁酸组的细胞凋亡率、ROS水平、PERK和eIF2α蛋白水平、Caspase12、Caspase3的mRNA和蛋白水平低于草酸钙组,差异均有统计学意义(P<0.05)。结论在草酸钙诱导的肾小管上皮细胞损伤中使用4-苯基丁酸抑制内质网应激可抑制PERK/eIF2α通路并缓解细胞凋亡。Objective To explore the mechanism of endoplasmic reticulum stress protein kinase R-like endoplasmic reticulum kinase(PERK)/eukaryotic translation initiation factor 2α(eIF2α)pathway in calcium oxalate-induced renal tubular epithelial cell injury.Methods Human renal tubular epithelial cells HK-2 were divided into control group,calcium oxalate group and calcium oxalate+4-phenylbutyric acid group.The medium was induced by adding calcium oxalate at a final concentration of 5 mmol/L.The calcium oxalate+4-phenylbutyric acid group was added with 4-phenylbutyric acid at a final concentration of 2 mmol/L.CCK-8 method,flow cytometry and DCFH-DA fluorescent probe were used to detect cell viability,apoptosis and intracellular reactive oxygen species(ROS)levels.The expression levels of proteins in the intracellular PERK/eIF2αpathway and downstream Caspase12 and Caspase3 were analyzed.Results OD value of calcium oxalate group(0.42±0.05)was lower than that of control group(0.71±0.08),OD value of calcium oxalate+4-phenylbutyric acid group(0.65±0.07)was higher than that of calcium oxalate group(all P<0.05).The apoptosis rate,ROS level,PERK and eIF2αprotein levels,Caspase12 and Caspase3 mRNA and protein levels in calcium oxalate group were higher than those in control group,and the differences were statistically significant(all P<0.05).The apoptosis rate,ROS level,PERK and eIF2αprotein levels,mRNA and protein levels of Caspase12 and Caspase3 in calcium oxalate+4-phenylbutyrate group were lower than those in calcium oxalate group,and the differences were statistically significant(P<0.05).Conclusions Inhibition of endoplasmic reticulum stress with 4-phenylbutyrate in calcium oxalate-induced renal tubular epithelial cell injury can inhibit PERK/eIF2αpathway and alleviate cell apoptosis.

关 键 词：肾结石 草酸钙 内质网应激 上皮细胞 

分 类 号：R692.6[医药卫生—泌尿科学]