内质网应激参与尿酸诱导的肾小管上皮细胞表型转化  

Endoplasmic reticulum stress participates in uric acid- induced phenotypic transformation of renal tubular epithelial cells

作　　者：吴璞[1] 赵菲[2] 牛丹[1] 王新阳[3] 郝亚宁[1] 

机构地区：[1]西安交通大学第一附属医院肾内科,710061 [2]西安交通大学第一附属医院妇产科,710061 [3]西安交通大学第一附属医院泌尿外科实验室,710061

出　　处：《中华肾脏病杂志》2016年第12期922-927,共6页Chinese Journal of Nephrology

基　　金：陕西省科技攻关项目（2014K-03-05-03）

摘　　要：目的探讨内质网应激在尿酸诱导的肾小管上皮细胞表型转化过程中的作用。方法（1）不同浓度尿酸（0、75、150、225、300mg／L）作用人肾小管上皮细胞（HK-2细胞）24h；（2）分为正常对照组、内质网应激抑制剂4-PBA（5μmol／L）组、尿酸（150mg／L）组、4-PBA＋尿酸组作用HK．2细胞24h。倒置显微镜下观察各组HK-2细胞的形态变化。采用MTT法检测150mg／L尿酸作用24、48、72hHK-2细胞增殖情况。Western印迹检测各组细胞表型转化相关指标α-SMA、snail、vimentin和内质网应激反应指标葡萄糖调节蛋白78（GRP78）、磷酸化真核起始因子2α（p-eIF2α）的蛋白表达。结果与对照组比较，150、225、300mg／L尿酸组细胞形态由鹅卵石样转变为长梭形，呈类似纤维细胞样形态；150、225mg／L尿酸组表型转化和内质网应激反应相关指标的蛋白表达均高于对照组（均P〈0．05）。48h和72h时150mg／L尿酸组HK．2细胞增殖均低于相应对照组（均P〈0．01）。与尿酸组比较，4-PBA＋尿酸组细胞形态有所改善，且其内质网应激蛋白和表型转化蛋白表达均降低（均P〈0．05）。结论尿酸可诱导肾小管上皮细胞发生表型转化，且内质网应激反应参与其中；4-PBA可抑制尿酸诱导的内质网应激反应和表型转化，这为治疗高尿酸肾损害提供了有利的线索。Objective To explore the effect of endoplasmic reticulum stress （ER stress） in uric acid-induced phenotypic change in renal tubular epithelial cells （HK-2）. Methods （1） HK-2 cells were cultured with 0, 75, 150, 225, 300 mg/L uric acid for 24 h in vitro. （2） The cells were divided into normal control group, ER stress inhibitor 4-PBA （5 μmol/L） group, uric acid （150 mg/L） group and 4-PBA＋uric acid group for 24 h. Morphological changes of HK-2 cells were observed under inverted microscope. MTT assay was used to detect the proliferation of HK-2 ceils treated with 150 mg/L uric acid for 24, 48 and 72 h. The protein expressions of α-smooth muscle actin （α-SMA）, vimentin, snail, glucose regulated protein 78 （GRP78） and the phosphorylation of eukaryotic initiation factor 2α （p-eIF2α） in HK-2 cells were measured by Western blotting. Results Compared with the control group, HK-2 ceils in uric acid groups （150, 225, 300 mg/L） showed fibroblast-like appearance. The protein expressions of α-SMA, vimentin, snail, GRP78 and p-eIF2α in 150 mg/L and 225 mg/L uric acid groups were higher than those in the control group （all P 〈 0.05）. The proliferation of HK-2 cells in 150 mg/L uric acid group was lower than that in control group at 48 and 72 h （all P 〈 0.01）. Compared with the uric acid group, the cell morphology in 4-PBA＋urie acid group was improved, and the protein expressions of α-SMA, vimentin, snail, GRP78 and p-eIF2α were decreased （all P 〈 0.05）. Conclusions Uric acid may induce the phenotype transformation of renal tubular epithelial cell, and ER stress is involved. 4-PBA may inhibit the uric acid-induced ER stress response and phenotypie transformation, and may be beneficial in attenuating uric acid-induced renal tubular damage.

关 键 词：尿酸 细胞转分化 内质网应激 

分 类 号：R692[医药卫生—泌尿科学]