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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

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、300 mg/L)作用人肾小管上皮细胞(HK?2细胞)24 h;(2)分为正常对照组、内质网应激抑制剂4?PBA(5μmol/L)组、尿酸(150 mg/L)组、4?PBA+尿酸组作用HK?2细胞24 h。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150 mg/L尿酸作用24、48、72 h HK?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、300 mg/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、225 mg/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)。48 h和72 h时150 mg/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。

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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 cells 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 cells 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+uric acid group was improved, and the protein expressions ofα?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 phenotypic transformation, and may be beneficial in attenuating uric acid?induced renal tubular damage.

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作者: 吴璞 [1] 赵菲 [2] 牛丹 [1] 王新阳 [3] 郝亚宁 [1]
期刊: 《中华肾脏病杂志》2016年32卷12期 922-927页 ISTICPKUCSCD
栏目名称: 基础研究
DOI: 10.3760/cma.j.issn.1001-7097.2016.12.007
发布时间: 2017-03-09
基金项目:
陕西省科技攻关项目(2014K-03-05-03)Fund program:Science and Technology Research Projects of Shaanxi Province
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