白介素-34对高糖诱导的肾小管上皮细胞转分化的影响及相关机制研究

The influence of interleukin-34 on high glucose-induced epithelial-to-mesenchymal transition in rat kidney tubular epithelial cells and underlying mechanism

目的 探索白介素-34(IL-34)对高糖(HG)诱导的肾小管上皮细胞(NRK-52E)转分化(EMT)的影响及可能机制。方法 常规培养肾小管上皮细胞,实验分为正常对照组、HG组(30 mmol/L HG作用48 h)、siRNA IL-34转染组与HG+siRNA IL-34转染组。酶联免疫方法(ELISA)检测细胞IL-34蛋白表达;相差显微镜观察各组细胞表型改变;免疫荧光及Western blot方法检测α平滑肌肌动蛋白(α-SMA)、上皮钙黏素(E-ca)、NF-κB、磷酸化的NF-κB(p-NF-κB)蛋白表达。结果 HG处理后,NRK-52E细胞的IL-34、α-SMA蛋白表达明显升高,E-ca蛋白表达降低;而siRNA IL-34降低肾小管上皮细胞α-SMA的高表达,提高E-ca蛋白的表达;同时,HG组p-NF-κB蛋白表达上调,而siRNA IL-34可下调p-NF-κB蛋白表达。结论 HG环境下IL-34促进肾小管上皮细胞的EMT,其作用机制可能是通过活化NF-κB信号通路而实现。

Objective e To investigate the effect of interleukin-34(IL-34)on high glucose(HG)-induced epithelial-to-mesenchymal transition(EMT)in rat kidney tubular epithelial cell(NRK-52E cells)and its possible mechanism.Methods Renal tubular epithelial cells were cultured routinely and divided into normal control group,HG group(treated with 30 mMHG for 48 h),siRNA IL-34 transfection group and HG+siRNA IL-34 transfection group.The expressions of IL-34 protein was detected by enzyme-linked immunosorbent assay(ELISA).Phenotypic changes were observed by phase contrast microscope.The expressions ofα-SMA,E-cadherin(E-ca),NF-kB,and p-NF-kB were detected by Western blot or immunofluorescence staining.Results After HG treatment,the protein expressions of IL-34 andα-SMA in NRK-52E cells were significantly increased,while the protein expression of E-ca was decreased,while siRNA IL-34 decreased the high expression ofα-SMA and increased the protein expression of E-ca in renal tubular epithelial cells;Meanwhile,p-NF-kB protein expression was up-regulated in HG group,while siRNA IL-34 could down-regulate p-NF-kB protein expression.Conclusion IL-34 promotes EMT in renal tubular epithelial cells in HG,and the mechanism may be related to the activation of NF-kB signaling pathway.