白细胞介素-10对肾小管上皮细胞转分化的影响

Inhibitory effect of IL-10 on renal tubular epithelial to mesenchymal transition in vitro

目的研究白细胞介素-10(interleukin-10,IL-10)对高糖诱导的人近端肾小管上皮细胞(HK-2细胞)转分化(epithelial to mesenchymal transition,EMT)的影响。方法将培养的HK-2细胞通过完全随机设计分为5组:①对照组:5.5 mmol/L的D-葡萄糖组;②高糖组:30 mmol/L的D-葡萄糖组;③30 mmol/L的D-葡萄糖+1 ng/ml IL-10组;④30mmol/L的D-葡萄糖+5 ng/ml IL-10组;⑤30 mmol/L的D-葡萄糖+25 ng/ml IL-10组。培养48 h后,RT-PCR法检测各组细胞CTGF mRNA的表达;免疫细胞化学法和Western blot法检测各组细胞α-SMA的表达;ELISA法检测各组细胞FN的分泌。结果在高糖中加入1、5、25 ng/ml的IL-10作用后,与高糖组相比HK-2细胞CTGF、α-SMA和FN表达减少,且均有统计学差异(P<0.05)。结论IL-10可以抑制高糖环境下的肾小管EMT。

Objective To investigate the effect of interleukin-10 （IL-10） on high glucose induced epithelial to mesenchymal transition in human renal tubular cells. Methods Human renal proximal tubular cell line HK-2 cells were divided randomly into 5 groups. Group 1 served as control, and its HK-2 cells were incubated with 5.5 mmol/L D-glucose. The cells of group 2 were cultured with 30 mmol/L D-glucose. For the cells of groups 3, 4 and 5, 1, 5 or 25 ng/ml IL-10 was added in their culture media beside the presupplement of 30 mmol/L D-glucose. In 48 h after culture, RT-PCR was used to detect the mRNA expression of connective tissue growth factor （CTGF）, immunocytochemical assay and Western blot analysis were used to measure the expression of alpha-smooth muscle actin （ α-SMA）, and ELISA was used to determine the secretion of fibronectin （FN） in the supernatant. Results Compared with group 2, CTGF, α-SMA and FN were all lower in groups 3, 4 and 5 （ P 〈 0. 05）. Conclusion IL-10 can inhibit the renal tubular epithelial to mesenchymal transition induced by high glucose concentration.