辐射对肾小管上皮细胞及支架蛋白JLP表达的影响

Effects of Irradiation on Renal Tubular Epithelial Cells and Scaffold Protein JLP

目的:探索辐射对肾小管上皮细胞及支架蛋白JLP表达的影响。方法:(1)体外实验:采用不同剂量X线辐照HK-2细胞,并于辐照后24h和48h提取细胞蛋白及mRNA,Western印迹法检测JLP、α-平滑肌肌动蛋白(SMA)、转长生长因子(TGF)-β1蛋白的表达;RT-PCR检测JLP mRNA的表达;免疫荧光观察辐照后α-SMA的变化。(2)体内实验:将JLP野生型(jlp^(+/+))小鼠分为辐照组和非辐照组,辐照组小鼠采用6Gy X线照射,7d后处死小鼠。Western印迹法检测肾组织辐照后JLP、α-SMA、TGF-β1、胶原蛋白(COL)-Ⅰ蛋白的表达;PAS染色观察辐照后肾脏病理改变。结果:体外研究结果显示,辐照后JLP蛋白表达下调,α-SMA、TGF-β1蛋白表达上调;免疫荧光结果显示辐照后α-SMA的荧光表达增加。jlp+/+小鼠辐照后7d肾脏病理可见明显空泡变性,刷状缘紊乱,小管坏死,脱落,部分小管再生;Western印迹结果显示,辐照后α-SMA,TGF-β1,COL-Ⅰ表达增加。结论:辐照下调肾小管上皮细胞JLP的表达,同时上调TGF-β1的表达,进一步促进α-SMA、COL-Ⅰ表达上调,加重肾脏病理损害。

Objective. To explore the effects of irradiation on renal tubular epithelial cells and scaffold protein JLP. Methods： In vitro, different doses of X-ray were used in HK-2 cells, cells protein and mRNA were collected at the end of 24 h and 48 h after irradiation. Western blot was used to detect the expression of JLP, alpha SMA, TGF-beta 1. Real-time RCR was used to assay the mRNA level of JLP. Immunofluorescence was used to observe expression of alpha SMA. In vivo, JLP wild type（JLP＋/＋ ） C57/BL6 mice were divided into irradiation group and the none irradiation group, the protein of JLP, alpha SMA and TGF-beta 1, COL-I were detected. PAS staining was used to observe the morphological changes before and after irradiation. Results： In vitro studies showed that irradiation could down-regulate JLP expression, while the expression of alpha SMA and TGF-beta 1 increased accompanied with dosage increase~ disorders of brush border, tubular necrosis and regeneration were found in JLP＋/＋ mice after irradiation. Western blot showed that irradiation could upregulate alpha SMA, TGF-beta 1,COL- I expression. Conclusion- Irradi-ation could down regulate JLP expression, and activate the TGF-betal which contributed to the expression of alpha SMA, COL- I, thus aggravating renal damage.