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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 effect of radiation on the expression of JLP in renal tubular epithelial cells and scaffold proteins. Methods: (1) In vitro experiments: HK-2 cells were irradiated with different doses of X-rays and cell proteins and mRNAs were extracted 24h and 48h after irradiation. JLP, α-smooth muscle actin (SMA) The expression of transforming growth factor-β1 (TGF-β1) was detected by RT-PCR. The expression of JLP mRNA was detected by RT-PCR. The changes of α-SMA after irradiation were observed by immunofluorescence. (2) In vivo experiments: JLP wild-type (jlp ~ (+ / +)) mice were divided into irradiation group and non-irradiation group. The irradiation group was irradiated with 6 Gy X-ray and the mice were killed after 7 days. Western blotting was used to detect the expression of JLP, α-SMA, TGF-β1 and COL-Ⅰ in renal tissue after irradiation. The renal pathological changes were observed by PAS staining. Results: The results of in vitro studies showed that the expression of α-SMA and TGF-β1 protein was down-regulated after irradiation, and the expression of α-SMA was increased after immunostaining. After 7 days of irradiation, jlp + / + mice showed obvious vacuolar degeneration, brush border disruption, tubular necrosis, shedding, and partial tubule regeneration. Western blotting showed that α-SMA, TGF- Increased expression. CONCLUSION: Irradiation down-regulates the expression of JLP in renal tubular epithelial cells and up-regulates the expression of TGF-β1, which further promotes the up-regulation of α-SMA and COL-Ⅰ and aggravates the renal pathological damage.