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目的:利用基因芯片研究Carboxyl terminus of Hsc70-interacting protein(CHIP)对肾间质纤维化TGF-β1/Smads信号通路的抑制性调节。方法:将18只SPF级SD大鼠中6只行unilateral ureteral occlusion(UUO)模型,6只植入弹性管脂肪垫加压梗阻,7天后再通定为reverse of unilateral ureteral occlusion(RUUO)组,6只UUO在7天处死留取左肾组织,另外6只为假手术组。6只RUUO大鼠在再通7天后处死,留取左侧肾组织,运用West-ern blotting检测肾组织CHIP的表达,免疫组化检测肾组织中α-SMA、CD68的阳性表达面积,定制芯片研究TGF-β1/Smads信号通路的基因变化。结果:UUO组TGF-β1、smad2、smad3、COL1a2、JunB基因显著上调,和假手术组比较上调超过1.5倍,肾组织中的α-SMA、CD68表达面积和假手术组比较有显著统计学意义P<0.01。而RUUO组CHIP表达量较UUO组更是显著增强P<0.01,TGF-β1、smad2、smad3、COL1a2、JunB基因显著下调,和UUO组比较下调超过1.5倍,肾组织中的α-SMA、CD68表达面积减小和UUO组比较有显著统计学意义P<0.01。结论:发现CHIP的过量表达能显著降低细胞内,TGF-β1、Smad2/3、COL1a2、JunB基因水平,Western blot结果也表明CHIP蛋白表达能明显降低Smads介导的基因转录活性,进一步的基因芯片结果显示CHIP蛋白能明显下调TGF-β1所诱导的下游基因JunB的表达。结果提示CHIP蛋白能抑制性调节TGF-β1/Smads信号通路,抑制肾纤维化。
AIM: To investigate the inhibitory effect of Carboxyl terminus of Hsc70-interacting protein (CHIP) on TGF-β1 / Smads signaling pathway in renal interstitial fibrosis. Methods: Twenty-eight SPF SD rats were subjected to unilateral ureteral occlusion (UUO). Six rabbits were implanted with elastic fat pad and obstructed with urethral obstruction. After 7 days, untreated ureteral occlusion (RUUO) Six UUO patients were sacrificed on the 7th day for left renal tissue, and the other 6 were sham-operated groups. Six RUUO rats were sacrificed 7 days after recanalization, left renal tissue was collected, and the expression of CHIP in renal tissues was detected by West-ern blotting. The positive expression area of α-SMA and CD68 in renal tissues was detected by immunohistochemistry. To study the gene changes of TGF-β1 / Smads signaling pathway. Results: The expression of TGF-β1, smad2, smad3, COL1a2 and JunB in UUO group was significantly up-regulated compared with sham-operated group, and the expression of α-SMA and CD68 in renal tissue was significantly increased compared with sham operation group P <0.01. The expression of CHIP in RUUO group was significantly higher than that in UUO group (P <0.01). The expression of CHB, TGF-β1, smad2, smad3, COL1a2 and JunB genes in RUUO group was significantly down-regulated more than 1.5 times compared with UUO group Compared with UUO group, the area of expression decreased significantly (P <0.01). CONCLUSION: Overexpression of CHIP can significantly reduce the level of intracellular TGF-β1, Smad2 / 3, COL1a2 and JunB genes. Western blot results also show that CHIP protein expression can significantly reduce Smads-mediated gene transcription activity. Further gene chips The results showed that CHIP protein could significantly down-regulate the expression of JunB induced by TGF-β1. The results suggest that CHIP protein can inhibit TGF-β1 / Smads signaling pathway and inhibit renal fibrosis.