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目的:通过观察正常和单侧输尿管梗阻(UUO)小鼠肾组织中转录共抑制因子SnoN表达水平的变化,探讨SnoN在肾小管上皮细胞转分化的作用。方法:采用结扎雄性CD1小鼠左侧输尿管的方法建立UUO动物模型。设假手术小鼠为正常对照组。应用酶联免疫吸附(ELISA)法检测肾组织中转化生长因子β1(TGFβ1)水平,酸水解比色法测定肾组织内胶原的含量。借助蛋白印迹技术,检测肾组织中SnoN和α平滑肌肌动蛋白(αSMA)蛋白的变化。以人肾小管上皮细胞(HKC)为体外研究对象,观察TGFβ1对其SnoN表达的直接作用。结果:与假手术组小鼠相比,UUO小鼠术后3天肾组织中SnoN蛋白的表达量即明显降低,术后7天时进一步降为对照组的16%。同时,UUO小鼠肾组织TGFβ1水平、αSMA蛋白表达量,及其胶原的含量亦随着病程的增加而显著增加。相关分析结果表明,SnoN的减少程度与肾组织内TGFβ1水平、αSMA的表达和胶原的聚积密切相关(P<0.01)。此外,HKC细胞培养实验结果表明,TGFβ1减低该细胞表达SnoN蛋白的作用先于αSMA蛋白的变化。结论:肾脏SnoN表达水平与肾纤维化密切相关,可能通过干扰TGFβ1所致的小管上皮细胞转分化作用影响肾组织的纤维化过程。
OBJECTIVE: To investigate the role of SnoN in renal tubular epithelial cell (HUVEC) transdifferentiation by observing the expression of SnoN, a transcription co-repressor, in renal tissue of normal and unilateral ureteral obstruction (UUO) mice. Methods: UUO animal model was established by ligating the left ureter of male CD1 mice. Leave sham mice as normal control group. The level of transforming growth factor β1 (TGFβ1) in renal tissue was detected by enzyme linked immunosorbent assay (ELISA), and the content of collagen in renal tissue was determined by acid hydrolysis colorimetry. The changes of SnoN and α-smooth muscle actin (αSMA) protein in renal tissues were detected by Western blotting. Human renal tubular epithelial cells (HKC) were used as in vitro study to observe the direct effect of TGFβ1 on SnoN expression. Results: Compared with the sham operation group, the expression of SnoN protein in renal tissue of UUO mice was significantly decreased 3 days after operation and further reduced to 16% of the control group 7 days after operation. At the same time, the level of TGFβ1 and the expression of αSMA protein in UUO mice kidney and its collagen content also increased significantly with the course of disease. Correlation analysis showed that the reduction of SnoN was closely related to the level of TGFβ1, the expression of αSMA and the accumulation of collagen in renal tissues (P <0.01). In addition, the results of HKC cell culture experiments showed that TGFβ1 decreased the expression of SnoN protein in this cell prior to the change of αSMA protein. Conclusion: The expression of SnoN in kidney is closely related to renal fibrosis, which may affect the renal fibrosis process by interfering with the transdifferentiation of tubular epithelial cells induced by TGFβ1.