小鼠肾组织中SnoN蛋白表达水平与肾小管上皮细胞转分化的关系  

Relationship between expression of SnoN on renal tissue and epithelial-myofibroblast transition in mice

作　　者：王宁宁 王笑云 毛慧娟 王晓华 熊明霞 张飞飞 何东元 杨俊伟 

机构地区：[1]南京医科大学第一附属医院江苏省人民医院肾脏科,南京210029

出　　处：《肾脏病与透析肾移植杂志》2006年第1期25-29,共5页Chinese Journal of Nephrology,Dialysis & Transplantation

基　　金：国家自然科学基金(30270616和30470800);江苏省自然科学基金(BK2004160)资助

摘　　要：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 elucidate the role of co-repressor SnoN in renal epithelial-myofibroblast transition （EMT） induced by TGF-β, the changes of SnoN expression in normal or damaged renal tissues from mice were investigated. Methodology： The animal model of renal disease was induced by unilateral ureteral obstruction （UUO） in male CD-1 mice. These mice were divided into two groups, who were sham （n = 12） and UUO groups （n = 12）. At day third and 7th after operation, the content of TGF-β1 in renal tissue was tested by ELISA from each group. Meanwhile, the de novo expression of α-smooth muscle actin （α-SMA） was assessed by western blotting, and the total collagen level was measured by chemical way. Results ： Comparison to the sham group, the expression of SnoN in renal tissues was decreased in UUO mice, meanwhile the level of TGF-β1, α-SMA expression, and content of total collagen were increased respectively. By regression analyses, the degree of SnoN downegulation had close relationship with the level of TGF-β1, de novo expression of α-SMA, and accumulation of collagen. In vitro studies, we found that TGF-β1 decreased the expression of SnoN in HKC cells before the changes of α-SMA expression. Conclusion：These results suggest that SnoN play a key role in the pathogenesis of renal fibrosis, through attenuating the renal EMT.

关 键 词：肾小管-间质纤维化 SNON 转化生长因子β Α平滑肌肌动蛋白 

分 类 号：R692[医药卫生—泌尿科学]