单侧输尿管梗阻肾间质纤维化大鼠肾组织PHB1和 PHB2的表达及意义

摘要

目的：观察单侧输尿管梗阻（ UUO）肾间质纤维化（ RIF）大鼠肾组织PHB1和PHB2的表达，并探讨其意义。方法80只雄性6周龄Wistar大鼠随机分为模型组和假手术组各40只。模型组大鼠分离左侧输尿管后行双重结扎，假手术组只探查到肾包膜。于造模后第2周末和第4周末，每组各处死20只大鼠，取左侧肾组织进行肾脏病理学检查，并计算RIF指数；应用实时荧光定量PCR检测大鼠肾组织PHB1、PHB2、TGF-β1 mRNA；Western blot检测肾组织PHB1、PHB2、TGF-β1、Ⅳ型胶原（ Col-Ⅳ）和纤维连接蛋白（ FN）。结果与假手术组比较，模型组大鼠各时间点RIF指数增高，梗阻时间越长RIF指数越高，P均＜0．01；肾组织PHB1、PHB2 mRNA及蛋白表达均降低，梗阻时间越长表达水平越低，P均＜0．01；TGF-β1 mRNA及蛋白、Col-Ⅳ和FN蛋白表达均增高，P均＜0．01。肾组织PHB1蛋白表达与RIF指数、TGF-β1、Col-Ⅳ、FN呈负相关（ r分别为－0．86、－0．87、－0．70、－0．73，P均＜0．05）；PHB2蛋白表达与RIF指数、TGF-β1、Col-Ⅳ、FN呈负相关（r分别为－0．73、－0．81、－0．91、－0．84， P均＜0．05）；PHB1蛋白表达与PHB2蛋白表达呈正相关（ r＝0．78，P＜0．05）。结论在UUO所致RIF大鼠肾组织中，PHB1和PHB2表达显著降低，参与RIF的发生发展。%Objective To investigate the expression and the role of PHB1 and PHB2 in unilateral ureteral obstruction (UUO) rats with renal tubulointerstital fibrosis (RIF).Methods Eighty Wistar rats (6 week-old) were randomly divided into model group (n=40) and sham operation group (SHO) (n=40).The left ureter of rats in model group were dissocia-ted to be obstructed,while were dissociated left ureter only in SHO.At the end of the 2th and 4th weekend, 20 rats in each group were sacrificed for histological and pathological examination of left kidney.RIF index was assessed according to patho-logical results.The mRNA expression of PHB1, PHB2 and TGF-β1 were detected by RT-PCR.Western blot was performed to detect the protein expression of PHB1, PHB2, TGF-β1, collagenⅣ(Col-Ⅳ), fibronectin (FN).Results Compared to SHO group, RIF index significantly increased in each time port in model group,and the longer the obstruction, the higher the RIF index all P<0.01;significantly lower expression of PHB1 and PHB2 mRNA and protein were observed in model group, the longer the obstruction, the lower expression lever all P<0.01;significantly increased the protein expression of TGF-β1 , Col-Ⅳand FN in model group (all P<0.01).PHB1 and PHB2 protein had a negative correlation with RIF, TGF-β1, Col-Ⅳ, FN protein (all P<0.05);PHB1 protein had a positive correlation with PHB2 (P<0.05).Conclusion Low expres-sion of PHB1 and PHB2 in UUO rats with RIF, which might be involved in the progression of RIF.