摘要
目的通过检测单侧输尿管梗阻(UUO)大鼠肾脏组织转化生长因子-β1(TGF-β1)和Smad3的表达,探讨TGF-β1/Smad3信号通路在肾间质纤维化(RIF)中的作用。方法雄性6周龄Wistar大鼠48只随机分为假手术组和UUO组,每组24只。UUO组大鼠麻醉后,背侧切口行左侧输尿管结扎术,假手术组大鼠在同样条件下仅探及肾包膜。分别于造模第7、14、21、28天每组各处死6只大鼠,取其左侧肾脏。HE染色和Masson染色行肾脏病理学检查,并计算RIF指数,应用Western blot法和实时荧光定量-PCR检测肾脏组织Smad3蛋白及其mRNA表达;免疫组织化学法检测肾脏组织TGF-β1、Ⅳ型胶原蛋白(Col-Ⅳ)和纤维连接蛋白(FN)的蛋白表达。结果 1.与假手术组大鼠比较,UUO组大鼠各时间点:(1)病理学检查显示,肾小管上皮细胞萎缩,胶原纤维化形成,弥散性单核细胞、淋巴细胞浸润,RIF指数显著增高(P<0.01),梗阻时间越长,RIF指数越高;(2)肾脏组织Smad3蛋白及其mRNA表达均增高(Pa<0.05),梗阻时间越长,表达水平越高;(3)肾脏组织TGF-β1、Col-Ⅳ和FN的蛋白表达均增高(Pa<0.05),梗阻时间越长,表达水平越高。2.相关性分析:(1)UUO组大鼠肾脏组织Smad3、TGF-β1蛋白表达与RIF指数均呈显著正相关(r=0.520、0.727,Pa<0.01);(2)Col-Ⅳ和FN的蛋白表达与RIF指数均呈显著正相关(r=0.794、0.786,Pa<0.01);(3)Smad3的蛋白表达与TGF-β1、Col-Ⅳ和FN的蛋白表达均呈显著正相关(r=0.592、0.672、0.508,Pa<0.01)。结论 UUO所致RIF大鼠肾脏组织TGF-β1及Smad3表达均显著升高,梗阻时间越长,表达水平越高,RIF程度越重。TGF-β1/Smad3信号通路可能参与了RIF的发生发展。
Objective To explore the expressions of transforming growth factor-β1（TGF-β1）and Smad3 after unilateral ureteral obstruction（UUO） in rats and to investigate the effects and mechanisms of TGF-β1/Smad3 signaling pathway in renal tubulointerstitial fibrosis（RIF）. Methods Forty-eight Wistar male rats（6-week-old） were randomly assigned into sham operation group（n=24） and UUO group（n=24）.The left ureter of rats in UUO group were dissociated to be obstructed after anesthesia.The rats in sham operation group were dissociated left ureter only,and not to be obstructed.Six rats in the 2 groups were killed respectively at the 7~th,14~th,21~th,28~th days after surgery,the left kidney were collected.Histological changes of renal tubular interstitium were observed by HE and Masson staining,and the index of RIF was calculated.Western blot and real-time fluorescent quantitative polymerase chain reaction（PCR） were performed to detect the protein and mRNA expression of Smad3.The expression of protein of TGF-β1,collagen-Ⅳ（Col-Ⅳ）,fibronectin（FN） were detected by immunohistochemistry method. Results 1.Compared to the sham operation group,at each same point of time of UUO group：（1）Pathologic study showed that renal tubular epithelial cell atrophy,collagen fibrosis and diffuse mononucler cells,lymphocyte infiltration in UUO group.The index of RIF in UUO group was increased remarkably（P0.01）,the longer the obstruction,the higher the RIF index;（2）The expressions of protein and mRNA of Smad3 in UUO group were all significantly increased,the longer the obstruction,the higher the expression level;（3）The expressions of protein of TGF-β1,Col-Ⅳ,FN in UUO group were all significantly increased（P_a0.05）,the longer the obstruction,the higher the expression level.2.Correlation analysis showed that：（1）the expressions of protein of Smad3 and TGF-β1 in UUO group were positive correlations with the index of RIF（r=0.520,0.727,P_a 0.01）;（2）the expressions of protein of Co
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2011年第5期325-328,共4页
Journal of Applied Clinical Pediatrics
基金
广西自然科学基金(桂科自0832121)
广西医疗卫生重点科研课题(桂卫重200917)
