Apelin对单侧输尿管结扎小鼠肾脏纤维化的影响研究  

Effects of Apelin on Renal Fibrosis in Unilateral Ureteral Obstruction Mice

作　　者：王丽妍[1] 刁宗礼[1] 张启东[1] 张育[1] 刘文虎[1] 

机构地区：[1]首都医科大学附属北京友谊医院肾内科,北京市100050

出　　处：《中国全科医学》2016年第9期1042-1048,共7页Chinese General Practice

基　　金：国家自然科学基金资助项目(81570660;81300607);北京市自然科学基金课题(7132091)

摘　　要：目的探讨生物活性肽Apelin对单侧输尿管结扎(UUO)小鼠肾脏纤维化的影响及相关机制,分析在肾脏纤维化时血清和肾脏局部Apelin/APJ系统的代偿性变化。方法 2014年1月选取7周龄雄性C57Bl/6j小鼠共60只,构建UUO的小鼠模型,并设立假手术组(Sham)作为对照。术后每天给小鼠腹腔注射Apelin-13(0.1μmol/kg)或等量0.9%氯化钠溶液(vehicle)。根据小鼠手术方式和术后用药情况分为4组(15只/组):Sham+vehicle组、Sham+Apelin组、UUO+vehicle组、UUO+Apelin组。术后2周将小鼠处死,取手术侧肾脏进行检测。采用Masson染色观察肾脏纤维化程度;采用实时荧光定量PCR(RT-PCR)法检测肾脏细胞外基质胶原〔纤维连接蛋白(Fibronectin)、Ⅰ型胶原(Collagen I)、Ⅲ型胶原(CollagenⅢ)〕表达水平以及肾脏Apelin、APJ表达水平;采用Western blotting法检测肾小管上皮细胞标志物上皮细胞钙黏蛋白(E-cadherin)、间充质细胞标志物α平滑肌肌动蛋白(α-SMA)表达水平以及转化生长因子β(TGF-β)/Smads信号通路的主要信号分子(TGF-β1、TGF-βR1、p-Smad2)表达水平;采用ELISA法检测小鼠血清Apelin表达水平。结果 4组小鼠肾脏纤维化程度比较,差异有统计学意义(P<0.05)。UUO+vehicle组和UUO+Apelin组小鼠肾脏纤维化程度均高于Sham+vehicle组和Sham+Apelin组(P<0.05);UUO+Apelin组小鼠肾脏纤维化程度低于UUO+vehicle组(P<0.05)。4组小鼠肾脏Fibronectin、CollagenⅠ、CollagenⅢ表达水平比较,差异均有统计学意义(P<0.05)。UUO+vehicle组和UUO+Apelin组小鼠肾脏Fibronectin、CollagenⅠ、CollagenⅢ表达水平高于Sham+vehicle组和Sham+Apelin组(P<0.05);UUO+Apelin组小鼠肾脏Fibronectin、CollagenⅠ、CollagenⅢ表达水平低于UUO+vehicle组(P<0.05)。4组小鼠肾脏E-cadherin、α-SMA表达水平比较,差异均有统计学意义(P<0.05)。UUO+vehicle组和UUO+Apelin组小鼠肾脏E-cadherin表达水平低于Sham+vehicle组和Sham+Apelin组,α-SMA表达水平高于Sham+vehiObjective To investigate the effect of biological active peptide Apelin and its mechanism and analyze the compensatory change of partial Apelin / APJ system of serum and kidney in the process of renal fibrosis in unilateral ureteral obstruction（ UUO） mice. Methods In January 2014,a total of 60 seven- week- old male C57 Bl /6j mice were selected. We established mice models of UUO and made Sham operated mice as controls. After surgery,intraperitoneal injection of Apelin- 13（ 0. 1 μmol/kg） or the same amount of 0. 9% sodium chloride solution（ vehicle） was administrated on mice everyday. The mice were divided into four experimental groups（ 15 mice / group） according to operation mode and postoperative drug administration：Sham ＋ vehicle group,Sham ＋ Apelin group,UUO ＋ vehicle group and UUO ＋ Apelin group. Two weeks after surgery,mice were killed and the operative kidneys were taken for various tests. Masson＇s trichrome staining was performed to evaluate the extent of renal fibrosis. RT-PCR analysis was performed to evaluate the expression of kidney extracellular matrix collagens,including Fibronectin,Collagen Ⅰ and Collagen Ⅲ,as well as the expression of Apelin and APJ in kidneys. Western blotting method was performed to determine the expression of E- cadherin and α- SMA, as well as the expression of TGF-β / Smads signaling molecules TGF-β1, TGF-βR1, and p- Smad2. ELISA method was performed to determine the expression of Apelin in serum. Results The 4 groups were significantly different in the degree of renal fibrosis（ P〈0. 05）. UUO ＋ vehicle group and UUO ＋ Apelin group were higher than Sham ＋ vehicle group and Sham ＋ Apelin group in the degree of renal fibrosis（ P〈0. 05）; UUO ＋ Apelin group was lower than UUO ＋ vehicle group in the degree of renal fibrosis（ P〈0. 05）. The 4 groups were significantly different in the expression of Fibronectin,CollagenⅠand Collagen Ⅲ（ P〈0. 05）. UUO ＋ vehicle group and UUO ＋ Apelin group were higher than Sh

关 键 词：纤维化 肾疾病 肾小管上皮-间充质转分化 小鼠 APELIN 

分 类 号：R692[医药卫生—泌尿科学]