敲除核因子E2相关因子对肾肌成纤维细胞活化与纤维化的影响  

The knockout of nuclear factor E2 related factor and its effect on myofibroblast activation and renal fibrosis

作　　者：程水兵[1] 郭洋洋 朱恒悦 肖言一 郑士樟 戚如意 周俊雷 杨梅[3] 白永恒[2] CHENG Shuibing;GUO Yangyang;ZHU Hengyue;XIAO Yanyi;ZHENG Shizhang;QI Ruyi;ZHOU Junlei;YANG Mei;BAI Yongheng(Department of Trauma Surgery,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325015,China;Key Laboratory of Diagnosis and Treatment of Severe Hepato-Pancreatic Diseases of Zhejiang Province,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325015,China;Intensive Care Unit,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325015,China)

机构地区：[1]温州医科大学附属第一医院创伤外科,浙江温州325015 [2]温州医科大学附属第一医院浙江省胰腺肝脏危重性疾病诊治新技术研究重点实验室,浙江温州325015 [3]温州医科大学附属第一医院重症医学科,浙江温州3250151

出　　处：《温州医科大学学报》2020年第4期300-305,311,共7页Journal of Wenzhou Medical University

基　　金：国家自然科学基金资助项目(81772264);浙江省自然科学基金资助项目(LY17H050005);温州市科技计划项目(Y20170181)。

摘　　要：目的:研究敲除核因子E2相关因子(Nrf2)对肾组织中肌成纤维细胞的活化及间质纤维化的影响。方法:将实验小鼠分成Nrf2野生型假手术组、Nrf2野生型单侧输尿管梗阻(UUO)组、Nrf2敲除型假手术组、Nrf2敲除型UUO组4个小组,每组6只。生化分析仪检测血肌酐和尿素氮水平;糖原染色(PAS)观察肾组织损伤程度;Masson染色评价肾间质总胶原累积情况;免疫组织化学染色分析肌成纤维细胞标志物α-SMA、基质成分III型胶原及TGF-β1的表达;RT-q PCR检测TGF-β1 mRNA的表达。结果:与Nrf2野生型假手术组比,Nrf2野生型UUO组血肌酐和尿素氮水平明显升高(P<0.05)。Nrf2敲除型UUO组血肌酐和尿素氮水平与Nrf2野生型UUO组相比,差异无统计学意义(P>0.05)。PAS染色显示,与Nrf2野生型UUO组相比,Nrf2敲除型UUO组肾组织损伤明显加剧(P<0.01)。Masson染色显示,与Nrf2野生型UUO组相比,Nrf2敲除型UUO组肾间质总胶原累积明显增加(P<0.01)。免疫组织化学染色显示,与Nrf2野生型UUO组相比,Nrf2敲除型UUO组肾组织α-SMA和III型胶原的表达明显增加(P<0.05),而这可能与Nrf2敲除所致的TGF-β1 m RNA和蛋白表达提高有关(P<0.05)。结论:Nrf2敲除可加剧输尿管梗阻引起的肾损伤及纤维化程度,其机制可能为Nrf2敲除通过提高TGF-β1水平,进而促进肌成纤维细胞过度活化和胶原在肾间质的过度累积。Objective:To investigate the knockout of nuclear factor E2 related factor(Nrf2)and its effect on the activation of renal myofibroblasts and interstitial fibrosis.Methods:Mice were randomly divided into four groups:Nrf2-wild-type sham group,Nrf2-knock-out sham group,Nrf2-wild-type unilateral ureteral obstruction(UUO)group and Nrf2-knock-out UUO group,with 6 mice in each group.The levels of serum creatinine and urea nitrogen were determined by biochemical analyzer;Periodic acid-Schiff staining(PAS)was used to observe the degree of renal damage;Masson staining was used to evaluate the accumulation of total collagen in renal interstitium;immunohistochemical staining was used to analyze the expression levels of myofibroblastic markerα-SMA,matrix component type III collagen and transforming growth factorβ1(TGF-β1);qRT-PCR was used to detect the mRNA expression of TGF-β1.Results:Compared with the Nrf2-wild-type sham group,the serum creatinine and urea nitrogen levels in the Nrf2-wild-type UUO group were significantly increased(P<0.05).There was no significant difference in serum creatinine and urea nitrogen levels between the Nrf2-wild-type UUO group and the Nrf2-knock-out UUO group(P>0.05).PAS staining showed that renal tissue damage was significantly aggravated in the Nrf2-knock-out UUO group(P<0.01),compared with the Nrf2-wild-type UUO group;Masson staining showed that total accumulation of renal interstitial collagen in the Nrf2-knock-out UUO group was significantly increased(P<0.01),compared with the Nrf2-wild-type UUO group;immunohistochemical staining showed that the expression ofα-SMA and type III collagen in renal tissue of Nrf2-knock-out UUO group was significantly increased(P<0.05),compared with the Nrf2-wild-type UUO group,which was related to TGF-β1 mRNA and protein expression caused by Nrf2 knockout(P<0.05).Conclusion:Nrf2 knockout can aggravate the degree of renal injury and fibrosis caused by ureteral obstruction.Its mechanism may be related to Nrf2 knockout that increases TGF-β1 levels,resulting

关 键 词：输尿管梗阻 肌成纤维细胞活化 肾纤维化 核因子E2相关因子 转化生长因子-Β1 

分 类 号：R692[医药卫生—泌尿科学]