干扰素调节因子3在肾缺血再灌注损伤小鼠肾纤维化中的作用  

Role of interferon regulatory factor 3 in renal fibrosis in a mouse model of renal ischemia-reperfusion injury

作　　者：吴珊兰 郭晓晴 聂嘉仪 陈双全 喻文强 梁桦 Wu Shanan;Guo Xiaoqing;Nie Jiayi;Chen Shuangquan;Yu Wengqiang;Liang Hua(Department of Anesthesiology,Afiliated Foshan Women and Children Hospital,Guangdong Medical Universi-ty,Foshan528000,China)

机构地区：[1]广东医科大学附属佛山妇女儿童医院麻醉科,佛山528000

出　　处：《中华麻醉学杂志》2025年第3期353-357,共5页Chinese Journal of Anesthesiology

基　　金：广东省医学科学技术研究基金项目(B2023057);广东省自然科学基金面上项目(2024A1515012036);佛山市卫生健康局医学科研基金(20230263,20240035)。

摘　　要：目的:评价干扰素调节因子3(IRF3)在肾脏缺血再灌注损伤小鼠肾纤维化中的作用。方法:雄性C57BL/6J野生型小鼠和C57BL/6J背景巨噬细胞IRF3基因条件性敲除小鼠各12只,8~10周龄,体质量20~25 g,采用随机数字表法各分为2组(n=6):野生型假手术组(WT-Sham组)和野生型肾缺血再灌注组(WT-I/R组)、巨噬细胞IRF3基因敲除型假手术组(cKO-Sham组)和巨噬细胞IRF3基因敲除型肾缺血再灌注组(cKO-I/R组)。采用夹闭两侧肾蒂45 min后再灌注的方法制备肾缺血再灌注损伤模型。于再灌注14 d时,收集眼眶血样和肾组织,检测血清尿素(BUN)和肌酐(Cr)浓度;采用苏木精-伊红染色和天狼星红染色观察肾组织病理学结果,行肾损伤评分,测量肾纤维化面积;免疫荧光法检测肾组织纤连蛋白、Ⅰ型胶原蛋白(COL-Ⅰ)、α-平滑肌肌动蛋白(α-SMA)表达,并行F4/80-α-SMA双阳性细胞计数;荧光定量PCR检测肾组织白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和转化生长因子β1(TGF-β1)的mRNA表达。结果:2种类型小鼠,与Sham组比较,I/R组血清BUN和Cr浓度、肾纤维化面积、肾损伤评分升高,肾组织纤连蛋白、COL-Ⅰ、α-SMA表达和IL-6、IL-1β、TNF-α、TGF-β1的mRNA表达均上调,F4/80-α-SMA双阳性细胞计数升高(P<0.05)。与WT-I/R组比较,cKO-I/R组血清BUN和Cr浓度、肾纤维化面积、肾损伤评分降低,肾组织纤连蛋白、COL-Ⅰ、α-SMA表达和IL-6、IL-1β、TNF-α和TGF-β1的mRNA表达均下调,F4/80-α-SMA双阳性细胞计数降低(P<0.05)。结论:IRF3参与了肾缺血再灌注损伤小鼠肾纤维化的过程,其机制可能与促进炎症反应和巨噬细胞向肌成纤维细胞转化有关。Objective:To evaluate the role of interferon regulatory factor 3(IRF3)in renal fibrosis in a mouse model of renal ischemia-reperfusion injury.Methods:Twelve male C57BL/6J wild-type mice and 12 macrophage IRF3 conditional knockout C57BL/6J mice,aged 8-10 weeks,weighing 20-25 g,were divided into 2 groups(n=6 each)using a random number table method:wild-type sham operation group(WT-Sham group)and wild-type renal ischemia-reperfusion injury group(WT-I/R group);IRF3 conditional knockout sham operation group(cKO-Sham group)and IRF3 conditional knockout I/R group(cKO-I/R group).The model of renal I/R injury was established by occluding bilateral renal pedicles for 45 min followed by reperfusion in anesthetized animals.The orbital blood samples and renal tissues were collected at 14 days of reperfusion for determination of the concentrations of blood urea nitrogen(BUN)and creatinine(Cr)in serum,expression of fibronectin,collagen-I,alpha-smooth muscle actin(α-SMA)(by immunofluorescence),F4/80-α-SMA double positive cell count,and mRNA expression of interleukin-6(IL-6),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and transforming growth factor-β1(TGF-β1)in renal tissues(by real-time polymerase chain reaction)and for observation of pathological changes which were scored.The area of renal fibrosis was measured.Results:For two types of mice,compared with group Sham,the levels of serum BUN and Cr,area of renal fibrosis and renal injury score were significantly increased,the expression of fibronectin,COL-Ⅰandα-SMA protein and IL-6,IL-1β,TNF-αand TGF-β1 mRNA was up-regulated,and the F4/80-α-SMA dual positive cell count was increased in group WT-I/R(P<0.05).Compared with group WT-I/R,the concentrations of BUN and Cr in serum,area of renal fibrosis and renal injury score were significantly decreased,the expression of fibronectin,COL-Ⅰandα-SMA protein and IL-6,IL-1β,TNF-αand TGF-β1 mRNA was down-regulated,and the F4/80-α-SMA dual positive cell count was decreased in cKO-I/R group(P<0.05).Conclusions:IRF3

关 键 词：干扰素调节因子3 再灌注损伤 肾 肾纤维化 

分 类 号：R692[医药卫生—泌尿科学]