摘要
目的探讨微小RNA(miR)-223对慢性肾小球肾炎(CGN)大鼠炎症反应、纤维化及白细胞介素-6(IL-6)/信号转导与转录激活因子3(STAT3)信号通路的影响。方法通过注射阳离子化牛血清白蛋白(C-BSA)建立CGN大鼠模型,随机分为模型组(CGN组,尾静脉注射等体积生理盐水)、NC agomir组(尾静脉注射50μg/kg NC agomir)、miR-223 agomir组(尾静脉注射50μg/kg miR-223 agomir)、miR-223 agomir+IL-6/STAT3通路激活剂组(miR-223 agomir+rhIL-6组,尾静脉注射50μg/kg miR-223 agomir和1μg/kg rhIL-6),以正常大鼠作为对照组(Control组,造模期间同期同部位注射等量生理盐水,给药期间尾静脉注射等体积生理盐水),每组各10只。检测5组大鼠肾功能指标[24h尿蛋白、血清尿素氮(BUN)、血肌酐(SCr)、胱抑素C(Cys-C)、视黄醇结合蛋白(RBP)]及肾组织炎症因子[IL-1β、肿瘤坏死因子(TNF)-α和单核细胞趋化蛋白(MCP)-1]水平并进行组间比较。采用HE染色和Masson染色观察大鼠肾组织形态学和纤维化情况,采用qRT-PCR法检测大鼠肾组织中miR-223表达水平。采用western blot检测IL-6、STAT3、p-STAT3蛋白表达水平。结果与Control组比较,CGN组大鼠肾脏组织可见明显病理损伤,肾间质纤维化面积增加,肾功能指标、肾组织炎症因子、IL-6蛋白表达水平及p-STAT3/STAT3比值均升高,肾组织miR-223 mRNA表达水平降低(P<0.05)。与CGN组比较,miR-223 agomir组大鼠肾组织病理损伤显著减轻,肾间质纤维化面积减少,肾功能指标、肾组织炎症因子、IL-6蛋白表达水平、p-STAT3/STAT3比值均降低,肾组织miR-223 mRNA表达水平升高(P<0.05)。与miR-223 agomir组比较,miR-223 agomir+rhIL-6组大鼠肾脏组织病理损伤明显加重,肾间质纤维化面积增加,肾功能指标、肾组织炎症因子、IL-6蛋白表达水平及p-STAT3/STAT3比值均升高(P<0.05)。结论miR-223过表达通过抑制IL-6/STAT3信号通路激活改善CGN大鼠炎症反应和纤维化。
Objective To investigate the effects of microRNA(miR)-223 on inflammation,fibrosis and interleukin-6(IL-6)/signal transduction and transcriptional activator 3(STAT3)signaling pathway in chronic glomerulonephritis(CGN)rats.Methods CGN rat model was established by injecting cationized bovine serum albumin(C-BSA),and randomly divided into model group(CGN group,equal volume of saline was injected into the tail vein),NC agomir group(50μg/kg NC agomir was injected into the tail vein),miR-223 agomir group(50μg/kg miR-223 agomir was injected into the tail vein),miR-223 agomir+IL-6/STAT3 pathway activator group(miR-223 agomir+rhIL-6 group,50μg/kg miR-223 agomir and 1μg/kg rhIL-6 were injected into the tail vein),and normal rats were used as control group(the same amount of normal saline was injected into the same place at the same time during the molding period,and the same volume of normal saline was injected into the tail vein during the administration period),with 10 rats in each group.The renal function indexes[24 h urinary protein,blood urea nitrogen(BUN),serum creatinine(SCr),cystatin C(Cys-C),retinol conjugated protein(RBP)]and renal tissue inflammatory factors[IL-1β,tumor necrosis factor(TNF)-αand monocyte chemoattractant protein(MCP)-1]of 5 groups were detected and compared between groups.HE staining and Masson staining were used to observe the renal histomorphology and fibrosis.The expression level of miR-223 in rat kidney tissues was detected by qRT-PCR.The expression levels of IL-6,STAT3 and p-STAT3 were detected by western blot.Results Compared with the control group,the renal tissue in CGN group showed obvious pathological damage,the area of renal interstitial fibrosis increased,renal function index,renal tissue inflammatory factors,IL-6 protein expression level and p-STAT3/STAT3 ratio were increased,while renal tissue miR-223 mRNA expression level was decreased(P<0.05).Compared with CGN group,the pathological damage of renal tissue in miR-223 agomir group was significantly reduced,the area of renal interstitial fibrosis decreased,renal function index,renal tissue inflammatory factors,IL-6 protein expression level and p-STAT3/STAT3 ratio were decreased,while renal tissue miR-223 mRNA expression level was increased(P<0.05).Compared with miR-223 agomir group,the pathological damage of renal tissue in miR-223 agomir+rhIL-6 group was significantly worse,the area of renal interstitial fibrosis increased,renal function index,renal tissue inflammatory factors,IL-6 protein expression level and p-STAT3/STAT3 ratio were increased(P<0.05).Conclusion Overexpression of miR-223 improves inflammation and fibrosis in CGN rats by inhibiting the activation of IL-6/STAT3 signaling pathway.
作者
韦泽丰
郑金花
王自强
Wei Zefeng;Zheng Jinhua;Wang Ziqiang(Department of Nephrology,the First Affiliated Hospital of Hainan Medical College,Haikou,570000,China)
出处
《临床内科杂志》
CAS
2023年第7期482-486,共5页
Journal of Clinical Internal Medicine
基金
海南省卫生健康行业科研项目(20A200532)。
