miR-532-3p靶向抑制Notch1信号通路对慢性肾脏病大鼠巨噬细胞极化的影响  

Effect of miR-532-3 p on macrophage polarization in rats with chronic kidney disease through targeted inhibition of Notch 1 signal pathway

作　　者：徐明芝[1] 安娜[1] 白亚飞[1] 陈汝满 贺纪清 王春莉 祁永慧 潘明娇 XU Mingzhi;AN Na;BAI Yafei;CHEN Ruman;HE Jiqing;WANG Chunli;QI Yonghui;PAN Mingjiao(Blood Purification Center,Hainan Provincial People's Hospital,Hainan Hospital Affiliated to Hainan Medical College,Haikou 570311,China)

机构地区：[1]海南省人民医院·海南医学院附属海南医院血液净化中心,海口570311

出　　处：《中国免疫学杂志》2025年第2期310-314,319,共6页Chinese Journal of Immunology

基　　金：海南省自然科学基金(822MS169)。

摘　　要：目的:分析miR-532-3p靶向抑制Notch1信号通路对慢性肾脏病(CKD)大鼠巨噬细胞极化的影响。方法:将75只SD大鼠分为对照组、CKD组、ago-NC组、ago-miR-532-3p组、FLI-06组,除对照组外均构建CKD模型并注射相应质粒及抑制剂,对照组与CKD组以等量生理盐水代替,ELISA测定大鼠血肌酐(Scr)、血尿素氮(BUN)、TNF-α、IL-1β、IL-10水平,HE染色与Masson染色观察大鼠肾组织病理及肾纤维化,流式细胞术检测巨噬细胞CD11c^(+)和CD206^(+)情况,qRT-PCR检测大鼠肾组织miR-532-3p表达,Western blot检测Notch1蛋白表达;双荧光素酶报告基因测定miR-532-3p与Notch1的靶向结合。结果:对照组大鼠肾小球、肾小管及上皮细胞结构完整,胞界清晰,排列整齐;CKD组大鼠肾小球上皮细胞坏死增多、系膜基质增多、肾小球硬化,炎症细胞浸润,肾纤维化程度加深;与CKD组相比,ago-miR-532-3p组、FLI-06组肾小球、肾小管损伤程度减小,炎症浸润细胞减少,肾纤维化程度减轻;与对照组相比,CKD组大鼠血清Scr、BUN、TNF-α、IL-1β、肾组织巨噬细胞CD11c^(+)比例、CD11c^(+)/CD206^(+)、Notch1蛋白表达升高,血清IL-10水平、肾组织CD206^(+)、miR-532-3p降低(P<0.05);与CKD组相比,ago-miR-532-3p组、FLI-06组大鼠血清Scr、BUN、TNF-α、IL-1β、肾组织巨噬细胞CD11c^(+)比例、CD11c^(+)/CD206^(+)、Notch1蛋白表达降低,血清IL-10水平、肾组织CD206^(+)、miR-532-3p表达升高(P<0.05);miR-532-3p与Notch1靶向结合,miR-532-3p过表达抑制Notch1蛋白表达。结论:促进miR-532-3p表达通过抑制Notch1通路保护CKD大鼠肾组织,其机制可能为调控巨噬细胞极化。Objective:To analyze effect of miR-532-3p on macrophage polarization in rats with chronic kidney disease(CKD)through targeted inhibition of Notch1 signal pathway.Methods:A total of 75 SD rats were divided into control group,CKD group,ago-NC group,ago-miR-532-3p group and FLI-06 group,except for control group,CKD models were constructed and corresponding plas‐mids and inhibitors were injected,control group and CKD group were replaced with same amount of normal saline.Serum creatinine(Scr),blood urea nitrogen(BUN),TNF-α,IL-1βand IL-10 were measured by ELISA,HE staining and Masson staining were used to observe renal tissue pathology and renal fibrosis in rats,flow cytometry was used to detect CD11c^(+)and CD206^(+)of macrophages,miR-532-3p expression in rat kidney tissue was detected by qRT-PCR,Notch1 protein was detected by Western blot;target binding of miR-532-3p to Notch1 was determined by double luciferase reporter gene.Results:Structure of glomerulus,renal tubules and epithelial cells was complete,cell boundaries were clear,and cells were arranged neatly in control group;glomerular epithelial cell necrosis,mesangial matrix,glomerulosclerosis,inflammatory cell infiltration and renal fibrosis were increased in CKD group;compared with CKD group,damage degree of glomerulus and tubules,inflammatory infiltration cells and renal fibrosis degree in ago-miR-532-3p group and FLI-06 group were reduced;compared with control group,serum Scr,BUN,TNF-α,IL-1β,proportion of CD11c^(+),CD11c^(+)/CD206^(+)and Notch1 protein expression in macrophages of renal tissue were increased,serum IL-10 level,CD206^(+)and miR-532-3p in renal tissue were decreased(P<0.05);compared with CKD group,serum Scr,BUN,TNF-α,IL-1β,proportion of CD11c^(+),CD11c^(+)/CD206^(+)and Notch1 protein expression in macrophages of renal tissue in ago-miR-532-3p group and FLI-06 group were decreased,serum IL-10 level,CD206^(+)and miR-532-3p in renal tissue were increased(P<0.05);miR-532-3p targeted Notch1,and overexpression of miR-532-3p inhibited Notch1 p

关 键 词：慢性肾脏病 巨噬细胞极化 miR-532-3p NOTCH1 

分 类 号：Q95-3[生物学—动物学]