肠黏膜TLR4/NF-κB通路激活与伪无菌IgA肾病小鼠肾损害有关  

Activation of intestinal mucosal TLR4/NF-κB pathway is associated with renal damage in mice with pseudo-sterile IgA nephropathy

作　　者：唐余燕 孙蔚倩[1] 贺海东[1] 胡屏[1] 金玫萍[1] 刘萍[1] 黄鲁生 徐旭东[1] TANG Yuyan;Sun Weiqian;HE Haidong;HU Ping;JIN Meiping;LIU Ping;HUANG Lusheng;XU Xudong(Department of Nephrology,Minhang Hospital,Fudan University,Shanghai 201199,China)

机构地区：[1]复旦大学附属闵行医院(上海市闵行区中心医院肾脏科),上海201199

出　　处：《细胞与分子免疫学杂志》2023年第10期865-871,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81774060);上海市科委自然科学基金(21dz1200200);上海市闵行区卫生健康委员会科研课题(2022MW13);上海市闵行区特色专科建设项目(2020MWTZB07);闵行区高层次专科骨干医师培养计划(2020MZYS19);上海市闵行区自然科学研究课题(2022MHZ004)。

摘　　要：目的探讨肠黏膜Toll样受体4/核因子κB(TLR4/NF-κB)信号通路对伪无菌IgA肾病(IgAN)小鼠肾脏损害的影响。方法C57BL/6小鼠随机分为实验组(伪无菌小鼠造模组)、对照组(正常小鼠造模组)、伪无菌小鼠空白组、正常小鼠空白组。采用四联抗生素灌胃,每天1次,连续14 d,构建伪无菌小鼠。在伪无菌小鼠的基础上,采用牛血清白蛋白(BSA)联合葡萄球菌肠毒素(SEB)建立IgAN模型,通过免疫荧光染色及过碘酸希夫(PAS)染色观察肾组织病理形态学变化,ELISA分析肠黏膜屏障损伤指标脂多糖(LPS)、可溶性细胞间黏附分子1(sICAM-1)和右旋乳酸(D-LAC)的含量,生化分析检测24 h尿蛋白、血肌酐及尿素氮,反转录PCR及Western blot法检测Toll样受体4(TLR4)、髓样分化因子88(MyD88)、核因子κB(NF-κB)mRNA及蛋白水平。结果伪无菌IgAN小鼠的肾脏损伤程度较IgAN小鼠更严重,并且伪无菌IgAN小鼠的肠道黏膜LPS、sICAM-1和D-LAC表达显著升高。此外,在IgAN伪无菌小鼠的肠组织中,TLR4、MyD88、NF-κB水平的表达均上调。结论肠道菌群紊乱导致肠黏膜屏障损伤诱导TLR4信号通路的激活介导IgAN的肾脏损伤。Objective To investigate the effect of intestinal mucosal Toll-like receptor 4/nuclear factorκB(TLR4/NF-κB)signaling pathway on renal damage in pseudo-sterile IgA nephropathy(IgAN)mice.Methods C57BL/6 mice were randomly divided into experimental group(pseudosterile mouse model group),control group(IgAN mouse model group),pseudosterile mouse blank group,and normal mouse blank group.Pseudosterile mice were established by intragastric administration of quadruple antibiotics once a day for 14 days.The pseudosterile IgAN mouse model was set up by combination of oral bovine serum albumin(BSA)administration and staphylococcal enterotoxin B(SEB)injection.The pathological changes of renal tissue were observed by immunofluorescence staining and PAS staining,and the intestinal mucosa barrier damage indicators lipopolysaccharide(LPS),soluble intercellular adhesion molecule 1(sICAM-1)and D-lactate(D-LAC)were analyzed by ELISA.Biochemical analysis was used to test 24 hour urine protein,serum creatinine and blood urea nitrogen.The mRNA and protein levels of Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88)and nuclear factorκB(NF-κB)were detected by reverse transcription PCR and Western blot analysis.Results The kidney damage of pseudosterile IgAN mice was more severe than that of IgAN mice,and the expressions of intestinal mucosal barrier damage markers(LPS,sICAM-1 and D-LAC)were significantly increased in pseudosterile IgAN mice.In addition,the expressions of TLR4,MyD88,and NF-κB level were all up-regulated in the intestinal tissues of IgAN pseudosterile mice.Conclusion Intestinal flora disturbance leads to intestinal mucosal barrier damage and induces activation of TLR4 signaling pathway to mediate renal injury in IgAN.

关 键 词：IgA肾病(IgAN) 伪无菌IgA肾病小鼠 Toll样受体4(TLR4) 核因子κB(NF-κB) 肠道菌群 

分 类 号：R692.3+1[医药卫生—泌尿科学] R692[医药卫生—外科学] S941.42+7[医药卫生—临床医学]