高脂诱导的肾脏损害-第三军医大学学报.docVIP

高脂饮食诱导CD36/SR-A/ApoE三敲除小鼠肾脏损害及其机制的初步研究 徐珍娥1，闫凤2，阮雄中2，3，陈压西2，李秋1（400014 重庆，重庆医科大学附属儿童医院肾内科1，400016 重庆，重庆医科大学感染性疾病分子生物学教育部重点实验室脂质中心2，英国UCL大学皇家自由医学院肾脏病科3） [摘要] 目的 脂质诱导肾脏损伤的机制尚不清楚，本课题的目的在于探讨低密度脂蛋白受体相关蛋白（1ow density lipoprotein receptor-related protein ,LRP1）、清道夫受体CD36、SR-A在脂质诱导肾脏损伤中的作用。方法 采用的雄性ApoE/CD36/SR-A三敲除小鼠，将小鼠随机分为普通饮食组和高脂饮食组。检测血清淀粉样蛋白（serum amyloid A protein,SAA）,IL-6、脂质水平及尿蛋白。油红“O”染色及酶法检测肾组织内脂质沉积情况。HE、过碘酸-希夫（periodic acid Schiff reaction,PAS）、马松（MASSON）方法观察肾脏形态学改变，定量PCR技术，western-blot与免疫组化技术检测肾组织相关因子水平。结果 高脂饮食组脂质水平[TC(51.96±5.60)、LDL(16.79±4.80)mmol/L]显著高于普通饮食组[TC(18.11±3.07)、LDL(3.27±1.79) mmol/L](P0.05)。肾脏脂质的沉积明显升高，高脂饮食组尿蛋白[(2.90±0.50)mg/ml]显著高于普通饮食组[(1.85±0.18)mg/ml] (P0.05)，肾脏病理改变显著，进一步检测发现高脂饮食组上调了LRP1（增高2.23倍）、肾脏I型胶原蛋白、转化生长因子（TGF-β）、纤维连接蛋白（Fibronectin）mRNA与蛋白的表达。结论 LRP1可能参与肾脏泡沫细胞的形成及肾脏的损伤，SRA,CD36不能解释泡沫细胞形成的全部机制。 [关键词] 脂质；肾小球硬化；ApoE/CD36/SR-A KO小鼠；LRP1 [中图分类号]R3 [文献标志码] A Mechanism of High Fat Diet Induced Renal Injury of CD36/SR-A/ApoE KO Mice XU Zhen’e1, YAN Fen2, RUAN Xiongzhong2，3, CHEN Ya-xi 2, LI Qiu 1（1：Nephrology department of Children’s Hospital of Chongqing Medical University, Chongqing, 400014, China,；2：Centre for Lipid Research, Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing；400016, China,） [Abstract] Objectives To investigate whether lipid mediated glomerular injury by non–scavenger receptor–mediated pathways in vivo. Methods Male ApoE/CD36/SR-A knock-out mice were randomly assigned to feed a chow diet or western diet. Animals were sacrificed after 14 weeks; terminal blood samples were taken for plasma serum cholesterol, triglycerides, LDL-cholesterol, HDL cholesterol, amyloid A (SAA) and interleukin-6 (IL-6) assays. Renal sections were used for histological and immunohistochemistry assessments. The lipid accumulation in kidney was evaluated by Oil Red O staining. The mRNA and protein expression of LRP1, TGF-β, collagen type I, Fibronectin, were analy