保护素D1通过抑制肾脏炎症及足突细胞上皮-间充质转化而抑制早期糖尿病肾病小鼠肾纤维化  

Protectin D1 inhibits renal fibrosis in early-stage diabetic nephropathy mice by inhibiting renal inflammation and podocyte epithelial-mesenchymal transition

作　　者：王全胜[1] 尹红霞 张阿丽[3] 陈建武[1] 卢芙蓉[1] 薛卡明[1] 谢纪文 刘建国[1] 李仁康[1] 邓安国[4] 

机构地区：[1]华中科技大学同济医学院附属协和医院中西医结合科,湖北武汉430022 [2]荆州市妇幼保健院,湖北荆州434000 [3]武汉大学中南医院放化疗科,湖北武汉430071 [4]华中科技大学同济医学院附属协和医院肾内科,湖北武汉430022

出　　处：《中国病理生理杂志》2013年第12期2144-2151,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30600810)

摘　　要：目的:保护素D1(PD1)是一个潜在的抗炎症脂蛋白分子,本实验探讨其治疗早期糖尿病肾病(DN)肾纤维化的作用及机制。方法:用链脲佐菌素125 mg/kg 2次腹腔注射C57BL/6J雌性小鼠,建立早期DN小鼠模型。糖尿病模型成功后,用PD1(0.08 mg·kg-1·d-1)腹腔注射治疗,设正常鼠及DN鼠为对照。治疗8周后检测各组小鼠24 h尿蛋白及尿白蛋白定量、体重、肾重、肾重/体重比、血清及尿肌酐和肌酐清除率;用PAS染色法检测肾小球系膜区基质/肾小球面积比,用免疫荧光染色法检测肾皮质中巨噬细胞的数量,用Western blotting检测肾小球纤连蛋白(FN)和α-平滑肌肌动蛋白(α-SMA)的表达,以及肾小球足突细胞特异性上皮标志蛋白zonula occludens-1(ZO-1)和P-cadherin的表达;同时,体外用高糖刺激小鼠巨噬细胞株RAW264.7,检测PD1对其分泌肿瘤坏死因子α(TNF-α)和白细胞介素1β(IL-1β)的抑制作用。体外用转化生长因子β1(TGF-β1)刺激小鼠足突细胞株,用Western blotting检测PD1对其诱导足突细胞上皮-间充质转化(EMT)中上皮细胞标志蛋白P-cadherin和ZO-1减少的恢复作用,及间充质细胞标志蛋白成纤维细胞特异性蛋白1(FSP1)和α-SMA过表达的抑制作用。结果:PD1能减少DN小鼠肾小球系膜基质的积聚、24 h尿蛋白及尿白蛋白定量、体重、肾重和肾重/体重比,抑制异常增高的肌酐清除率。PD1能减少DN小鼠肾皮质中巨噬细胞的数量,抑制DN小鼠肾小球FN和α-SMA的表达,恢复足突细胞特异性上皮标志蛋白ZO-1和P-cadherin的表达。PD1能抑制高糖诱导RAW264.7分泌TNF-α和IL-1β,能抑制TGF-β1诱导足突细胞FSP1和α-SMA表达的增加以及ZO-1和P-cadherin表达的减少。结论:PD1能减轻早期DN小鼠肾纤维化,其部分机制可能通过抑制肾脏的炎症及足突细胞EMT。AIM： To study the effect of protectin D1 （PD1） as a potent anti-inflammatory lipid mediator on diabetic nephropathy （DN）. METHODS ： PD1 （0.08 mg · kg-1 . d-1 ） was intraperitoneally injected into the mice for 8 weeks after diabetes was induced by injection of streptozotocin. The 24-h urinary protein and albumin levels, body weight, renal weight, kidney-to-body weight ratio, ereatinine clearance, glomerular mesangial matrix accumulation, renal cortical macrophage accumulation, and glomerular expression of fibronectin （ FN）, cL-smooth muscle actin （α-SMA）, zonula occludens-1 （ZO-1） and P-cadherin were detected. Thfe effect of PD1 on inhibiting epithelial-mesenchymal transition （EMT） in the podocytes induced by transforming, growth factor 1~ （ TGF-13~ ）, and the effect of PD1 on inhibiting the in- flammatory effect of macrophages induced by high glucose were determined. RESULTS ： PD1 markedly suppressed diabe- tes-induced elevation of 24-h urinary protein and albumin levels, body weight, renal weight, kidney-to-body weight ratio, creatinine clearance, glomerular mesangial matrix accumulation, and glomerular expression of FN and ct-SMA. PD1 also suppressed diabetes-induced increase in the number of renal cortical macrophages in the mice with DN. Analysis by West- ern blotting and immunohistochemistry revealed that PD1 suppressed diabetes-induced elevation of mesenchymal/fibrotic markers FN and ct-SMA, and increased podocyte-related epithelial markers ZO-1 and P-cadherin in the glomeruli of the mice with DN. PD1 repressed high glucose-induced generation of tumor necrosis factor ct and interleukin 1~ by macropha- ges, and inhibited TGF-13l-induced increases in fibroblast-specific protein 1 and ct-SMA, and inhibited TGF-13t-induced decreases in epithelial markers P-cadherin and ZO-1 in podocytes in vitro. CONCLUSION： PD1 inhibits renal fibrosis in the early stage of DN, and its mechanisms may be related to its anti-inflammatory and anti-EMT effects on podocytes.

关 键 词：保护素D1 糖尿病肾病 纤维化 炎症 足突细胞 上皮-间充质转化 

分 类 号：R363[医药卫生—病理学]