低分子量肝素减少局灶节段性肾小球硬化鼠足细胞的脱落

Low Molecular Weight Heparin Decrease Podocytes Detachment in Focal Segmental Glomerulosclerosis Rats

目的观察整合素α3β1、α-肌营养不良蛋白聚糖(α-DG)在局灶节段性肾小球硬化(FSGS)大鼠肾小球的表达以及低分子量肝素(LMWH)对它们的影响。方法制备FSGS大鼠模型,随机将动物分为FSGS组、治疗组,另设对照组。治疗组给予LMWH410U.kg-1皮下注射,2次/d。8周后,检测各组24h尿蛋白定量(TP)、血清半胱氨酸蛋白酶抑制剂(CysC)、肾小球硬化指数(GSI);检测尿沉渣足细胞特异性标志蛋白podocalxin(PCX),尿PCX荧光细胞即为尿液足细胞(UPC);免疫荧光检测肾小球整合素α3β1、α-DG表达分布。分离肾小球,Western印迹法检测肾小球整合素α3β1、α-DG蛋白表达水平。光镜下观察肾组织病理改变。结果LMWH组TP、CysC、GSI以及病理改变较FSGS组明显改善。正常大鼠整合素α3β1、α-DG主要沿肾小球血管袢呈线性表达;FSGS组肾小球整合素α3β1、α-DG表达比对照组明显减弱;LMWH组整合素α3β1、α-DG表达较FSGS组明显增加。结论LMWH可以减少FSGS肾病大鼠尿蛋白,改善肾脏病理变化,保护肾功能。上调整合素α3β1、α-DG的表达,减少足细胞的脱落。

Aim To observe the effect of low molecular weight heparin （LMWH） on the expression of integnn α3β1 and α-dystroglycan（α-DG） in the glomeruli of focal segmental glomerulosclerosis（FSGS） rats. Methods SD rats were divided into three groups：normal rats, FSGS rats, FSGS rats treated with 410U·kg^-1 LMWH by subcutaneous injection twice a day for 8 weeks. 24-hour total proteinuria（TP）, serum cystatin C（Cys C）, glomerulosclerotic indexes （GSI） were measured. Podocyte-specific marker protein-podocalxin （PCX） was detected in urinary sediment and urinary PCX-positive cells were identified as UPC. The expression of integrin α3β1 and α-DG in glomeruli was determined by immunofluorescence and westernblot. In addition, the renal tissue pathology was observed by light microscopy. Results Compared to FSGS group, the levels of TP, CysC, GSI in LMWH treated group decreased significantly. Renal pathologic changes of the glomeruli in LMWH treated group were also improved. The expression of integrin α3β1 and α-DG was mainly distributed along glomerular vessels in a linear pattern. Compared to normal group, the expression of integrin α3β1 and α-DG decreased in glomeruli of FSGS rats. After g-week treatment, LMWH significantly up-regulated the expression of integrin α3β1 and α-DG compared to the FSGS rats. Conclusion LMWH can attenuate proteinuria, improves renal pathology,protects against renal function,decrease podocytes detachment and possibly by up-regulating the expression of integrin α3β1 and α-DG.