FK506治疗大鼠早期膜性肾病的实验研究

The effect of FK506 on the early experimental membranous nephropathy rats

目的观察FK506治疗大鼠膜性肾病的疗效。方法复制Wistar大鼠膜性肾病模型。实验分为3组:正常组(N组),模型组(M组),FK506治疗组(FK506组),每组12只。用阳离子化牛血清白蛋白制作大鼠膜性肾病模型,于正式免疫开始后2、4、6、8 w检测大鼠24 h尿蛋白定量;最后心脏取血,测定血清肌酐、尿素氮、白蛋白水平。处死大鼠后取肾观察肾组织病理改变;应用免疫组织化学方法检测肾组织中TGF-β1与NF-κB的表达,应用Western印迹法测定大鼠肾组织中TGF-β1蛋白表达的变化。结果M组与N组相比出现大量蛋白尿,血清白蛋白水平降低。免疫组化检测结果示M组TGF-β1和NF-κB的表达均高于N组,而FK506组的水平低于M组。Western印迹法显示M组及FK506组TGF-β1高于N组,FK506组的水平低于M组。结论FK506能降低膜性肾病大鼠尿蛋白,提高血清白蛋白,可抑制肾组织TGF-β1与NF-κB的表达,用其防治大鼠早期膜性肾病有较好的疗效。

Objective To investigate the effect of FK506 on the membranous nephropathy in rat.Methods Membranous nephropathy rats were induced by C-BSA.Wistar rats were divided into normal control（N）,membranous nephropathy（M）,and FK506 treatment（FK506） groups.Urinary protein excretion was measured at 2,4,6,8 w from the beginning of immunization.The rat blood was obtained via cardiac puncture,and the kidneys were removed.The blood samples were saved for measuring the levels of serum BUN,Scr and ALB.Kidney specimens were routinely processed for light microscopy.Immunohistochemistry method was performed to observe the expressions of TGF-β1 and NF-κB.TGF-β protein expression was measured by Western blot.Results Urinary protein excretion in group M was significantly higher than that in group N.The level of serum ALB in group M was lower than that in group N.Up-regulation of TGF-β1 and NF-κB expression were observed in immunohistochemical findings.Conclusions with that in M group,urinary protein excretion was significantly decreased and the level of serum ALB was significantly increased in FK506 group.Immunohistochemistry showed that the expressions of TGF-β1 and NF-κB were significantly lower in FK506 group than those in group M.The TGF-β1 protein expression was significantly lower in FK506 group than that in M group by Western blot.Conclusions FK506 can reduce urinary protein excretion,increase the level of ALB,improve kidney pathological changes,inhibit TGF-β1 overexpression and NF-κB activation in renal tissue of membranous nephropathy rats,and can be used for treatment of early membranous nephropathy.