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目的:探讨糖尿病肾病发展过程中Nephrin及α-actinin-4的表达变化及其与足细胞数目的相关关系。方法:应用SD大鼠,建立1型糖尿病大鼠模型,分为糖尿病组及正常对照组,应用免疫组化及RT-PCR方法观察在糖尿病肾病发展过程中Nephrin,α-actinin-4蛋白及mRNA表达变化,以及应用WT-1来标记足细胞核,观察足细胞数目的变化。结果:8周时,糖尿病组Nephrin染色强度较正常组明显降低(P<0.05),12周时变化更为明显(P<0.01),糖尿病组α-actinin-4染色强度在两个时间点较正常组均明显增高(P<0.01),足细胞数目在两个时间点较正常组均明显减少。Nephrin表达与足细胞数目呈正相关(r=0.93,P=0.002 4),α-actinin-4表达与足细胞数目表达呈负相关(r=-0.923,P=0.008 6),而足细胞数目与尿蛋白呈负相关(r=-0.950 8,P=0.003 6)。结论:Nephrin,α-actinin-4的表达在糖尿病肾病发展过程中发生改变,此种改变与足细胞数目减少、尿蛋白的发生密切相关。
Objective: To investigate the expression of Nephrin and α-actinin-4 in diabetic nephropathy and its relationship with the number of podocytes. Methods: SD rats were used to establish a rat model of type 1 diabetes mellitus, which were divided into diabetic group and normal control group. Immunohistochemistry and RT-PCR were used to observe the expression of Nephrin, α-actinin-4 protein and mRNA in diabetic nephropathy Expression changes, and the application of WT-1 to mark the podocyte nucleus, observe the changes in the number of podocytes. Results: After 8 weeks, the intensity of Nephrin staining in diabetic group was significantly lower than that in normal group (P <0.05), and the change was more obvious at 12 weeks (P <0.01). The intensity of α-actinin-4 staining in diabetic group was significantly higher than that in normal group at two time points The normal group were significantly increased (P <0.01), podocyte number at two time points than the normal group were significantly reduced. Nephrin expression was positively correlated with the number of podocytes (r = 0.93, P = 0.002 4). The expression of α-actinin-4 was negatively correlated with the number of podocytes (r = -0.923, P = 0.008 6) Urine protein was negatively correlated (r = -0.950 8, P = 0.003 6). Conclusion: The expression of Nephrin and α-actinin-4 changes during the development of diabetic nephropathy. This change is closely related to the decrease of podocyte number and the occurrence of urinary protein.