黄芪甲苷对2型糖尿病小鼠肾脏增殖细胞核抗原表达的作用

Effect of astragaloside Ⅳ on proliferation cell nuclear antigen in type 2 diabetic mice kidney

目的探讨黄芪甲苷(AS-Ⅳ)对2型糖尿病db/db小鼠的肾脏保护作用和对增生细胞核抗原(PCNA)的影响。方法雄性db/db小鼠,随机分为db/db组和db/db+AS-Ⅳ组,另以野生型(WT)小鼠作为对照组。AS-Ⅳ予1 g·kg-1饲料喂食。第12周,检测各组小鼠体质量、血糖、肾质量、肌酐清除率和24 h尿白蛋白定量。PAS染色和电镜检测各组小鼠肾脏病理。免疫组织化学和Western blot检测肾组织PCNA的蛋白表达水平。结果第12周时,db/db组小鼠体质量、血糖、肾质量、肌酐清除率明显高于WT组,但db/db+AS-Ⅳ组与db/db组之间差异无统计学意义。db/db组小鼠24 h尿白蛋白水平明显高于WT组,db/db+AS-Ⅳ组小鼠24 h尿白蛋白水平低于db/db组,差异有统计学意义(P<0.05)。db/db组小鼠肾小球面积,肾小管面积,肾小球基底膜厚度和肾小管基底膜厚度均显著高于WT组,db/db+AS-Ⅳ组上述指标明显低于db/db组(P<0.05)。免疫组织化学和Western blot结果显示db/db组小鼠肾组织PCNA的蛋白表达水平显著高于WT组,AS-Ⅳ可以明显降低PCNA的表达水平,差异有统计学意义(P<0.05)。结论 AS-Ⅳ可能通过减少2型糖尿病小鼠肾脏内增生细胞核抗原的蛋白表达来延缓糖尿病肾病进程。

ObjectiveTo investigate the effect of ASⅣon kidney injury in the kidney of type2db/db mice,and its effect on PCNA protein.MethodsTwelve male db/db mice were randomly assigned into two groups:one group of db/db mice fed with normal rodent chow while the other group of db/db mice fed with normal rodent chow and ASⅣ(1g·kg-1diet).The wild type mice served as control group.After12week treatment,mice body weight(BW),blood glucose(BG),kidney weight(KW),creatinine clearance(cCr),albumin excretion rate(AER)were measured.Renal pathological changes were detected by PAS staining and electron microscopy.The renal cortical protein level of PCNA was detected by western blot and immunohistochemistry.ResultsCompared with wild type mice group,the levels of BW,BG,KW and cCr were higher in db/db group.There was no significant difference in these parameters between db/db group and db/db+ASⅣgroup.Compared with wild type mice group,the excretion rate of albuminuria was higher in db/db group;the excretion rate of albuminuria was lower in db/db+ASⅣgroup.The differences were statistically significant(P<0.05).The glomerular turf area,tubular area,thickness of glomerular basement membrane and tubular basement membrane in db/db group were higher than wild type mice group;these indexes in db/db+AS IV group were lower than db/db group.The differences were statistically significant(P<0.05).Immunohistochemistry and western blot results showed that renal PCNA expression levels in db/db group were higher than that in WT group,and AS IV could significantly decrease the level of renal PCNA expression.The differences were statistically significant(P<0.05).ConclusionsThese findings suggested that AS IV can prevent the progression of diabetic nephropathy by down regulating PCNA expression level in diabetic kidney.