脂联素通过抑制RIP1/RIP3表达对糖尿病肾病小鼠肾脏的保护作用

Protective effect of adiponectin on the kidney in diabetic nephropathy mice by inhibiting the expression of RIP1/RIP3

目的探讨脂联素对糖尿病肾病小鼠肾脏的保护作用。方法选择30只雄性健康C57BL/6小鼠作为研究对象,其中10只小鼠作为对照组,20只小鼠采用链脲佐菌素(STZ)构建糖尿病肾病(DN)模型。Apn组DN小鼠单次腹腔注射2 mg/kg Apn,每天注射1次,持续6周。STZ处理后10周检测各组小鼠体质量(BW)、左肾质量(KW)、左肾质量/体质量(KW/BW)、平均肾小球体积(MGV),检测肌酐清除率(Ccr)、24 h尿蛋白。采用酶联免疫吸附试验(ELISA)法检测小鼠肾脏组织TNF-α、IL-6的水平。采用苏木精-伊红(HE)染色观察和评定小鼠肾脏组织的病理变化。采用Western blot法检测小鼠肾脏组织中RIP1、RIP3蛋白的表达水平。结果18只小鼠成功构建DN模型,死亡2只。模型组和Apn组小鼠的BW低于对照组,KW、KW/BW以及MGV均高于对照组,且Apn组小鼠的BW比模型组高,KW、KW/BW以及MGV比模型组低,差异均有统计学意义(P<0.01)。模型组和Apn组小鼠的肾脏组织中的TNF-α、IL-6水平以及RIP1、RIP3蛋白的表达水平均明显高于对照组,且Apn组小鼠肾组织中TNF-α、IL-6水平以及RIP1、RIP3蛋白的表达水平均明显低于模型组,差异均有统计学意义(P<0.05)。结论脂联素可能通过RIP1/RIP3通路抑制细胞凋亡坏死,从而减轻糖尿病肾病肾组织的炎症反应和损伤,延缓肾组织的增生,保护肾脏组织。

Objective To explore the protective effect of adiponectin on the kidney of diabetic nephropathy mice.Methods Thirty male healthy C57BL/6 mice were selected as the research objects.Ten mice were chosen as control group,and the other 20 mice were treated with streptozotocin(STZ)to construct the diabetic nephropathy(DN)model.DN mice in Apn group were intraperitoneally injected with 2 mg/kg Apn once a day for 6 weeks.Ten weeks after STZ treatment,body weight(BW),left kidney weight(KW),left kidney weight/body weight(KW/BW),mean glomerular volume(MGV)and creatinine clearance rate(Ccr),24 h urine protein were compared.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of TNF-αand IL-6 in mouse kidney tissue.Hematoxylin-Eosin(HE)staining was used to observe the pathological changes of kidney tissue.Western blot was used to detect the expression levels of RIP1 and RIP3 proteins in kidney tissue.Results A total of 18 mice were successful to establish DN models,and two mice died.BW in model group and Apn group were significantly lower than that in control group(P<0.01),while KW,KW/BW and MGV were significantly higher(P<0.01).BW in Apn group was higher than that in model group,while KW,KW/BW and MGV were statistically lower(all P<0.05).The levels of TNF-α,IL-6 and RIP1 and RIP3 proteins in the kidney tissues in model group and Apn group were significantly higher than those in control group(P<0.05),and the levels of TNF-α,IL-6 and RIP1 and RIP3 proteins in the kidney tissue in Apn group were significantly lower than those in model group(P<0.05).Conclusion Adiponectin may inhi-bit the cell apoptosis and the necrosis through the RIP1/RIP3 pathway,thereby reducing the inflammatory response and the damage of kidney tissue in diabetic nephropathy,delaying the proliferation of the kidney tissue,and protecting the kidney tissue.