细胞坏死性凋亡抑制剂对糖尿病小鼠肾损伤的治疗作用

Effect of necroptosis inhibitor on kidney injury in diabetic mice

目的观察细胞坏死性凋亡抑制剂Nec-1对糖尿病小鼠肾损伤的治疗作用。方法按照双盲、随机原则将48只小鼠分为对照组、糖尿病组、Nec-1对照组、糖尿病+Nec-1组,每组12只。糖尿病组、糖尿病+Nec-1组给予腹腔注射链脲佐菌素(STZ)制备糖尿病模型,Nec-1对照组、糖尿病+Nec-1组经腹腔注射Nec-11 mg/(kg·d),对照组、糖尿病组经腹腔注射等体积枸橼酸盐缓冲液。完成STZ注射后24周,收集各组24 h尿液及血液样本,将小鼠称重,处死小鼠分离肾脏并称重,计算肾质量/体质量。用全自动生化分析仪检测尿蛋白(UAE)、血肌酐(Scr)、血糖;HE染色、PAS染色、Masson染色观察肾组织病理变化;用Western blotting法检测肾组织中白细胞介素1β(IL-1β)、受体相互作用蛋白1(RIP1)、受体相互作用蛋白3(RIP3)、Ⅰ型胶原(ColⅠ)、转化生长因子β_(1)(TGF-β_(1))蛋白表达;用荧光定量PCR检测肾皮质中IL-1β、ColⅠ、TGF-β_(1)、肿瘤坏死因子α(TNF-α)mRNA表达。结果与对照组比较,糖尿病组体质量小、肾质量大、肾质量/体质量高(P均<0.05),BUN、Scr、UAE、血糖水平高(P均<0.05),肾组织病理变化重,肾组织中IL-1β、RIP1、RIP3、ColⅠ、TGF-β_(1)蛋白表达高,肾皮质中IL-1β、ColⅠ、TGF-β_(1)、TNF-αmRNA表达高;与糖尿病组比较,糖尿病+Nec-1组体质量无变化(P>0.05),以上其他指标均改善(P均<0.05);对照组与糖尿病+Nec-1组以上指标比较差异无统计学意义(P均>0.05)。结论细胞坏死性凋亡抑制剂可有效减轻糖尿病小鼠肾小管细胞炎症及肾间质纤维化,改善其肾损伤。

Objective To observe the effect of necroptosis inhibitor Nec-1 on kidney injury in diabetic mice.Methods According to the double-blind and random principle,48 mice were divided into the control group,diabetes group,Nec-1 control group,and Nec-1 group,with 12 mice in each group.Mice in the diabetes group and diabetes+Nec-1 group were given intraperitoneal injection of streptozotocin(STZ)to prepare diabetes models,mice in the Nec-1 control group and Nec-1 group were intraperitoneally injected with Nec-11 mg/(kg·d),and control group and diabetes group with an equal volume of citrate buffer solution intraperitoneally.At 24 weeks after the completion of STZ injection,24-hour urine and blood samples of each group were collected,the mice were weighed,the mice were sacrificed to separate their kidneys,and we weighed them to calculate the kidney mass/body mass.The levels of urinary protein(UAE),serum creatinine(Scr)and blood glucose were detected by automatic biochemical analyzer,and the pathological changes of renal tissues were observed by HE staining,PAS staining and Masson staining.The expression levels of interleukin-1β(IL-1β),receptor-interacting protein 1(RIP1),receptor-interacting protein 3(RIP3),collagen typeⅠ(ColⅠ)and transforming growth factor-β_(1)(TGF-β_(1))were detected by Western blotting.The mRNA expression levels of IL-1β,ColⅠ,TGF-β_(1) and tumor necrosis factor-α(TNF-α)in the renal cortex was detected by fluorescence quantitative PCR.Results Compared with the control group,the body mass was lower,but the kidney mass,kidney mass/body mass ratio,BUN,Scr,UAE and blood glucose levels were higher,and the pathological changes of renal tissue were more serious,the protein expression levels of IL-1β,RIP1,RIP3,ColⅠ,TGF-β_(1) and TNF-αin the renal tissues were higher,and the expression levels of IL-1β,ColⅠ,TGF-β_(1) and TNF-αmRNA in the renal cortex were higher in the diabetic group(all P<0.05).Compared with the diabetic group,the body weight of the diabetes+Nec-1 group did not change(P>0.05),but the other indexes were improved(all P<0.05).There were no significant differences in the above indexes between the control group and the diabetes+Nec-1 group(all P>0.05).Conclusion Necroptosis inhibitor Nec-1 can effectively reduce renal tubular cell inflammation and renal interstitial fibrosis in diabetic mice,and alleviate their renal injury.