基于PINK1/Parkin通路介导的线粒体自噬探讨泛酸对糖尿病肾病小鼠的影响

To investigate the effect of pantothenic acid on diabetic kidney disease mice based on mitochondrial autophagy mediated by PINK1/Parkin pathway

目的探讨泛酸是否通过PINK1/Parkin通路介导的线粒体自噬延缓糖尿病肾病(Diabatic kidney disease,DKD)的发展。方法12只雄性糖尿病肾病db/db小鼠适应性喂养2周后,随机分为模型组和泛酸组,各6只;同周龄近交式C57BL/6J小鼠为对照组。泛酸组按照130 mg·kg^(-1)·d^(-1)给予泛酸灌胃,对照组和模型组给予等体积蒸馏水,连续给药8周。实验结束后,采集各组血清、尿液进行生化检测。HE染色和PAS染色观察肾脏病理变化。RT-qPCR和Western-blot分别检测叉头框转录因子O1(FoxO1)、磷酸化叉头框转录因子O1(p-FoxO1)、同源性磷酸酶张力蛋白诱导激酶1(PINK1)、帕金蛋白(Parkin)mRNA和蛋白表达水平。结果与对照组比较,模型组空腹血糖、尿微量白蛋白肌酐比值(ACR)水平显著升高,肾小球扩张,糖原沉积;p-FoxO1蛋白表达水平显著升高,PINK1、Parkin蛋白及mRNA表达水平显著降低(P<0.01)。与模型组比较,泛酸组空腹血糖、尿ACR水平显著降低,肾小球扩张、糖原沉积减轻,p-FoxO1蛋白表达水平显著降低,PINK1、Parkin蛋白及mRNA表达水平显著升高(P<0.01)。结论泛酸有助于降低DKD db/db小鼠尿ACR水平,改善肾脏病理变化,可能是通过调控PINK1/Parkin通路,激活线粒体保护性自噬,延缓糖尿病肾病进展。

Objective To investigate whether pantothenic acid delay the development of diabetic kidney disease(DKD)through mitochondrial autophagy mediated by PINK1/Parkin pathway.MethodsAfter adaptive feeding for 2 weeks,12 male DKD db/db mice were randomly divided into model group and pantothenic acid group,with 6 mice in each group.Inbred C57BL/6J mice of the same age were used as the control group.The pantothenic acid group was given pantothenic acid by gavage at 130 mg·kg^(-1)·d^(-1),and the control group and the model group were given equal volume of distilled water for 8 weeks.After the experiment,the serum and urine of each group was collected for biochemical detection.HE staining and PAS staining was used to observe the pathological changes of kidney.The mRNA and protein expression levels of forkhead box transcription factor O1(FoxO1),phosphorylated forkhead box transcription factor O1(p-FoxO1),PTEN-induced putative kinase 1(PINK1)and Parkin were detected by RT-qPCR and Western-blot.ResultsCompared with the control group,the levels of fasting blood glucose(FBG)and urinary ACR in the model group were significantly increased,glomerular dilatation and glycogen deposition was observed.The expression level of p-FoxO1 protein was significantly increased,and the expression levels of PINK1,Parkin protein and mRNA were significantly decreased(P<0.01).Compared with the model group,the levels of FBG and urinary ACR in the pantothenic acid group were significantly decreased,glomerular dilatation and glycogen deposition was alleviated,the expression level of p-FoxO1 protein was significantly decreased,and the expression levels of PINK1,Parkin protein and mRNA were significantly increased(P<0.01).ConclusionPantothenic acid can reduce urinary ACR level and improve the pathological changes of kidney in DKD db/db mice.And it may play a role by regulating PINK1/Parkin pathway,activating mitochondrial protective autophagy,thereby delaying the progression of diabetic kidney disease.