Alpiniae oxyphyllae fructus ameliorates diabetic kidney disease through activation of PPARα-mediated autophagy
Objective:To investigate the potential protective effects of Alpiniae oxyphyllae fructus against diabetic kidney dis-ease using in vivo and in vitro models and to explore its mechanism of action.Methods:In the in vivo experiments,6 db/m mice were taken as the normal group;30 db/db mice were divided into 5 groups of 6 mice each:model,low dose Alpiniae oxyphyllae fructus,medium dose Alpiniae oxyphyllae fructus,high dose Alpiniae oxyphyllae fructus,and canagliflozin-treated.Saline was given to the normal and model groups,and the mice in the treatment groups were given the corresponding drugs respectively.After 8 weeks of treatment,serum,urine and kidney tissue samples were collected.Random blood glucose,24 h urine microalbumin,creatinine,urea nitrogen,triglyceride and total cholesterol levels were determined.Pathological changes in renal tissues were ob-served by HE staining.An in vitro model of diabetic kidney disease was constructed by intervening HK-2 cells with 60 mmol/L glucose for 72 h.After 24 h of intervention with different concentrations of Alpiniae oxyphyllae fructus-containing serum(2%,5%and 10%),the cell viability was determined by CCK8 assay and the number of autophagic lysosomes in HK-2 cells was ob-served by transmission electron microscopy.The expression levels of PPARα,LC3B Ⅱ,Beclin1,P62 mRNA and protein in mice and cells were detected by RT-qPCR and western blot.Results:In vivo experiments showed that the levels of random blood glucose,24 h urine microalbumin,creatinine,urea nitrogen,triglyceride and total cholesterol were increased in the model group compared with the normal group(P<0.000 1);the levels of the above biochemical indexes were decreased in the drug group com-pared with the model group after treatment with Alpiniae oxyphyllae fructus(P<0.05,P<0.01,P<0.001,P<0.000 1).HE staining showed that in the model group glomerular swelling increased,basement membrane thickened,renal tubules swollen and lumen narrowed.After the drug intervention,the pathological changes of the kidney were reduced to different degrees.And the ef-ficacy of both medium and high doses of Alpiniae oxyphyllae fructus was better than that of low doses.In vitro experiments showed that Alpiniae oxyphyllae fructus containing serum increased HK-2 cell viability under high glucose stress,and the best ef-fect was observed at a dose of 10%(P<0.01).Transmission electron microscopy revealed that the number of autophagic lyso-somes was decreased and lipid droplets were increased in HK-2 cells in the high-glucose group compared with the normal group;the number of autophagic lysosomes was increased and lipid droplets were decreased after Alpiniae oxyphyllae fructus intervention.RT-qPCR and western blot results in in vivo and in vitro studies were consistent,indicating that Alpiniae oxyphyllae fructus was able to increase the levels of PPARα mRNA and protein(P<0.05,P<0.01,P<0.001),and to promote the levels of Beclin1,LC3B II and decrease the expression of P62 protein(P<0.05,P<0.01,P<0.001).Conclusion:Alpiniae oxyphyllae fructus may ameliorate diabetic kidney disease by activating PPARα-mediated autophagy.
Alpiniae oxyphyllae fructusDiabetic kidney diseasePPARαAutophagyTraditional chinese medicine
万方数据
维普
