Effects Of Bufalin On Autophagy Induced By Gentamicin In Rat Renal Tubular Epithelial Cells In Vitro

BackgroundAcute kidney injury(AKI)has emerged as a common symptom in intensive care unit.Some patients will develop chronic kidney injury(CKD)and eventually progress into end stage kidney disease(ESRD)and need for renal replacement therapy.Therefore,people have been looking for methods of diagnosis and treatment for AKI.As a representative agent for aminoglycoside antibiotics,Gentamicin(GM)is widely used in preventing gram-negative infection diseases.However,it is one of the common causes of acute kidney injury,and its clinical utilization is limited because of the serious nephrotoxicity.Given that,the purpose of this study is to investigate the effects of bufalin on rat renal tubular epithelial cells(Nrk-52e)autophagy and kidney injury molecule-1(KIM1)protein expression induced by GM in vitro.Understanding the role of bufalin in AKI aims to provide proper experimental foundation and theoretical basis.Methods1.NRK52e cells were cultured in vitro and were divided into 4 groups:control group,GM group(2mg/mL),Bufalin group(10-8mol/L)and GM+Bufalin group(GM 2mg/mL+Bufalin 10’8mol/L);2.NRK52e cells autophagy was observed by transmission electron microscopy;3.Effects of GM and bufalin on the expression of autophagy-related protein LC3 and P62,and KIM1 protein were examined by Western Blot.Results1.Electron microscopy showed that:Compared with control group,the number of autophagosome in GM group was significantly increased,the number of autophagosome of Bufalin group was less than that in GM group.2.Western Blot results indicated that:Compared with control group,the relative expressions of LC3-Ⅱ,P62 and KIM1 protein were significantly increased in GM group(P<0.05),but the relative expressions of LC3-Ⅱ,P62 and KIM 1 protein were less in Bufalin group than those in GM group(P<0.05).ConclusionsGentamicin could active the autophagy of NRK52e cells and upregulate the expression of KIM1 protein,while bufalin could partially inhibit GM-induced NRK52e cells autophagy and the upregulate expression of KIM1 protein,which may be a way for kidney protection by bufalin.