| Background:Diabetic kidney disease(DKD)is one of the main complications after DM involves microvessels.DKD is not only the cause of increased mortality in diabetic patients.It is also the leading cause of End-stage kidney disease(ESRD).Due to the numerous and complex pathogenesis,it is still unknown,and the clinical lack of specific treatment means for DKD,therefore,once ESRD is developed,it is often more complicated and difficult to treat than other kidney diseases.Therefore,early clinical detection and identification of DKD,timely prevention and treatment are of great significance for delaying the progression of renal function and improving the quality of life and outcome of patients.DKD was previously thought to be glomerular disease,studies have shown that renal tubulointerstitial fibrosis is the main pathological change in the development of DKD to ESRD,and the epithelial-mesenchymal transition(EMT)of renal tubulointerstitial fibrosis plays an important role in the occurrence and development of renal tubulointerstitial fibrosis.With the deepening of research,attention has been paid to the role of apoptosis and autophagy of renal tubular epithelial cells in the development of DKD.High glucose and excess energy in DKD patients will inhibit autophagy,leading to cell dysfunction and apoptosis.Human umbilical cord mesenchymal stem cells(h UCMSCs)are one of mesenchymal stem cells(MSCs)with high multidirectional differentiation ability,which can secrete a variety of growth factors and cytokines.It also has certain systemic anti-inflammatory,immune suppression,damage repair,promote angiogenesis,prevent cell apoptosis.Studies have found that MSCs can improve the expression of autophagy in kidney cells and islets and improve cell function after long-term exposure to HG.Therefore,h UCMSCs and Human Kidney-2 cells,which have a wide range of sources,were used as in vitro cell lines to investigate whether MSCs can restore the function of renal cells under high glucose condition.To investigate the effect of h UCMSCs on HK2,30 mM high glucose medium was used to simulate the high glucose environment in DKD.Objective:In this study,h UCMSCs and HK2 were used as experimental cell lines to investigate the effect of 30 mM high glucose on the viability and autophagy of HK2 cells,and a co-culture system of h UCMSCs and HK2 was established to investigate the effect of h UCMSCs on the viability and autophagy of HK2 cells in a high glucose environment.Methods:HK2 cells were divided into four groups: low-glycemic group(LG group 5.5mM),high-glycemic group(HG group 30 mM),h UCMSCs co-culture group and mannitol group(5.5mM glucose +24.5mM mannitol).CCK8 was used to detect the cell viability of HK2 cultured at24 h,48h and 72 h in four different sugar concentration medium(5.5mM,10 mM,20 mM and 30 mM).The changes of HK2 autophagy markers(Beclin1 and p62)were detected by Western Blot.m RNA changes of autophagy markers(LC3,Beclin1 and p62)were detected by RT-PCR.Results:(1)CCK8 results showed that the cell vitality of HK2 in HG group(30mM)was significantly decreased compared with LG group(5.5mM)at 24 h,48h and 72h(P<0.05).After co-culture with h UCMSCs,the cell vitality of HK2 was increased compared with HG group,while that of mannitol group was not significantly affected.(2)Western Blot results showed that the expression of HK2 autophagy marker Beclin1 was decreased and p62 was increased in the HG group,which were reversed by co-culture with h UCMSCs,and there was no significant difference between the mannitol group and the LG group.(3)RT-PCR results showed that the m RNA expression of LC3 and Beclin1 of HK2 in HG group decreased,and the m RNA expression of p62 increased.After co-culture with h UCMSCs,its expression was reversed,and there was no significant difference between mannitol group and LG group.Conclusion:The viability of HK2 cells was decreased and autophagy was impaired in HG environment,while co-culture with h UCMSCs increased the viability of HK2 cells in HG environment and activated impaired autophagy. |
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