Protective Effects of Metformin Mediated NF-κB Signaling Pathway on Cell Damage in Diabetic Nephropathy
In order to explore the protective effect of metformin mediated nuclear factor-κB(NF-κB)signaling pathway on the damage of diabetic nephropathy cells,HGPC cells were cultured in vitro and treated with 10,20,30 and 40 mmol/L D-glucose to select the optimal concentration of D-glucose for the construction of HGPC hyperglycemic inflammatory injury model.Then 20,40,80 and 160 mmol/L metformin were used for intervention,and the optimal concentration of metformin was screened.The cells were then divided into control group(Con group),high glucose group(HG group)and metformin group(Met group,30 mmol/L D-glucose+80 mmol/L metformin),inhibitor group(Y group,30 mmol/L D-glucose+1 μmol/L NF-κB pathway in-hibitor BAY 11-7082),metformin+inhibitor group(Met+Y group,30 mmol/L D-glucose+80 mmol/L metformin+1 μmol/L BAY 11-7082)and metformin+agonist groups(Met+A group,30 mmol/L D-glucose+80 mmol/L metformin+1 μmol/L NF-κB pathway agonist Prostratin),intervention was conducted for 24 h.Cell count kit 8(CCK-8)was used to detect cell viability;the expression levels of inflammatory cytokines were detected by enzyme-linked immunosorbent assay(ELISA);cell invasion and migration were determined by Transwell assay.Western blotting(WB)was used to detect the expression levels of E-cadherin,N-cadherin,vimentin,Fibronectin(FN),NF-κB p65 and p-NF-κB p65.The results show that:HGPC cells were treated with 30 mmol/L D-glucose for 24 h to construct HGPC hyperglycemic inflammation model,and 80 mmol/L was the optimal con-centration of metformin.Compared with Con group,the levels of inflammatory cytokines tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),invasion ability,migration ability,the expression levels of N-cadherin,vimentin,FN and p-NF-κB p65 protein in HG group significantly increased,while the expression level of E-cadherin protein decreased(P<0.05).Compared with HG group,the levels of inflammatory cytokines TNF-α,IL-1β,cell invasion ability,migration ability,the expression lev-els of N-cadherin,vimentin,FN and p-NF-κB p65 protein in Met group and Y group significantly decreased,and the expression level of E-cadherin protein increased(P<0.05).Compared with the Met group,the above indexes changed more significantly after the addition of BAY 11-7082(P<0.05),and the trend of Met+A group was opposite to that of Met+Y group(P<0.05).Metformin inhibits D-glucose-induced HGPC inflammatory response,invasion,migration and epithelial-mesenchymal transition process by blocking the activation of NF-κB pathway,and protects podocyte injury induced by high glucose.
diabetic nephropathymetforminnuclear factor-κB signaling pathwayD-glucose inductioninflammation of human glomerular podocytes
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