Effect Of Nuclear Factor-Kappa Bp65 On Graphene-Induced Epithelial Transdifferentiation In Human Glomerular Podocytes

Objective Recently,Hou Fanfan and his team have found that long-term exposure to smog increases the incidence of membranous nephropathy,but its intrinsic pathogenesis has not been explained,and there is no previous literature.However,previous reports indicate that podocyte injury is the last barrier of glomerular hemofiltration.The damage of podocytes is involved in almost all proteinuria-related nephropathy and plays a key role in the development of kidney disease.Among them,podocyte epithelial mesenchymal transition(EMT)plays an important role in podocyte injury.Methods The podocytes were cultured in vitro and grouped into groups:(1)podocytes were cultured in vitro and grouped with graphene concentrations of 0,10,20,40 ug,and the podocytes were stimulated at different concentrations for 24 h.(2)Real-time quantitative PCR and Western BLOT detection of EMT-related protein ZO-1、Nephrin、E-cadherin and α-SMA、Desmin mRNA and protein expression,Western BLOT detection of signaling pathway protein NF-κB p65 expression(3)After using NF-κB specific inhibitor BAY 11-7082,Real-time quantitative PCR and WesternBLOT detection of EMT-related proteins Nephrin、E-cadherin and α-SMA、Desmin mRNA and protein expression(4)Immunofluorescence staining was used to observe the changes of podocyte-specific proteins such as Nephrin and a-sma stimulated by graphene.Results Compared with the control group,the protein expression of the pathway protein NF-κBp65 was increased,the expression of EMT-related protein ZO-1、Nephrin、E-cadherin protein was decreased,the expression of ZO-1、Nephrin、E-cadherin m RNA was decreased by RT-PCR,and the α-SMA、Desmin expression of protein and mRNA was increased.Compared to 20ug/ml graphene concentration,after using NF-k Bp65 signaling pathway specific inhibitor BAY-11-7082,the expression of Nephrin、E-cadherin protein and mRNA increased,and the expression of α-SMA、Desmin protein and mRNA decreased.Conclusion Graphene promotes Epithelial-mesenchymal differentiation of glomerular podocytes via NF-κBp65 signaling pathway,and inhibition of NF-κB signaling pathway can inhibit epithelial transdifferentiation of podocytes.