Effects of Triptolide on TNF-α Induced Mesangial Cell MCP-1,ICAM-1 Expression and its Mechanism

Objective:To observe the effects of triptolide on TNF-α induced secretion of MCP-1,ICAM-1 and its mechanism in glomerular mesangial cells.Methods:Cells were divided into normal control cells,TNF-α stimulated cells,triptolide (low(5 ng/ml),median(10 ng/ml) and high(15 ng/ml) concentration) incubated cells stimulated with TNF-α,PDTC intervention cells,cells incubated with both PDTC and triptolide.Supernatant MCP-1,ICAM-1,IκB,IκB α were detected using ELISA method.Cell nucleus NF-κB p65 was detected by ELISA combined with EMSA method.MCP-1 and ICAM-1 and mRNA were measured using RT-realtime-PCR.Results:In the TNF-α stimulated cells,of MCP-1,ICAM-1 Protein and mRNA,NF-κB p65 were significantly increased as compared with the control group (P0.01);In the triptolide treated cells and PDTC treated cells,MCP-1,ICAM-1 protein and mRNA,NF-κB p65 were decreased significantly(P0.01).MCP-1,ICAM-1 were decreased more significantly in the cells treated with both triptolide and PDTC as compared with the cells treated with PDTC or triptolide.NF-κB p65 positively correlated to MCP-1 and ICAM-1.IκB was decreased in the TNF-α stimulated cells.Triptolide increased IκB in a dose dependent manner.TNF-α stimulation also significantly increased IκBα phosphorylation (P0.05).But low concentration of triptolide significantly inhibited the IκBα phosphorylation (P0.05).Conclusion:Triptolide can upregulate IκB and inhibit IκBα phosphorylation to block NF-κB p65 acitivation,which then repress the secretion of MCP-1,ICAM-1 in TNF-α stimulated GMCs.