The Influence Of The Heat-clearing And Detoxifying Herbs To NF-κB Signaling Pathway In Mice Mesangial Cells Induced By TNF-α

Objective:To the Heat-clearing and Detoxifying Herbs,Honeysuckle,Weeping Forsythia, Dandelion, Herba violae, Radix Isatis,Paris rhizome, Lobelia Chinensis Lour, Smilax Glabra, Heartleaf houtluynia, Hedyotis Diffusa Willd as a research object, focusing on observation of the influence of the Heat-clearing and Detoxifying Herbs to NF-κB signaling pathway in Mice mesangial cells induced by TNF-a.discuss the effective mechanism of he Heat-clearing and Detoxifying Herbs to heat-dampness syndrome of nephropathy, which providing external basis for the use of Heat-clearing and Detoxifying Herbs in cclinicalMethods:Detecting the effect of ten Heat-clearing and Detoxifying Herbs on the cell growth by MTT colorimetric assay and select three herbs that with moderate toxic and wide role concentration range.The SV40MES13induced by TNF-a were treated with the three herbs respectively and then MTT colorimetric assay was used to detect the proliferation of SV40MES13,the amounts of the cytoplasm p-IκBα、 NF-κB p65and of nuclear NF-κB p65in SV40MES13、MCP-1and IL-6in cell supernatant were determined by ELISA.Results:(1)TNF-α (final concentration of10ng/ml) could significantly promote the proliferation of SV40MES13(P<0.05或P<0.01), the Heat-clearing and Detoxifying Herbs can inhibite the proliferation(P<0.01), the inhibition of TNF-a united the herbs is obvious than drug treatment alone, and the inhibition was in a dose-dependent manner.(2) ELISA results showed that TNF-a elevated amounts of cytoplasm p-IκBα and nuclear NF-κB p65(P<0.01),decreased amounts of cytoplasm p65markedly(P<0.01),and then facilitated nuclear translocation of p65,increased the amounts of MCP-1and IL-6in cell supernatant(P<0.01).It mained that TNF-a could induce NF-κB activation in SV40MES13and promote the level of secretion of downstream factor. The treatment groups decreased amounts of cytoplasm p-IκBα andnuclearNF-KBp65(P<0.01), increased amounts of cytoplasm p65(P<0.05或P<0.01),and then inhibited nuclear translocation of p65,decreased the amounts of MCP-1and IL-6in cell supcrnatant(P<0.01), honeysuckle had the strongest action in the three herbs with the same concentration.It mained that the Heat-clearing and Detoxifying Herbs could inhibite NF-κB activation in SV40MES13and decrease the secretion of downstream factor.Conclusion:Honeysuckle、 Weeping Forsythia ang Dandelion could inhabit the proliferation of SV40MES13induced by TNF-a and inhibit IκBα phosphorylation,reduce expresstion of NF-κB p65, and prevent nuclear translocation of NF-κB,meanwhile reduce the level of MCP-1and IL-6in cell supernatant of SV40MES13,moreover honeysuckle has the highest of inhibition in the three herbs with the same concentration. So Honeysuckle、Weeping Forsythia ang Dandelion could inhibit Rel/NF-κB/IκB signal transduction pathway.It might be the mechanism of the Heat-clearing and Detoxifying Herbs against the inflammatory injury of kidney.