Regulation Effect of Didang Decoction on HMGB1/TLR4/NF-κB Pathway
Objective:To investigate the effects and mechanisms of Dizhi Decoction and its modified formulas on inflammation in-duced by lipopolysaccharide(LPS)in human umbilical vein endothelial cells(HUVECs).Methods:HUVECs cultured in vitro were randomly divided into a control group,an LPS group,an LPS+Dizhi Decoction group,an LPS+herbal medicine group,and an LPS+animal medicine group.After inhibiting Toll-like receptor 4(TLR4),the groups were further divided into a TAK-242 group,a Dizhi Decoction TAK-242 group,a herbal medicine TAK-242 group,and an animal medicine TAK-242 group.After respective treatments,enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of high mobility group box 1(HMGB1),tumor necrosis factor-alpha(TNF-α),and interleukin-6(IL-6)in the cell supernatants.Western blot was used to determine the pro-tein expression levels of HMGB 1,TLR4,and nuclear factor kappa B p65(NF-κB p65)in HUVECs.Real-time quantitative PCR(RT-qPCR)was used to measure the gene expression levels of HMGB1,TLR4,and NF-κB p65.Immunofluorescence was used to assess the fluorescence intensity of HMGB1,TLR4,and NF-κB p65.Results:Compared with the A group,the LPS group showed in-creased levels of HMGB1,IL-6,TNF-α,TLR4,and NF-κB p65 in both the cell supernatants and HUVECs(all P＜0.01).Com-pared with the LPS group,the LPS+Dizhi Decoction group had reduced levels of HMGB1,IL-6,TNF-α,TLR4,and NF-κB p65 in both the cell supernatants and HUVECs(P＜0.05,P＜0.01).Compared with the LPS+Dizhi Decoction group,the LPS+herbal medicine group and the LPS+animal medicine group showed significantly increased levels of HMGB1,IL-6,TNF-α,TLR4,and NF-κB p65 in both the cell supernatants and HUVECs(P＜0.05,P＜0.01).The LPS+herbal medicine group had lower levels of HMGB1,IL-6,and TNF-α than the LPS+animal medicine group(P＜0.05,P＜0.01),but higher levels of TLR4 and NF-κB p65(P＜0.05).After TLR4 inhibition,compared with the LPS group,the TAK-242 group showed reduced levels of HMGB1,TLR4,and NF-κB p65(P＜0.05,P＜0.01).Compared with the TAK-242 group,the Dizhi Decoction TAK-242 group had lower levels of these indicators(P＜0.05,P＜0.01).Compared with the Dizhi Decoction TAK-242 group,the herbal medicine TAK-242 group and animal medicine TAK-242 group showed lower levels of these indicators(P＜0.05,P＜0.01).Conclusion:Dizhi Decoction and its modified formulas can protect and improve HUVECs by inhibiting cellular inflammation.The mechanism of action may be related to the inhibition of HMGB1/TLR4/NF-κB signaling pathway activation.
万方数据
维普
