Study On Mechamism Of Ligustilide To Intervent AtheroSclerosis Of Vulnerable Plaques By Way Of "HSP60- TLR4/MyD88-Macrophages"

Objective:By observing the expression of Myeloid differentiation primary response gene88(MyD88) and Toll-like Receptor 4(TLR4) in model of inflammatory cells and Inflammatory cytokines such as Tumor necrosis factor-a(TNF-a) and Interleukin-6(IL-6) in supernatant after intervention of ligustulide, to explore its stability molecular mechanisms of vulnerable plaques in the heat shock protein 60(HSP60) induced Macrophages (MΦ)。Method:Induce THP-1 by the Buddha waves acetate (PMA),the formation of the macrophages (MΦ) will be randomly divided into three groups as follows:the MO group, the HSP60 group and the ligustulide group.The first group using PMA to induce THP-1 cells to differentiate into MO,then continue to culture cells to extract cell protein and collect supernatant; The second group using PMA to induce THP-1 cells、to differentiate into MΦ,then continue to culture cells using HSP60 to extract cell protein and collect supernatant; The third group using PMA to induce THP-1 cells、to differentiate into MΦ, then using the ligustulide to pretreatment for 48h, at last use HSP60 to culture cells to extract cell protein and collect supernatant.Detecting the expression of related protein in cells(MyD88,TLR4) by Western Blot,detecting the concentrations of inflammatory factors in cell supernatant(TNF-a,IL-6)by ELISA.Results:1.Ligustulide can significantly inhibit the expression of TLR4、MyD88 protein in inflammatory MΦ and Inflammatory cytokines such as Tumor necrosis factor-a(TNF-a) and Interleukin-6(IL-6)in supernatant.Conclusion:Ligustulide may inhibit the activation of TLR4 and MyD88 protein by the way of "HSP60-TLR4/MyD88-MΦ", lower the concentration of IL-6 and TNF-a, which play a role of stability of vulnerable plaques.