| Inflammatory skin disease such as psoriasis is associated with immune disorder, all kinds of immunocytes and inflammatory mediators play an important role in the pathogenesis. The complex network of cytokines and mutual manipulation hint that single target drugs are unsuitable for therapy. So the multiple targets drugs increasingly become the focal point in therapy of psoriasis.TLR-4 is a toll-like receptor, it detects lipopolysaccharide on Gram-negative bacteria and is thus important in the activation of the innate immune system, and some research reveal that TLR-4 is associated with psoriasis immune disorder. The TLR-4 and the innate immune response mediated by it may be very important to the pathogenesis of psoriasis. Dendritic cells and mononuclear-macrophage are key sentinels of the immune system, bridging the gap between innate and adaptive immunity, they are key constituents of the psoriatic inflammatory process, plasmacytoid dendritic cells as potential drug targets of psoriasis has already been more and more study. THP-1 cells derived from the peripheral blood of a 1 year old human male with acute monocytic leukemia and can be differentiated into macrophage-like cells.Curcumin,13-hexyl-berberine hydrochloride(HB-13),a derivative of berberine and 13-hexyl-palmatine hydrochloride (HP-13),a derivative of palmatine has significant anti-inflammatory, anti-oxidant, anti-biosis, and anti-cancer properties. So these drugs may play a functional and potentially therapeutic role in inflammatory skin disease such as psoriasis. Our laboratory have researched the effects of curcumin,HB-13 and HP-13 on INF-y signal transduction in HaCaT cells. The aim of this study was to investigate the effects of those compounds on regulation of inflammatory cytokines in TLR-4 signal transduction in THP-1 cells.The regulation effects of curcumin,HB-13 and HP-13 probably contributes to the treatment of inflammatory skin disorders, such as psoriasis.Chapter I Effects of Topical application and injection of compounds on TPA-induced edema of mouse ears.Objective To investigate the effects of curcumin,HP-13 and HB-13 on TPA-induced edema of mouse ears. Methods The anti-inflammatory action of topical application and injection of curcumin, HB-13, HP-13 was observed through external ear inflammation of mice induced by TPA. Results Injection of curcumin(100mg/kg) or HP-13(20mg/kg) or HB-13(10mg/kg) has no acute toxic reaction.Topical application curcumin (0.3mg), HB-13 (0.38mg), HP-13(0.36mg)or injection of curcumin(50,100mg/kg) or HP-13(10,20mg/kg) or HB-13(5,10mg/kg) obviously inhibited TPA-induced epidermal inflammation (ear edema) in mice. The inhibition rate is above 50%(P<0.01). Conclusion Topical application or injection of a certain concentration curcumin, HB-13 or HP-13 obviously inhibited TPA-induced epidermal inflammation (ear edema) in mice. The curcumin, HB-13 and HP-13 exhibited marked anti-inflammatory activity in the animal experiment.Chapter II Effects of compounds on the expression of TRAF6, IRAK1, NF-κB, TNF-a and IL-1βmRNA in THP-1 induced by LPS.Objective To investigate the effects of curcumin,HB-13 and HP-13 on the expression of TRAF6, IRAK1, NF-κB, TNF-a and IL-1βmRNA in THP-1 induced by LPS. Methods The expression of TRAF6, IRAKI, NF-kB, TNF-a and IL-1βmRNA was examined by RT-PCR. Results The expression ability of TRAF6, IRAK1, NF-κB, TNF-a and IL-1βmRNA in THP-1 has increased significantly after THP-1 induced by LPS (1μg/mL) for 4h. Curcumin(50μg/mL),HB-13(0.39,0.78μg/mL) or HP-13(1.56,3.12μg/mL) can reduce markedly mRNA expression level of TRAF6, IRAK1, NF-κB, TNF-a and IL-1βin THP-1 induced by LPS. The inhibition rates of curcumin, HB-13, HP-13 to goal genes are all above 50% Conclusion The curcumin, HB-13, HP-13 can inhibit TLR-4 signal transduction by inhibiting the expression of TRAF6, IRAK1, NF-κB, TNF-a and IL-1βmRNA in THP-1 cells.Chapter III Effects of compounds on the secretion of downstream signal transduction proteins of TLR-4 induced by LPS in THP-1 cellsObjective To investigate the Effects of curcumin HB-13 and HP-13 on the secretion of TNF-a,IL-8,IL-1βinduced by LPS in THP-1 cells. Methods The secretion of TNF-a,IL-8,IL-1βwas detected by ELISA. Results THP-1 cells were found to secrete TNF-a,IL-8,IL-1βconstitutively. The secretion was induced by LPS, stimulation with 1μg/mL LPS for 24 hours dramatically enhanced the expression of TNF-a,IL-8,IL-1β. Pretreatment with curcumin, HB-13 or HP-13 for 12 hours dose-dependently inhibited TNF-a,IL-8,IL-1βsecretion induced by LPS and the IC50 values were:TNF-a: curcumin:30.4671μg/mL, HB-13:0.2924μg/mL, HP-13:2.0182μg/mL; IL-8:curcumin: 19.0561μg/mL, HB-13:0.5132μg/mL, HP-13:1.7766μg/mL:IL-1β:curcumin: 32.9003μg/mL, HB-13:0.4547μg/mL, HP-13:2.3403ug/mL respectively. Conclusion The curcumin, HB-13 and HP-13 can inhibit the expression of inflammatory cytokines(TNF-α,IL-8,IL-1β) induced by LPS in THP-1 cells.
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