Gorrelation Between The Signal Pathway Of T-bet/GATA3 And TCM Type In Psoriasis And Mechanism Of Chinese Medicine Therapy

Psoriasis is a chronic inflammatory autoimmune disorder that was mediated by cytokines of Th1 and based on multigenetic susceptibility. Th1 and Th2 type cytokines showed an imbalance in psoriasis. The ratio of T-bet/GATA3 is more objective than the single T-bet or GATA3 in reflecting the differentiation of Th1/Th2 lymphocytes.Therefore, regulation of the T-bet/GATA3 transcription may be a new way in treating diseases related with Thl/Th2, such as psoriasis. The aim of this study is to find correlation between T-bet/GATA3 signal pathway and TCM type in psoriasis, exploring mechanism of xiaoyintang. We compared the expression of proteins and observed EGF-induced proliferation, differentiation of HaCaT cells before treatment with after using xiaoyintang.PartⅠCorrelation of T-bet/GATA3 signal pathway and TCM type in PsoriasisObjective: To study correlation of T-bet/GATA3 and related signaling molecule and TCM type of syndromes in psoriasis.Methods: T-bet/GATA3 mRNA expression in peripheral blood mononuclear cells was measured by RT- PCR and the value of IFN-γ, IL-2, IL-4 and IL-10 by ELISA in all objects [including 45 patients with psoriasis vulgaris (20 cases of blood-heat, 13 cases of blood-dryness and 12 cases of blood-stasis) and 10 healthy people].Results: 1. The levels of IL-2, IFN-γ, T-bet and T-bet/GATA3 in serum of psoriatic patients was significantly higher than that of healthy controls, blood-heat type was significantly increased compare to that of blood-dryness type with blood-stasis type, and levels in blood-dryness type was higher than in blood-stasis type.2. The expression of GATA3 mRNA in blood-heat type was significantly lower than other two types and healthy controls; there was no difference among blood-dryness type, blood-stasis type and healthy controls.3. The levels of IL-4 and IL-10 in blood-heat type was the lowest in all groups. The level of IL-4 and IL-10 was lower in blood-dryness type and blood-stasis type than healthy controls, but there was no difference between blood-dryness type and blood-stasis type.4. The ratio of T-bet/GATA3 had a positive correlation with the levels of IFN-γand IL-2, which had a negative correlation with GATA3.Conclusion: The ratio of T-bet/GATA3 might be importamt to identify blood-heat type from nonblood-heat type in psoriatic patients. The study not only provide an objective basis for different psoriasis Syndromes of TCM but also provide new ideas for treatment of immune-related diseases with TCM.PartⅡClinical Effects and Changes of the Th1/Th2 lymphocyte balance in the peripheral blood in the Treatment of the Blood-heat Type Psoriasis with XiaoyintangObjective: To explore the mechanism of Xiaoyintang in treating psoriasis of the blood-heat type by observing the clinical effects and the impact on T-bet/GATA3 and some cytokines in serum.Methods To Observe the difference in PASI scores, scores of syndrome differentiation, the T-bet/GATA3 ratio and the levels of TNF-α, IFN-γ, IL-4 and IL-10 in serum in 20 patients with psoriasis of the blood-heat type before and after treatment. Results: There was significant difference in the T-bet/GATA3 ratio, the levels of TNF-αand IFN-γin serum before and after treatment (P<0.05), whereas the levels of IL-4 and IL-10 had no significant difference (P>0.05). PASI scores and the T-bet/GATA3 ratio, the secretion of TNF-αand IFN-γin patients with psoriasis were positively correlated (P<0.05).Conclusion: The results showed Xiaoyintang was effective in improving psoriasis, partly by decreasing the levels of TNF-α,IFN-γin serum and regulating the ratio of T-bet/GATA3.PartⅢThe effects of Xiaoyintang drug serum on EGF-induced proliferation and differentiation of HaCaT cellsObjective: To investigate the effects of xiaoyintang drug serum on proliferation and differentiation of HaCaT cells (a human keratinocyte cell line) that was induced by epidermal growth factor in vitro. Methods: Cultured HaCaT cells were treated with various concentrations (high, mean and low) of Xiaoyintang drug serum.The cell proliferation was assessed by MTT method,the changes of cell cycle and apoptosis rate was measured by flow cytometry. Results: The proliferation of HaCaT cells was inhibited by Xiaoyintang drug serum after 48h,and inhibition rate was raised with increasing treatment times and dosages.The high dosage group inhibited the proliferation of HaCaT cells by 59.47% and 73.76% at 48h and 72h after the incubation. Afer treatment with Xiaoyintang drug serum for 48h,the population of cells at Gl phase significantly increased,that at S and G2 phases decreased; the switching from Gl to G2 was inhibited, however the apoptosis was not affected. Conclusion: Xiaoyintang could inhibit the proliferation and induce the differentiation of keratinocytes in vitro.PartⅣDifferential proteomic analysis of xiaoyintang drug serum,s effect on HaCaT cellsObjective: To Identify target proteins on HaCaT cells.Methods: To Study the effect of xiaoyintang drug serum on HaCaT cells at protein level with differential proteomic methods. Results: The increasing (more than 2 times) proteins were ALK, ErbB4, EphA4, EphA2 and CSK. The decreasing proteins were ABL1, ACK1, AXL, BLK, EGFR, ERBB3, FGFR1, HGFR, IGF-1R, JAK3, LCK, MUSK , NGF, ROR1, ROR2, SCFR, TEC, TRKB, VEGFR2 and VEGFR3 .Conclusion: 5 kinds of proteins(VEGF / VEGFR, IGF-1R, EGFR, NGF, JAK3 and HGFR) associated with the pathogenesis of psoriasis were found. It provides a theoretical basis for treating psoriasis with Xiaoyintang.