Study On The Association Mechanism Between JAK / STAT Signaling Pathway And Syndrome Differentiation

Objective:To observe the differences between blood heat syndrome/blood dryness syndrome/blood stasis syndrome of psoriasis vulgaris in JAK/STAT signaling pathway and in the expression of its related factors from the perspectives of the abnormality of Th cell differentiation, keratinocyte proliferation and abnormal dermal capillary growth.Method:Total participants of 44 were volunteers from dermatology department clinic of Dong Zhi men hospital; consist of 32 psoriasis vulgaris patients (fraction of them only participated in peripheral blood testing or only participated in skin tissue examination), and 12 healthy participants. Psoriasis patients were divided into 3 groups based on TCM syndrome differentiation techniques:Blood heat syndrome group, blood dryness syndrome group and blood stasis syndrome group. General information such as gender, age, course of the disease, inducing factors, recurrence condition, seasonal factors, PASI and degree of irritation were recorded. Flow cytometry was deployed to test the difference between Thl/Thl7 differentiation ratio in participants peripheral blood. Western bolt was deployed to test the difference of pSTAT/STAT4 and pSTAT3/STAT3 expression in the PBMC from the peripheral blood. Real time PCR was deployed to test the difference of T-bet and ROR-yt expression in participants PBMC. RT-PCR was deployed to test the difference of IL-22R1, JAK1, STAT3, C-myc and VEGF in participants skin tissue samples, immunohistocheimcal staining was used to test the location and the amount of IL-22R1, JAK1, STAT3, C-myc and VEGF in participants skin tissue samples.Result:(1) Th1/TM7 cell differentiation ratio is significantly higher in blood heat syndrome group than healthy participants (p<0.01). Thl ratio in blood heat syndrome group is higher than blood drynesss syndrome group (P<0.05), and Th17 ration in blood heat syndrome group is higher than blood dryness syndrome group and blood stasis syndrome group (p<0.05); and also the PASI and the degree of irritation of blood heat syndrome patients shows positive correlation with the Thl ratio (r=0.90,p<0.01) and (r=0.83,p<0.05). The expression of pSTAT4/STAT4 and pSTAT3/STAT3 were higher in PBMC of psoriasis patients compare to healthy people (p<0.001); blood heat syndrome group has higher expression of the factors than blood dryness and blood stasis group. The transcription factors T-bet and ROR-yt has higher expression in blood heat group than healthy people (p<0.01), and higher than blood dryness group and blood stasis group at the same time (p<0.05).(2) The results of peripheral blood testing of 24 psoriasis patients for Thl cell differentiation, pSTAT4/STAT4 expression and T-bet expression have shown positive correlation between Thl and Tbet (r=0.42,p<0.05), positive correlation between Thl and pSTAT4/STAT4 (r=0.69,p0.01) and positive correlation between pSTAT4/STAT4 and T-bet (r=0.80,p<0.01). The correlation test of Th17 cell differentiation, pSTAT4/STAT4 expression and ROR-yt expression suggest Th17 and ROR-yt show positive correlation (r=0.57,p<0.05), Th17 and pSTAT4/STAT4 show positive correlation (r=0.49,p<0.05); pSTAT4/STAT4 and ROR-yt show positive correlation (r=0.57,p0.01).(3) In the tissue of blood heat syndrome psoriasis patient, the level of gene transcription all increased and significantly higher than healthy participant (p<0.01). In the skin lesion of blood heat syndrome psoriasis patients the level of are significantly higher than healthy group and blood stasis group. In blood dryness group, protein expression of IL-22R1, JAK1, STAT3, C-myc and VEGF are higher than blood stasis group and healthy group (p0.05). In blood stasis group, the expression of STAT3/pSTAT3 is higher than normal group (p<0.05).(4) The gene transcription correlation test among 24 psoriasis patients show that IL-22R1, JAK1, STAT3, C-myc and VEGF are all positively correlated to each other (p<0.05) and the protein expression of IL-22R1,JAK1,STAT3,C-myc and VEGF are all positively correlated to each other (p<0.001).Conclusion:(1) Th cell differentiation:the activation of JAK/STAT4 and JAK/STAT3 signaling pathway can increase the expression of transcription factor T-bet and RORyt; thus mediate the increase the ratio of Thl and Th17 cell differentiation. KC and dermal capillary proliferation:ligands combine with IL-22R1 activates JAK1/STAT3 signaling pathway thus increase C-myc and VEGF expression, the result suggests that the process of Th cell differentiation, KC and dermal capillary proliferation go through JAK/STAT signaling pathway.(2) The JAK/STAT signaling pathway is most active among blood heat syndrome psoriasis patients in the pathological process of Th cell differentiation, KC and dermal capillary proliferation. JAK/STAT pathway induces inflammatory cell such as Thl/Th17 differentiation, followed by proliferation of KC cell which further lead to dermal capillary abnormal growth and deformity. The active status of JAK/STAT signaling pathway could be an important factor in the formation of psoriasis blood heat syndrome. While the JAK/STAT signaling pathway may play an important part in blood heat syndrome truning into blood dryness syndrome and blood stasis syndrome.