To Study The Association Mechanism Of Syndrome Differentiation Of Psoriasis By Differentiation Of T Helper Cells

Psoriasis in Traditional Chinese Medicine (TCM) is known by the name of " Bai Bi", the core of its syndrome differentiation and treatment is in "blood ". The syndrome are often clinically classified into blood-heat, blood-stagnation and dryness in blood; then treated accordingly by the methods of cooling blood, promoting blood circulation and enriching blood based on skin conditions and other clinical manifestations. We have observed that TCM treatment has satisfying results. The mechanism on the onset of psoriasis is complicated and a lot is yet to be understand, this disease has the tendency to take root on young adult and its chronicle characteristic has severe adverse effect on patients’ quality of life. The recent researches have agreed that psoriasis is a genetically related autoimmune disorder which involves in the abnormality of the T cells.This article consists of two parts:literature review and laboratory research report.The literature review consist of three paragraphs, the first summarized the diagnosis and treatment of psoriasis from ancient and modern TCM school. Part two summarized TCM treatment of psoriasis and research progress of recent years regarding to T cell immunology. The third part is about the research of psoriasis of recent years regarding to Th helper cell differentiations, B cell, NK cell, miRNA aspect to narrate each immunology indicators plays its part in the onset of the disease.The laboratory standard is based on 《Guiding Principle of Clinical Research on New Drugs of Traditional Chinese Medicine in Treatment of Psoriasis.》 in 《Guidelines for New Drug Clinical Medicine》, the techniques deployed were flow cytometry, molecular biology and immunohistochemical staining. The immunology indications were measured by the clinical manifestation and peripheral blood drawn from psoriasis patients whos TCM symptom diagnosis were:blood-heat, blood-stagnation and dryness in blood.Objective:To observe the difference of specific related factors in blood of patients who were diagnosed with blood-heat, blood-stasis and dryness in blood syndrome of psoriasis vulgaris; then to find the differences in express of each syndrome---to provide an explanation in immunology mechanism and the connection between cellular immune disorder and syndrome differentiation; and further more provides a guideline in choosing treatment plan and prescription clinically.Methods:67 psoriasis vulgaris patients from dermatology department and 21 operation individuals from outpatient department of DongZhiMen hospital. Psoriasis patients enrolled in the study fit the TCM diagnosis of blood-heat, blood stagnation and dryness in blood symptoms. Flow cytometry technique used to test the ratios of T lymph cells (CD3+, CD4+ and CD8+) and Th helper cell (Th1, Th2, Treg, Th17 and Th22) in the peripheral blood taken from patients. RT-PCR used to determine the amount of RNA presence of Th cell related transcription factors (T-bet、GATA3、Foxp3、ROR γt、AHR). ABC-ELISA were used to test the level of IFN-γ、IL-4、IL-10、TNF-α、IL-17、IL-22. Immunohistochemical staining to test the amount of IFN-γRa、IL-4Ra、Foxp3、 IL-17R and IL-22R1 receptors and cell factor from abnormal skin tissue taken from all groups of patients.Result:In the peripheral blood of blood-heat syndrome patients, the ratio of Th1/Th17/Th22 cells and related cell factors (INF-γ、TNF-α、IL-22) and specific transcription factors (T-bet、ROR γt、AHR) are all higher than normal populations. (p all<0.05); at the same time, the ratio of Th2/Treg cells, interleukin (IL-4、IL-10) and specific transcription factors (GATA3 and Foxp3) are lower than normal populations (p<0.05). In blood-heat syndrome, Th1/PASI has posit ive correlation (r=0.36 p=0.02). In blood-stagnation group, the ratio of Th1/Th17 cells and related factors (INF-γ、TNF-α、IL-22) and specific transcription factors (T-bet、ROR γ t) are higher than normal population (p<0.05), whereas the ratio of Th2 cell and related cell factors IL-4 and GATA3 are lower than normal (p<0.05). The ratio of Treg/Th22; related factor IL-10 and transcription factors Foxp3 and AHR did not show significant difference from normal population (p>0.05). In dryness of blood group, the ration of Th22 and related factor IL-22 and transcription factor AHR were higher than normal (p <0.05). The ration of Th2, Treg and interleukin (IL-4, IL-10) and specific transcript ion factors (GATA3, Foxp3) are lower than normal (p<0.05). The ration of Th1/Th17 and cell factors(INF-γ、TNF-α) and specific transcription factors (T-bet, ROR γ t) shows no significant difference from normal group (p>0.05)In abnormal skin tissue test, blood-heat groups shows risen expression of INF-γRa、IL-17R and IL-22R1 (p<0.05); and lowering expression of IL-4Ra and Foxp3(p<0.05). In blood-station group, the expression of IL-17R risen (p<0.05), lower IL-4Ra expression (p<0.05); and no significant differences are shown in INF-Ra、Foxp3、IL-22R1 expression (p>0.05). In dryness in blood group, IL-22R1 expression is higher (p<0.05), IL-4Ra and Foxp3 express lower than normal group (p<0.05); INF-γ Ra、IL-17R expression show no significant differences than normal group (p>0.05)In 67 cases of psoriasis patients, cell ration correlation analysis shows Thl, Thl and Th22 cell has positive correlation (R=0.48,0.22, P=0.02,0.00), and negative correlation with Treg cell (r=-0.25, p=0.02). Th17 cell has negative correlation with Treg cell(R=-0.21, p=0.05). Thl cell ratio mainly affected by Thl7 cell, and show positive correlation with it; Thl cell shows negative correlation with Treg cell. Cell ratio correlation analysis (Thl, Th2, Treg, Th17, Th22) conducted on psoriasis patients diagnosed with blood-heat syndrome suggested Thl cell and Th17 cell display positive correlation (r=0.46, p=0.00), and show negative correlation with Treg cell (r=-0.33, p=0.04).Thl cell ratio in Psoriasis patients diagnosed with blood-heat syndrome is primarily affect by Th17 cell and show positive correlation and shows certain degree of negative correlation with Th2 cell ratio. Cell ration correlation analysis (Thl, Th2, Treg, Th17, Th22) on psoriasis patients being diagnosed with blood-stasis syndrome, the result suggested positive correlation between Th1/Th17 cell (r=0.55, p=0.04) and Th17/22 cell (r=-0.33, p=0.04). The clinical data of 67 cases of psoriasis patients (age, course of disease, degree of pruritus, PASI score, body surface area and DQLQ) is put into correlation analysis with T lymph cell sub groups and Th cells, the result shows that Thl cell has positive correlation with PASI, body surface area %(r=0.41, 0.38 P<0.05), and DQLQ (r=0.28 P<0.05). Th2 and PASI, body surface area% are in negative correlation (r=-0.25,-0.29 P<0.05). Treg and DQLQ display negative correlation (r=-0.33, P<0.05).Compare the Thl/Th2/Thl7/Treg ratio of three groups of psoriasis patients with normal subjects, the result shows Th1/Th2, Th17/Th2 ratio are significantly higher in psoriasis patients compare to normal subjects, and the ranking from high to low among 3 groups are:blood stasis> blood heat> dryness in blood. Th17/Treg ratio are significantly higher in psoriasis patients compare to normal subjects, and the ranking from high to low among 3 groups are:blood heat> blood stasis> dryness in blood.Conclusion:1 Blood-heat group shown immunology characteristic of Th1/Th17/Th22 cell differentiation advantage with Th2/Treg tune down; Thl cell can be used to predict the clinical development of blood-heat syndrome psoriasis vulgaris.2 Blood-stagnation group show immunology characteristics of Th1/Th17/cell differentiation advantage wi th Th2 tune down; Treg/Th22 cells show no significant importance in blood-stagnation psoriasis.3 Dryness in blood featuring Th22 cell differentiation advantage with Th2/Treg tune down; Th1/Th17 cells show no significant importance in dryness in blood psoriasis.4 All three types of psoriasis (blood-heat, blood-stagnation, dryness in blood) Th1/Th2 and Th17/Treg balance shift; Thl/Th2, Th17/Treg and Th17/Th2 ratio are all significantly higher than populations without psoriasis., and the severity of condition ranking from high to low among 3 groups are:blood stasis> blood heat> dryness in blood. Thl7/Treg ratio are significantly higher than normal population, and the severity of condition ranking from high to low among 3 groups are:blood heat> blood stasis> dryness in blood.5 All three types of TCM diagnosed psoriasis vulgaris displayed balance shift in Th1/Th2/Th17/Treg, and this phenomenon created the clinical progression of blood-heat> blood-stagnation> dryness in blood.6 The theory of finding treatment in "blood" shown some connection with immunologic network theory, and Th helper cell testing can help to provide feedback on different immue characteristics of psoriasis vulgaris in TCM theory.