Effects Of Xiaoyinjiedu Granules On JAK/STAT Signal Transduction Pathway Of Jurkat T Cells

ObjectiveTo observe Xiaoyin Jiedu Granule,pharmacological serum prepared from rat as carrier acts on the pathological cell model of psoriasis(PS)which was established by Jurkat T which was induced by(phytohaemagg lutinin)PHA and(Interleukin-6)IL-6.Then,by comparing the therapeutic effects of Xiaoyin Jiedu Granule as well as western medicine and blockers of JAK/STAT pathway on JAK/STAT pathway of Jurkat T cell lines.Exploring the scientific basis of Xiaoyin Jiedu Granule in treating PS at cellular level and elucidating the immune mechanisms and targets of cooling blood and detoxicification in treating PS,providing theory basis and methods of classic tragitional Chinese medicine(TCM)prescriptions for PS under the TCM theory in clinic.Methods1.Prepare classic TCM pharmacological sercum from animal,including Xiaoyin Jiedu Granule complete ingredients group(XG),cool party group(CG),blood detoxification party group(DG),acitretin group(AG)and water control group(WG).2.PHA(10 μg/ml)activated Jurkat T lymphocytes cultured in vitro,then the CCK-8 assay was adopted to detect the proliferation of the T cells so as to determine the optimal serum concentration of Human recombinant IL-6,and the optimal inhibition concentration of pharmacological serum and blockers which stimulated T cells induced by IL-6.3.Collect Jurkat T lymphocytes induced by PHA and IL-6,which were processed by pharmacological sercum.The mRNA and proteins of the JAK1、STAT3、GP130 genes of above those cells were detected by using real-time reverse transcription polymerase chain reaction and Western blot analysis.Results1.IL-6 could promote the proliferation of Jurkat T cells,and the most obvious is 10 ng/ml;2.Compared with blank control group,concentration of 5%、10%、15%、20%all could inhibit the proliferation of Jurkat T cells significantly(P<0.001),and the most obvious is 20%;3.The blocker added could affect the survival rate of Jurkat cells in a certain extent,which had a significant statistical difference(P<0.01).50 μm/ml was the optimal inhibition concentration after analyzing cell cycle and apoptosis by flow cytometry;4.Compared with the blank cell group,the expression of JAK1,STAT3,GP130,and mRNA were increased in the cells induced by PHA+IL-6,which had a very significant difference(P<0.001);5.Adding TCM decoction、demolition,、acitretin,、distillation water pharmacological serum and pathway inhibitor into the model group(PHA+IL-6 group),the expressions of the protein and mRNA of JAK1、STAT3、GP130 were cut in these cells(P<0.05).Conclucions1.Jurkat T lymphocytes induced by PHA and IL-6 could establish the pathological cell model of PS in vitro successfully;2.IL-6 might open the JAK/STAT signal transduction pathway;3.Both the pharmacological serum of TCM and western medicine as well as blockers could inhibit the proliferation of Jurkat T cells.when the serum concentration was 20%,the inhibitory effect of the whole group was the most obvious;4.Both the pharmacological serum of TCM and western medicine as well as blockers could block the JAK/STAT pathway,and the effect of TCM was same to AG;5.To the intensity of the blocking effect on the STAT3of the pathway,CG is the same to block group,and better than XG.CG occupy an important position in the whole Xiaoyin Jiedu Granule prescription.