The Effect Of Retinoic Acid On The Proliferation Of Periphral T-lymphocytes In Psoriasis And Correlation Study Of JAK3 And STAT5

About 2,500 kinds of retinoid acid (RA) derivatives have been found currently,and RA at least used in the treatment of 150 diseases,especially in refractory keratosis dermatitis, such as psoriasis, a new way for physicians. Moreover, their understanding and treatment, and indications are continually expanded. The treatment of RA is called the"second revolution"in the history of dermatology therapy.The basic structure of RA is composed of three parts,including cyclohexene, side-chain and polar group ,it can be divided into three generations, that is non-aromatic, single aromatic and more aromatic. Natural RA is of the first generation, including all-trans retinoic acid (atRA), 9 - cis-retinoic acid and isotretinoin. The second generation of RA is synthetic, such as etretinate and acitretin. The notable feature of the third generation is the introduction of a number of aromatic , it is selective receptor, the targeted drugs, at the same time reduce the adverse reaction, for example,Adapalene and Tazarotene. Arotinoid trometamol is more aromatic structure,belong to a third generation of retinoid.Psoriasis is characterized by pathological changes in the proliferation and differentiation of epidermal cells, accompanied by significantly inflammatory infiltrateion. With the depth of research about the underlying pathogenesis of this diseasese, psoriasis is thought as an unusual immune disease with genetic background , abnormal activated T lymphocytes and the secretion of a variety of cytokines is the central link in the pathogenesis.Clinically, retinoid make good efficacy in the treatment of psoriasis, but the mechanisms are not yet clear. Studies suggest that it is pocess of anti-hyperkeratosis and anti-tumor effect. In recent years, more and more studies concerntrate on the role of immune regulation. Retinoid can affect many aspects of immune system, such as proliferation and differentiation of immune cells, immune activity signal transduction, cytokine production, thus affecting the cell immune function and immunoglobulin production, some aspects show bidirectional regulation, but the data about " retinoic acid is of immunity regulation", are very limited, and no reports about retinoid on immune system in normal and psoriasis.In fact, with the depth of research, this area will be becoming a new focus. The first generation of all-trans retinoic acid (atRA) and 9– cis retinoic acid, which can impact the proliferation of normal human peripheral blood T lymphocytes. The results confirmed that :the role of RA in the health of human peripheral blood T lymphocytes is a complex process, the mechanisms are diversity .Janus (Janus kinase) -/STAT (signal transducer and activator of transcription) is the cellular signaling pathways in the dominant role of cytokine signal transduction,which has a very closed relationship with the proliferation and differentiation of above,which is also an important way of lymphocyte activation.Cytokines play an important role in the cell development,differentiation and immune regulation. JAK/STAT ways and therefore the relationship between the immune system are gradually becoming to a new hot. The regulation of specific signal transduction process and the detailed research will help our in-depth understanding of the interaction of cells and the mechanisms of normal physiological processes and certain diseases. It may provide a theoretical basis for develop the pharmacological effect of drugs.Choi used retinoic acid-treated rat brain astrocytes, found all-trans and 9– cis retinoic acid inhibit JAK/STAT pathway by inhibiting the activation of IFN-γ-induced inhibition of the inflammatory response. Janus is a protein tyrosine kinase (PTK), four family members found to date : JAK1 and JAK2. JAK3, and TYK2. JAK3 in which activation of T cells, B cells and mononuclear cells with a wide range of expression. The effect can be activated by IL-2, IL-4, IL-7, in certain circumstances, Janus may also be activated by epidermal growth factor receptor (EGFR),platelet-derived growth factor receptor (PDGFR). GU found JAK3 was high expression in the peripheral blood lymphocytes with rheumatoid arthritis and arthropathic psoriasis.It is found recently that JAK pathway activation, especially STAT activation, has a major impact on the cell proliferation and transformation. The current study shows that, under normal circumstances, STAT5 maintain the normal function of cells. Regulation plays an important role in cell proliferation and differentiation. When the inappropriate activation of STAT5, caused by the downstream target genes such as over-expression of cyclin D1, c-myc and bcl-xl ,which induced in cancer and blocked in cell apoptosis.Expression of STAT5 may be associated with the development of various types of cancer, which may play an important role in immune tolerance, STAT5 deficient mice showed mild decrease in lymphocytes, T cell proliferation defect.The emergence of a new generation of receptor-selective retinoic acid significantly reduced the side effects of vitamin A derivatives,which encourage people reform retinoids structure. It is arotinoid trometamol that combined retinoic acid receptor selective RAR agonists-- aromatic retinoid and trometamol group,which synthesize a third generation of retinoid, which increase the molecular weight and soluble.The aim of this study - compare arotinoid trometamol and acitretin with dose effect ,according to inhibition of proliferation of T lymphocyte with psoriasis, the relevance of JAK3/STAT5 transcription and protein level, trying to clarify the mechanism of drug treatment and therefore provide a new targeted drugs for psoriasis.PartⅠThe dose effect of arotinoid trometamol and acitretin on the proliferation of T lymphocytes in psoriasisObjective: The effect of the third-generation tretinoin (arotinoid trometamol) and the second generation of tretinoin (acitretin) on the proliferation of T lymphocytes in patients with psoriasis in a dose-responsive manner.Method: Collectting peripheral T lymphocyte, treating with different concentrations of drug. Trypan blue staining cells, modified MTT assay detecting lymphocyte proliferation.Results: Peripheral T lymphocytes from patients were dealed with 0.1μM of the two drug, the percentage of living cells was low(not the choice of concentration). Within 10-9-10-6mol/L concentration range,the two drugs could inhibit the proliferation of human peripheral blood T lymphocytes.Compared with the control group, there were a significant meanings, and also significantly statistics between the two drugs.Conclusion: Arotinoid trometamol and acitretin could inhibit the proliferation of T lymphocytes, the effect were dose-dependent, the inhibition of Arotinoid trometamol was more obvious than acitretin.PartⅡThe effct of arotinoid trometamol and acitretin on the expression of JAK3 geneObjective: Observating the effct of arotinoid trometamol and acitretin on the expression of JAK3mRNA in T lymphocytes with psoriasis.Method: Cell accordance with 5×105 inoculated in the culture adding 10-9～10-6mmol/L Arotinoid trometamol and acitretin ,control holes without medicine, 37°C for 48 h, then extracted RNA, GAPDH/β- actin as reference, Real-time PCR method was used to detect JAK3mRNA expression.Results: Peripheral T lymphocytes were dealed with each concentration of arotinoid trometamol and acitretin for 48 hours, compared to their baseline,the effect of JAK3 transcriptional level was of significantly statistics. Conclusion: Two drugs could reduce the expression of JAK3mRNA in peripheral T lymphocytes. Arotinoid trometamol downward effect can be more pronounced.PartⅢThe effct of arotinoid trometamol and acitretin on the expression of JAK3/STAT5 proteinObjective: Observating the effct of arotinoid trometamol and acitretin on the expression of JAK3/STAT5 proteins in peripheral T lymphocytes.Method: After drug treatment, the above cell, dubbed as 5×106 per/mL cell suspension. RIPA lysate proteins and JAK3, STAT5 proteins expression were detected by Western blot.Results: T lymphocyte were dealed with two drugs for 48 hours, compared with baseline,the level of JAK3 protein expression were significantly downward; the downstream of STAT5 protein expression was of no significantly statistics.Conclusion: Arotinoid trometamol and acitretin could significantly reduce the expression of JAK3 protein, STAT5 downstream protein expression were no change.