The Role Of Glutamate Signalling Channel In The Lymphocytes And Immunopathogenesis Of Vitiligo

Background:Studies have shown that lymphocytes express metabotropic glutamate receptors, and their activity in lymphocyte mediate a dual role in lymphocyte activation. Lymphocyte mediated mechanism to regulate immune response by glutamate receptor. Researches also confirmed that NMDA can change the lymphocyte Ca2+permeability and the level of reactive oxygen species, and this function is related with lymphocyte activation. Use NMDA and interleukin-2 commonly acting on lymphocytes, which increases NMDAR expression. NMDAR antagonists inhibit lymphocyte proliferation, and this inhibition is due to inhibits cell activity, not inhibits the cell cycle or lead to cell death. All above phenomenons confirmed that glutamate signaling pathways are related with the immune system functionnally.Vitiligo is a kind disease which because the damage in melanocytes, and cause skin and mucous membrane dispigment. It derives in the lost in function of malanocytes, by some unknown mechanism,may be associated with glutamate signaling pathways. Its etiology and pathogenesis as not yet clarified. At present the main theories are: melanocytes itself damage theory, autoimmune theory, the nerve chemical factors theory and genetic factors. Along with the development of molecular biology and immunology, researchment in autoimmune theories of vitiligo has been increasing. Studies show that patients with vitiligo whose peripheral blood nucleus cell, tumor necrosis factor and interferon release in vitiligo, may reduce inflammation response of patients skin lesions. Detection of serum interleukin-2 receptor (IL -2R), found in patients, the level of IL-2R significantly higher than normal, the level of IL-2R in patients with vitiligo related with the activity of disease. IL-2R level can be used to measure the severity of vitiligo and as an indicator of seriousness and the progression. These results demonstrate that the activation of lymphocytes may funcionnally play a important part in the pathogenesis of vitiligo.The depigmentation of vitiligo may related with the changs of glutamate signaling pathways in lymphocyte.The need of glutamate signaling pathway in further explores the role in the immunologic mechanism of vitiligo.Objective:To investigate the role of glutamate signaling pathway in the lymphocytes and immunopathogenesis of vitiligo.Method:1.Human peripheral blood lymphocytes were isolated and cultured.2.To determine MK-801 that is the non-competitive antagonist of NMDAR effect on CD25 expression and the level of CD25+ IFN-γ+ .Also and NMDA, as the NMDAR agonist and MK-801 effect on the level of ROS in lymphocytes, ,flow cytometry was perfomed.3.Real time PCR and flow cytometry methods of determination of express of NMDAR1 and NMDAR2A in nomal human and vitiligo patients lymphocytes.4.Immunohistochemical method observe the NMDAR1 and NMDAR2A expression in patients and normal lesions.Results:1.The flow cytometry analysis found that MK 801 could inhibit the pression of CD25 in nomal human lymphocytes.2.MK 801 could enhance the percentage of CD25+ IFN-γ+,in nomal human lymphocytes,which analyzed by flow cytometry.3.The effects of NMDA and MK-801 on ROS level in human lymphocytes analyzed by flow cytometry were found that NMDA can rise the level of ROS.4.The flow cytometry analysis found that people with vitiligo in lymphocytes NMDAR1 expression higher than nomal human.Conclusion:1.MK 801 as non-competitive antagonist of NMDAR can inhibit the pression of CD25,that suggests the glutamate signalling channel may effect on early active lymphocytes.2.MK 801 enhances the production of IFN-γin active lymphocytes.3.NMDA can rise the level of ROS in lymphocytes.4.The expression of NMDAR1 in peripheral blood lymphocyts of vitiligo. 5.Compared with nomal people, the expression of NMDAR1 and NMDAR2A in epidermal cells of skin of vitiligo have not differences.