Effects Of Exogenous Glutamate On The Secretion Of TNF-? And IL-1? In Keratinocytes

Background: Vitiligo is the main clinical features of depigmentation,which is an intractable skin disease with high incidence,seriously affecting the patients physical and mental health.At present,most scholars believe that vitiligo is caused by the interactions between autoimmune and oxidative stress in multi-gene genetic background.The changes and interactions of cytokines in peripheral blood and skin lesions promote the progress of immune response and inflammatory response,and then induce or aggravate the occurrence of disease.Previous studies have found that high concentrations of tumor necrosis factor-? and interleukin-1? attacked to melanocytes,resulting in melanin production reduced,pigment loss.Glutamate is an important excitatory amino acid neurotransmitters in central nervous system.The researcher also found a functional glutamate signaling pathway in non-peripheral tissue in recent years.Our team's previous study found that there was functional glutamate signaling pathway in the skin.Previous study found that the expression levels of TNF-? and IL-1? were significantly higher in the vitiligo lesions skin.There were reports that these cytokines are produced mainly by keratinocytes.Therefore,the study devoted to exploring the influence of glutamate on the secretion of TNF-? and IL-1? in keratinocytes,which could provide a theoretical basis for the depth study of the pathogenesis of vitiligo.Objective:To study the effect of exogenous glutamate on the secretion of TNF-? and IL-1? in keratinocytes.Methods: 1.The expression of cysteine / glutamate transporter(System Xc-)in Ha Ca T cells was verified by real-time quantitative polymerase chain reaction.2.The content of glutamate was detected by Glutamate Assay Kit(SIGMA)in the supernatant of Ha Ca T cells.3.The expression of TNF-? and IL-1? in the supernatant of the cell culture was detected by enzyme-linked immunosorbent assay(ELISA)in Ha Ca T stimulated with different concentrations of glutamate.Results: 1.The result of real-time quantitative polymerase chain reaction methods showed that Ha Ca T expressed cysteine / glutamate transporter.2.When the content of exogenous glutamate was in the range of 0.060-0.095 m M,the secretion of TNF-? in Ha Ca T was gradually increased,but the result hadno statistically difference(P>0.05).When the content of exogenous glutamate was in the range of0.095-0.783 m M,the secretion of TNF-? in Ha Ca T was gradually decreased,which had a statistically significant difference(P <0.05).3.When the content of exogenous glutamate was in the range of 0.060-0.095 m M,the secretion of IL-1? in Ha Ca T gradually increased,which had a statistical difference(P <0.05).When glutamate was in the range of 0.095-0.783 m M,the secretion of IL-1? in Ha Ca T gradually decreased,which had a statistically significant difference(P <0.05).Conclusion: 1.Ha Ca T transfer glutamate from intracellular to extracellular via cystine / glutamate transporter.2.High concentration of exogenous glutamate inhibits the secretion of TNF-? and IL-1? in Ha Ca T,but low concentrations of exogenous glutamate promotes.