Study On GCH1 Mediated ERK Pathway Regulates Microglia Activation

Objective: To study Guanosine triphosphate cyclohydrolase 1(GCH1)can participate in the activation of microglia and observe the effect of GCH1 gene expression on ERK signal transduction pathway and downstream inflammatory factors in BV2 microglia,so as to further explore the mechanism of GCH1 in neuropathic pain and provide new ideas for targeted treatment of neuropathic pain.Methods: 1.BV2 microglia with good growth condition were upregulated and downregulated by GCH1 mediated by plasmid transfection and virus interference.Microglia under different intervention conditions were divided into the following five groups: normal control group(BLANK group),empty plasmid group(NC group),GCH1 high expression group(GCH1overexpression group),empty virus group(NC siRNA group),GCH1 low expression group(siRNA GCH1 group).2.The expression of GCH1 and its effect on the activation of microglia were observed by fluorescence electron microscopy,and the difference of CD11 b expression in different groups was detected by Western Blot and RT-PCR.Western Blot was used to detect the expression of GCH1,ERK and p-ERK in BV2 microglia;RT-PCR was used to detect the expression of GCH1,ERK and p-ERK in mmra;RT-PCR and ELISA were used to detect the expression of TNF-? and IL-6.Results: 1.The high and low expression models of GCH1 in BV2 microglia could be established by plasmid transfection and virus interference.2.High expression of GCH1 can up regulate the ERK phosphorylation level of microglia,activate microglia and increase the expression level of proinflammatory factors(TNF-?,IL-6);on the contrary,low expression of GCH1 can down regulate the ERK phosphorylation level of microglia,inhibit the activation of microglia and decrease the expression level of proinflammatory factors(TNF-?,IL-6).Conclusion: The expression of GCH1 gene in BV2 microglia can be up-regulated or down regulated by transfection.The expression of GCH1 was positively correlated with the activation of microglia.GCH1 could mediate the activation of microglia by regulating ERK pathway.In the model of microglia activation mediated by GCH1,the release of IL-6 and TNF-? increased.