小胶质细胞表达组胺受体及其在实验性变态反应性脑脊髓炎中功能调节的研究

Histamine is a potent mediator of inflammation and a regulator of innate and adaptive immune responses. Histamine has a diverse effect on many cell types due to differential expression of its receptors. However, the influence of histamine on microglia, the resident immune cells in the brain, and microglia related brain inflammation remains uninvestigated. In our study, we reported for the first time that all four types of histamine receptor proteins expressed in primary cultured microglia. Preparation of the rat primary cultured microglia. Identify the expression of mRNA and protein levels of histamin receptors on microglia. To reveal that histamine is the regulator of microglia activation and the histamine receptors expressed on microglia are the inductor to microglia activation. The effect and mechanism of histamine and H1, H2, H3 and H4 receptors on the microglia activation was examined by ELISA, DCFH-DA, MitoProbeTM JC-1 assay kit, realtime RT-PCR, westen blotting, flow cytometry analysis. The LPS brain inflammation and EAE models are prepared. The rats are intracerebroventricular injected with histamine and the agonists/antagonists of four histamine receptor subtypes. The ethology, pathology and pharmacology methods are applicated to illustrate the regulatory effect of histamine and the agonists/antagonists of histamine receptors on microglia activation in the overall level and evaluate the therapeutic effect of agonists/antagonists of four histamine receptor subtypes on LPS brain inflammation and EAE models, which suggesting that the histamine receptors may be involved in the CNS immune inflammation by regulating the microglia activation. The research results of this project will deepen the understanding of the regulation to microglial activation, improve the theory of the etiology of EAE, and also provide novel target for exploring the clinical treatment and researching & discovering ideal drug to CNS immune inflammation - related diseases.

组胺受体对参与神经免疫性炎性的多种细胞都有调节作用。然而小胶质细胞做为参与神经炎性反应的重要细胞，其是否表达组胺受体，组胺及其受体是否调节小胶质细胞活化仍知之甚少。我们的预实验结果提示小胶质细胞表达组胺受体，且组胺可以调节小胶质细胞活化。因此我们提出假说：组胺及其受体可能通过调节小胶质细胞活化参与神经系统免疫性炎症相关疾病。为了验证这一假说我们将通过大鼠原代小胶质细胞、LPS脑炎症和EAE大鼠模型，采用real time PCR、Western blot、ELISA、流式细胞仪等手段，从分子、细胞和整体水平多方面探讨组胺及其受体可通过调节小胶质细胞的活化，对LPS脑炎症和EAE大鼠模型发挥治疗作用。本项目研究成果将深化对小胶质细胞活化调节的认识，完善EAE的病因学说，为探索神经系统免疫性炎症的临床治疗途径和研发理想的治疗药物提供新靶标。

组胺受体对参与神经免疫性炎性的多种细胞都有调节作用。然而小胶质细胞做为参与神经炎性反应的重要细胞，其是否表达组胺受体，组胺及其受体是否调节小胶质细胞活化仍知之甚少。我们的实验结果提示小胶质细胞表达组胺受体，且组胺可以调节小胶质细胞活化。因此我们提出假说：组胺及其受体可能通过调节小胶质细胞活化参与神经系统免疫性炎症相关疾病。为了验证这一假说我们通过大鼠原代小胶质细胞、LPS脑炎症和EAE大鼠模型，采用real time PCR、Western blot、ELISA、流式细胞仪等手段，从分子、细胞和整体水平证明小胶质细胞表达组胺受体，并多方面探讨组胺及其受体可通过调节小胶质细胞的活化，对LPS脑炎症和EAE大鼠模型发挥治疗作用。本项目研究成果将深化对小胶质细胞活化调节的认识，完善EAE的病因学说，为探索神经系统免疫性炎症的临床治疗途径和研发理想的治疗药物提供新靶标。