The most common agent of lower respiratory infections in early childhood is respiratory syncytial virus (RSV). Because there is no clear mechanism of the infection, the effective methods of therapy are not easy to develop. FIP-fve and FIP-gts are a family of immunomodulatory proteins isolated from Flammulina velutipes and Ganoderma tsugae, respectively. The molecular weight of FIPs is about 13 kDa. These family proteins stimulated the production of IFN-γ and reduced the allergic inflammation in human PBMCs. In this claim, we detect the quantity of RSV on HEp-2 cells by RT-PCR, plaque assay, Immunofluorescence assay and ELISA after FIP-fve treatment and find that FIP-fve may suppress the infection of RSV. In the claim, we detected the proteins of phospho-NF-κB and phosphor-IκBα with Western blotting and observed the translocation of NF-κB in A549 cells. These results demonstrate that FIP-fve reduce the phosphorylation of NF-κB and IκBα and the translocation of NF-κB. Furthermore, the inflammatory cytokines are decreased in HEp-2 cells with FIP-fve treatment. In vivo, oral FIP-fve and FIP-gts could alleviate on RSV-induced airway inflammation. They are developed to therapeutic agents for viral airway disease.呼吸道融合病毒(Respiratory syncytial virus,RSV)是嬰幼兒下呼吸道感染最常見的病毒,由於其感染機制目前尚未明確,因此無法發展有效的疫苗或治療方式。FIP-fve為一個從金針菇(Flammulina velutipes)純化出來的免疫調節蛋白,分子量大小為13 kDa,被歸類在真菌類免疫調節蛋白(FIPs),此類的蛋白具有調控IFN-γ增加,減少過敏發炎的功效。本研究以金針菇蛋白FIP-fve處理於感染RSV病毒之HEp-2細胞株,並分別以下列方式來偵測病毒表現量:1.聚合酶鏈鎖反應(RT-PCR)、2.空斑計數試驗(plaque assay)、3.免疫螢光染色法(Immunofluorescence)、4.酵素免疫吸附試驗(ELISA),發現在處理FIP-fve後能夠有效抑制RSV病毒的感染。另一方面,過去有研究指出,RSV會刺激細胞中IκBα的磷酸化和降解,而促使NF-κB進入細胞核中並結合到基因的轉錄因子鍵結區上,使細胞產生發炎反應。在本研究中,以西方墨點法偵測IκBα和NF-κB蛋白的磷酸化,發現處理FIP-fve後,兩者磷酸化蛋白的表現量都有被抑制的情形,進一步以共軛焦顯微鏡觀察NF-κB在細胞中的分布情況,結果顯示,FIP-fve能明顯減少由RSV所導致的NF-κB進核,而NF-κB相關的發炎因子IL-4表現量也明顯下降,綜合以上實驗結果,發現金針菇蛋白FIP-fve具有降低RSV感染,進而減少NF-κB路徑相關之發炎物質產生的療效。最後,以BALA/C小鼠進行動物模式,發現口服金針菇蛋白之小鼠,可抑制以鼻腔滴定RSV病毒後所引發的氣喘現象。AHR...呼吸道過度反應測定Methacholine(mg/ml)...乙醯甲膽鹼Penh...肺功能指標Control...控制量RSV..
