Toll-like receptor 3 (TLR3) 藉著辨識病毒產物雙股RNA (dsRNA)，引起抗病毒的免疫反應並且促使inflammatory cytokines 像是interleukin-1β (IL-1β) 和interleukin-18 (IL-18) 的產生。現在已知人工合成polyinosinic-polycytidylic acid (polyI:C) 能夠有效的刺激巨噬細胞產生interleukin-1β (IL-1β)。在本篇的研究，我們使用polyI:C刺激巨噬細胞所產生的IL-1β，去研究訊息傳遞路徑。我們觀察到PP2 (Src family kinases的抑制劑) 可以抑制 polyI:C 刺激老鼠的巨噬細胞RAW264.7所產生的IL-1β。同時，我們也觀察到 il-1β mRNA 也有顯著降低。另外一方面，AG (an iNOS inhibitor) 和ODQ (a sGC inhibitor) 在polyI:C的合併處理下，老鼠的巨噬細胞RAW264.7所產生的IL-1β 和mRNA 表現量有下降的情形，SNAP (a NO donor) 或8-br-cGMP (a cGMP analogue) 皆能增加 IL-1β的產生 和mRNA。有趣的是，基因剃除iNOS的表現確實可以使得polyI:C 刺激巨噬細胞產生IL-1β的釋放量及Src表現量降低。使用了弱化Src的src-specific siRNA細胞會使的polyI:C和SNAP所刺激的IL-1β釋放量下降，不過使用了放回ectopic Src的細胞則又能恢復IL-1β的產生。根據這些結果，我們可以得到一個結論，那就是iNOS參與polyI:C-induced IL-1β production，而且Src在這個過程中也扮演重要的角色。
TLR3 identified as recognizing of double-stranded RNA (dsRNA) and cytokines such as interleukin-1β (IL-1β) and interleukin-18 (IL-18). IL-1β is a important cytokine in macrophages by trigger the synthetic analog of double-stranded RNA (dsRNA), polyinosinic-polycytidylic acid (polyI:C). In this study, we investigated the signal transduction pathways in IL-1β secretion promoted by polyI:C . We observed that polyI:C-mediated IL-1β production was PP2 (an inhibitor for Src family kinases (SFKs)-sensitive) in murine RAW264.7 cells. Pharmacological blockade of AG (an iNOS inhibitor) and ODQ (a sGC inhibitor) decreased polyI:C-evoked IL-1β secretion. Either SNAP (a NO donor) or 8-br-cGMP (a cGMP analogue) could retrieve these defects. Interestingly, iNOS suppressed IL-1β secretion and Src induction by polyI:C in iNOS knockout mice. In particular, attenuation of Src by src-specific siRNA reduced polyI:C-IL-1β production in macrophages, and reintroduction of avain Src could rescue the defect. With these results, we concluded that iNOS was central for polyI:C-mediated IL-1β secretion and Src played a critical role in the course.
