【摘要】：為了探究SHP1在豬繁殖與呼吸綜合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)誘導(dǎo)的抗病毒免疫中的作用,本研究采用200個TCID50的PRRSV感染豬肺巨噬細胞(porcine alveolar macrophage,PAM),分別培養(yǎng)12,24,36,48h后收集細胞及上清,分別設(shè)正常細胞對照組,聚肌胞(polyinosinic-polycytidylic acid,polyI:C)刺激對照組,脂多糖(lipopolysaccharide,LPS)刺激對照組。采用ELISA方法檢測PRRSV感染的PAM培養(yǎng)上清中SHP1的蛋白表達情況。用實驗室已經(jīng)建立的實時熒光定量PCR方法對PRRSV感染組和各個對照組PAM細胞中IRF3、IRF7、TRAF6、NF-κB、SHP1和IFN-β的mRNA轉(zhuǎn)錄水平變化進行定量分析。結(jié)果顯示:激活的SHP1對PRRSV感染后PAM中IRF7、NF-kB和IFN-β的轉(zhuǎn)錄有促進作用,對IRF3和TRAF6的轉(zhuǎn)錄有抑制作用。LPS及Poly(I:C)刺激PRRSV感染的PAM細胞均促進了IRF3、IRF7、TRAF6、NF-κB、SHP1和IFN-β的轉(zhuǎn)錄。結(jié)果表明,激活的SHP1會通過TLR介導(dǎo)的信號通路或未知通路來調(diào)節(jié)PRRSV誘導(dǎo)的抗病毒免疫反應(yīng)水平。LPS和Poly(I:C)刺激PRRSV感染的PAM細胞能夠增強抗病毒細胞因子的轉(zhuǎn)錄水平。
[Abstract]:To investigate the role of SHP1 in porcine reproductive and respiratory syndrome virus (porcine reproductive and respiratory syndrome virus,PRRSV) -induced antiviral immunity, 200 TCID50 PRRSV were used to infect porcine pulmonary macrophages (porcine alveolar macrophage,PAM). The cells and supernatants were collected after 48 hours of culture, and were divided into normal cell control group, polymyocyte-stimulated control group (polyinosinic-polycytidylic acid,polyI:C) and lipopolysaccharide (lipopolysaccharide,LPS) stimulated control group. ELISA method was used to detect the expression of SHP1 protein in the supernatant of PAM infected with PRRSV. The changes of mRNA transcription levels of IRF3,IRF7,TRAF6,NF- 魏 B, SHP1 and IFN- 尾 in PAM cells of PRRSV infected group and control group were quantitatively analyzed by real-time fluorescence quantitative PCR assay. The results showed that activated SHP1 promoted the transcription of IRF7,NF-kB and IFN- 尾 in PAM after PRRSV infection, and inhibited the transcription of IRF3 and TRAF6. LPS and Poly (I: C) stimulated PAM cells infected with PRRSV to promote IRF3,IRF7,TRAF6,. Transcription of NF- 魏 B, SHP1 and IFN- 尾. The results showed that activated SHP1 regulated the level of antiviral immune response induced by PRRSV through TLR mediated signaling pathway or unknown pathway. LPS and Poly (I / C) stimulated PRRSV infected PAM cells to enhance the transcription level of antiviral cytokines.
【作者單位】： 河南農(nóng)業(yè)大學(xué)牧醫(yī)工程學(xué)院;
【基金】：國家自然科學(xué)基金資助項目(31172346)
【分類號】：S852.65

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