摘要
目的:研究呼吸道合胞病毒(RSV)对细胞因子信号抑制因子(SOCS)3,Toll样受体(TLR)3、TLR4 mR-NA表达影响,探讨SOCS3在病毒感染上皮细胞后对Ⅰ/Ⅱ型细胞因子的调节效应。方法:设计人SOCS3特异小干扰RNA(si-RNA),转染上皮细胞A549,RSV感染后不同时段,逆转录聚合酶链反应(RT-PCR)半定量检测细胞不同时间点细胞SOCS3、TLR3、4 mRNA的表达,ELISA法检测培养上清中干扰素(IFN)-β和白细胞介素(IL)-8的浓度。结果:RSV感染上调A549细胞TLR3、4和SOCS3 mRNA的表达。SOCS3在感染后1 h升高,2 h达到峰值,分别为0 min的2.1倍和6.2倍(P<0.05),随后逐渐恢复至基础水平;TLR3、4在感染后2 h增加,并在24 h内持续表达较高水平。IL-8于24 h开始增高,24 h较基础水平增加25倍;IFN-β虽增高,但与基础水平无统计学差异。抑制SOCS3,对TLR3、TLR4 mRNA无显著性影响,但早期上调IFN-β的表达,IL-8虽有增高,但显著低于对照组。结论:RSV感染早期通过上调SOCS3表达调节上皮细胞差异表达Ⅰ/Ⅱ型细胞因子。
Objective： To explore the changes of SOCS3, TLR3, and TLR4 expressions and their effects on the regulation of type I / II cytokines of host in response to RSV infection. Methods： Human epithelia cells A549 was transfected with a specific siRNA against SOCS3, and the SOCS3 and TLR3/4 mRNA expressions were detected at different hours post-RSV infection （p. i. ） by using semiquantitative RT-PCR, and the INF-β and IL-8 concentrations in supernatants were checked by using ELISA method. Results： The expressions of SOCS3 and TLR3/4 mRNA increased in A549 cells infected RSV. The mRNA of SOCS3 began to increase at 1st hour and peaked at 2nd hour p. i. , thereafter, declined to the base level within 24 hours. TLR3/4 mRNA began to increase at 2nd hour p. i. and persisted at a high level in 24 hours p. i.. IL-8 began increasing at 24th hour p. i. dramatically, while IFN-β increased in trace amount, and demonstrated no significant difference from the base level. When SOCS3 was inhibited, IFN-β secretion increased in 24 hours, while IL-8 level was decreased as compared with the controls despite of its increasing as compared with the base level （P〈0.05）. There was no significant change in the expression of TLR3/4 in transfected cells after RSV infection. Conclusion： RSV regulates the differential dis- play of type I / II cytokines mediated by up-regulating the expression of SOCS3 in the early stage of viral infection.
出处
《武汉大学学报（医学版）》
CAS
北大核心
2010年第1期33-37,共5页
Medical Journal of Wuhan University
基金
国家自然科学基金资助项目(编号:30973220)
