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目的观察重组腺病毒介导的sTLR2和sTLR4对炎症模型细胞因子表达水平的影响。方法将人单核细胞系THP-1细胞随机分为正常对照组、炎症对照组、重组腺病毒AdTLR2组、重组腺病毒AdTLR4组、重组腺病毒AdTLR2+AdTLR4组。AdTLR2和AdTLR4单独或者联合作用于各组细胞,通过RT-PCR方法检测TLR2和TLR4 mRNA表达,应用ELISA法对上清液中细胞因子TNF-α、IL-1β、IL-6、IL-10和IL-13的含量进行检测,探讨AdTLR2和AdTLR4对LPS诱导的THP-1细胞上清细胞因子表达水平的影响。结果通过RT-PCR方法检测THP-1细胞转染AdTLR2+AdTLR4后,TLR2和TLR4能够获得高表达。AdTLR2和AdTLR4单独和联合作用于LPS所致炎症细胞后,细胞上清中TNF-α、IL-1β、IL-6、IL-10和IL-13的含量较炎症对照组显著降低,且联合作用较单独作用组进一步降低,具有统计学意义(P<0.05)。结论重组腺病毒AdTLR2和AdTLR4能够降低LPS所致炎性细胞上清中TNF-α、IL-1β、IL-6、IL-10和IL-13的含量,并且联合使用的抗炎效果优于AdTLR2或AdTLR4的单独作用。提示制备的重组腺病毒能够分泌sTLR2和sTLR4,能有效抑制炎性细胞的活化,减少细胞因子的释放,具有一定的抗炎作用。
Objective To observe the effect of sTLR2 and sTLR4 mediated by recombinant adenovirus on the expression of cytokines in inflammatory model. Methods Human monocytic THP-1 cells were randomly divided into normal control group, inflammatory control group, recombinant adenovirus AdTLR2 group, recombinant adenovirus AdTLR4 group and recombinant adenovirus AdTLR2 + AdTLR4 group. AdTLR2 and AdTLR4 alone or in combination with each group of cells, by RT-PCR method to detect the expression of TLR2 and TLR4 mRNA in the supernatant by ELISA method of cytokines TNF-α, IL-1β, IL-6, IL-10 and IL-13 levels were detected to investigate AdTLR2 and AdTLR4 LPS-induced THP-1 cells supernatant cytokine expression levels. Results The expression of TLR2 and TLR4 in THP-1 cells transfected with AdTLR2 + AdTLR4 was detected by RT-PCR. The levels of TNF-α, IL-1β, IL-6, IL-10 and IL-13 in the supernatant of the LPS-treated mice were significantly lower than those in the control group by AdTLR2 and AdTLR4 alone and in combination Compared with the single effect group to further reduce, with statistical significance (P <0.05). Conclusion The recombinant adenovirus AdTLR2 and AdTLR4 can reduce the levels of TNF-α, IL-1β, IL-6, IL-10 and IL-13 in the supernatant of inflammatory cells induced by LPS, Or AdTLR4 alone. It is suggested that the recombinant adenovirus can secrete sTLR2 and sTLR4, which can effectively inhibit the activation of inflammatory cells and reduce the release of cytokines, and have a certain anti-inflammatory effect.