重组腺病毒介导的sTLR2和sTLR4体外抗炎效应的实验研究

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。

Experimental study on anti-inflammatory efficacy of adenovirus-mediated sTLR2 and sTLR4

This study aimed to explore the effects of recombinated adenovirus-mediated sTLR2 and sTLR4 on the expression of inflammatory cytokines.Human monocytes THP-1 were divided into five groups,including negative control group,positive control group,AdTLR2 group,AdTLR4 group,and AdTLR2+AdTLR4 group.RT-PCR was carried out to detect the expression levels of TLR2 mRNA and TLR4 mRNA after cells were treated with recombinated AdTLR2 or recombinated AdTLR4 or the combination of AdTLR2 and AdTLR4.The results showed that high expression levels of TLR2 and TLR4 were achieved after THP-1 cells were transfected with the combination of AdTLR2 and AdTLR4.In contrast to the positive control,the levels of TNF-α,IL-1β,IL-6,IL-10,and IL-13 in inflammatory cells treated with AdTLR2 or AdTLR4 or the combination of AdTLR2 and AdTLR4 were notably reduced.Furthermore,these cytokines in AdTLR2+AdTLR4 group were further decreased compared with those in AdTLR2 group or AdTLR4 group(P < 0.05).In conclusion,the recombinated adenovirus AdTLR2 and AdTLR4 could secrete sTLR2 and sTLR4,thus inhibit inflammatory cell activation and cytokine release.