肿瘤坏死因子受体相关因子6对卡波西肉瘤相关疱疹病毒感染后促炎因子的影响

Effect of tumor necrosis factor receptor-associated factor 6 on pro-inflammatory factors after Kaposi's sarcoma-associated herpesvirus infection

摘要:

目的 通过体外培养观察肿瘤坏死因子受体相关因子6(TRAF6)沉默和过表达后促炎因子白细胞介素(IL)-1α、IL-1β、IL-6、IL-8的表达,探索TRAF6对卡波西肉瘤相关疱疹病毒(KSHV)感染后促炎因子的影响,为KSHV致病机制的研究提供新思路。方法 使用KSHV(+)iSLK细胞系建立沉默和过表达TRAF6模型,实时荧光定量聚合酶链式反应(RT-PCR)检测转染水平。细胞培养48、72 h后通过酶联免疫吸附试验(ELISA)分别检测促炎因子的表达,并比较TRAF6沉默及过表达后促炎因子表达水平的变化。结果 成功建立TRAF6沉默(siTRAF6)和TRAF6过表达(TRAF6-OE)细胞模型。ELISA结果显示,TRAF6沉默后促炎因子IL-1α、IL-1β、IL-6、IL-8表现出不同程度的升高,TRAF6过表达后IL-1α、IL-1β、IL-6、IL-8有不同程度的降低,且IL-6、IL-8变化幅度较大,说明TRAF6对促炎因子有一定的抑制作用,提示其在KSHV感染后促炎作用的信号通路中发挥重要作用。结论 TRAF6对促炎因子有一定抑制作用,有望作为新的潜在靶点抑制卡波西肉瘤病情发展。

Abstract:

Objective To observe the expression of pro-inflammatory factors including interleukin (IL)-1α, IL-1β, IL-6 and IL-8 after silencing and over-expressing of tumor necrosis factor receptor-associated factor 6 (TRAF6) in vitro, explore the effect of TRAF6 on pro-inflammatory factors after Kaposi’s sarcoma-associated herpesvirus (KSHV) infection, and provide new ideas for the study of the pathogenesis of KSHV. Methods The silence and over-expression TRAF6 models were established using KSHV(+)iSLK cell line, and the transfection level was detected by real-time fluorescence quantitative polymerase chain reaction (RT-PCR). After 48 and 72 hours of cultivation of cells, the expression levels of pro-inflammatory factors after silencing and over-expressing TRAF6 were detected by enzyme-linked immunosorbent assay (ELISA) and compared. Results TRAF6-silence (siTRAF6) and TRAF6-over-expression (TRAF6-OE) cell models were established successfully. ELISA results showed that pro-inflammatory factors IL-1α, IL-1β, IL-6, and IL-8 increased in varying degrees after silencing TRAF6, while these pro-inflammatory factors decreased in varying degrees after TRAF6 over-expression, IL-6 and IL-8 levels changed remarkablely, suggesting that TRAF6 had a certain inhibitory effect on pro-inflammatory factors and played an important role in the signaling pathway of pro-inflammatory effects after KSHV infection. Conclusion TRAF6 has a certain inhibitory effect on pro-inflammatory factors and is expected to serve as a potential target to inhibit the progression of Kaposi’s sarcoma.