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目的检测汉滩病毒感染血管内皮细胞后IL-6,IL-8和TNF-α的分泌变化及其与TLR4的关系。方法用5LgTCID50/mL的HTNV76-1180.2mL感染EVC-304细胞(TLR4+)和EVC-304TS4(TLR4-)分别为实验组,以病毒未感染为阴性对照组,以LPS(2μg/mL)刺激作为阳性对照。48h后取细胞培养上清,用人IL-6,IL-8和TNF-α定量EIA试剂盒分别检测IL-6,IL-8和TNF-α在两个细胞系感染前后的分泌水平。结果IL-8在两个细胞系中感染前后的变化不明显,IL-6和TNF-α在EVC-304细胞系中,HTNV感染后升高,而在TLR4表达阴性的EVC-304细胞中,感染前后变化不明显。结论在TLR4表达阳性的EVC-304细胞中IL-6和TNF-α分泌增加,血管内皮细胞EVC-304在HTNV感染后的IL-6和TNF-α分泌可能是TLR4介导的。
Objective To detect the secretion of IL-6, IL-8 and TNF-α after Hantaan virus infected vascular endothelial cells and its relationship with TLR4. Methods Infection of EVC-304 cells (TLR4 +) and EVC-304TS4 (TLR4-) with 5Lg TCID50 / mL of HTNV76-1180.2mL were used as the experimental group. The uninfected virus was negative control group and stimulated with LPS (2μg / mL) Control. After 48h, the cell culture supernatants were taken and the secretion levels of IL-6, IL-8 and TNF-α were measured before and after infection by using IL-6, IL-8 and TNF-α quantitative EIA kit respectively. Results The changes of IL-8 in both cell lines were not obvious before and after infection. The expression of IL-6 and TNF-α in EVC-304 cell line increased after infection with HTNV. In EVC-304 cells with negative TLR4 expression, Before and after infection did not change significantly. Conclusions The secretion of IL-6 and TNF-α in EVC-304 cells with positive TLR4 expression is increased, and the secretion of IL-6 and TNF-α in EVC-304 cells after HTNV infection may be TLR4-mediated.