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目的:通过研究金欣口服液含药血清对呼吸道合胞病毒(RSV)活化诱导的Toll样受体3(TLR3)的干预作用,探讨其治疗RSV肺炎可能的免疫学机制。方法:RSV感染体外培养的RAW264.7细胞,采用金欣口服液含药血清进行干预,并设利巴韦林阳性对照,24h后收集细胞,Real time RT-PCR法测定TLR3 mRNA的表达变化;激光共聚焦显微镜观察免疫荧光细胞化学染色法处理的各组细胞TLR3表达水平;ELISA法检测各组细胞培养上清中白细胞介素-6(IL-6)含量,在核酸及蛋白水平探讨金欣口服液含药血清抗RSV感染的作用机制。结果:RSV感染RAW264.7细胞后24h,细胞中TLR3 mRNA和蛋白表达水平以及细胞培养上清中IL-6表达明显升高(P<0.01),而金欣组TLR3及IL-6表达均显著低于RSV感染组(P<0.01,P<0.05),且效果优于利巴韦林组(P<0.01,P<0.05)。结论:金欣口服液含药血清能下调RSV活化诱导的TLR3及其下游炎症因子IL-6的高表达,推测其为金欣口服液抗RSV感染的机制之一。
OBJECTIVE: To investigate the possible immunological mechanism of treatment of RSV pneumonia by studying the intervention effect of Jinxin oral liquid containing serum on Toll-like receptor 3 (TLR3) induced by respiratory syncytial virus (RSV) activation. Methods: RAW264.7 cells cultured in vitro were infected with RSV. Jinli oral liquid serum was used for intervention. Positive control of ribavirin was established. Cells were collected after 24 hours. Real-time RT-PCR was used to detect the expression of TLR3 mRNA. The expression of TLR3 was detected by confocal laser scanning microscopy (FCM). The levels of IL-6 in the culture supernatants were detected by ELISA. The levels of nucleic acid and protein were determined by ELISA The Mechanism of Anti-RSV Infection in Oral Liquid Containing Serum. Results: The expression of TLR3 mRNA and protein and the expression of IL-6 in cell culture supernatant of RSV-infected RAW264.7 cells were significantly increased (P <0.01) 24 h after RSV infection, while the expressions of TLR3 and IL-6 in Jinxin group were significantly (P <0.01, P <0.05), and the effect was better than ribavirin group (P <0.01, P <0.05). CONCLUSION: Jinxin oral liquid-containing serum can down-regulate the expression of TLR3 and its downstream inflammatory factor IL-6 induced by RSV activation, suggesting that it is one of the mechanisms of Jinxin oral liquid anti-RSV infection.