Effect Of Jinxin Oral Liquid On Interfering TLR4Expression On TRIF/NF-κB Activated By RSV

Instructor led by Mentor professor Professor Wang Shouchuan developed Jinxin oral liquid been confirmed in preliminary clinical and experimental research is an effective anti-RSV infection traditional Chinese medicine compound.The issue preliminary research has indicated Jin Xin oral liquid can regulate RSV infection induced the TLR4activation and the expression level of downstream signaling molecules. Given the multi-target effects of Chinese medicine, this experiment from the interference of TLR4expression, discusses Jin Xin oral anti-RSV infection in oral liquid regulation of TLR4signaling pathway, and the correlation with other TLRs.ObjectiveThe purpose of this experimental research is to interfere with the expression of TLR4, Jin Xin oral on the influence of TLR4signaling pathways downstream of related proteins, aims to further elucidate Jinxin oral pediatric respiratory syncytial virus pneumonia by immunomodulatory therapy.MethodsIn this study, the use of the small RNA interference technology TLR4-siRNA liposome and transfection Lipofectamine2000complex RAW264.7cells, Realtime-PCR method to detect TLR4mRAN transcription, determine the efficiency of siRNA.RAW264.7cells after TLR4expression is interference, to RSV infection, and then treated using Jin Xin oral containing medicine serum and collecting cells, Realtime RT-PCR, Western blot assay to detect TLR4and key molecular signaling pathways TRIF, NF-kappa B mRNA and protein expression.Result1. In vitro chemical synthesis of TLR4siRNA, transfected by liposome Lipofectamine2000RAW264.7cells, can effectively interfere with the expression of TLR4and its mRNA expression inhibition rate of57.4%(P<0.05).2. After the interference of TLR4expression, the expression of TLR4signaling pathways downstream signaling molecule NF-κB mRNA and protein were also lower (expression inhibition rate of25.8%)(P<0.05). But, the inhibition rate lower than TLR4, showed that the expression of NF-κB is also regulated by other TLRs, which shows that TLR4signaling pathway is just one pathway after RSV infection.3. After the interference of TLR4expression, TRIF mRNA and protein expression was significantly increased (expression increased1.835times)(P>0.05). Indicate the presence of negative regulation between TLR4and TRIF relationship may also synergies with TLR3. Meanwhile Description RSV infection, may activate TLR4/TRIF/NF-κB signaling pathway 4. Jinxin oral liquid can increase after RSV infection of TLR4, NF-κB mRNA expression and protein (P<0.05), but TRIF mRNA and protein expression was no significant regulatory effect (P>0.05).Conclusion1. TLR4siRNA can effectively interfere with the expression of TLR42. Jinxin oral liquid are effective RSV infection,Its role not only by adjusting the RSV induced TLR4/NF-κB signaling pathways, but also rely on other TLRs signaling pathways, but not after TLR4/TRIF way.And Jin Xin oral liquid also through other pathway regulating downstream signaling molecules generated, reflecting the multiple target effect of Chinese medicine compound function.