| Viral pneumonia is one of the most common infectious diseases in pediatrics, Seriously threatened children’s health, pneumonia was listed as one of the four pediatric diseases by China’s Ministry of health.Viral pneumonia accounted for about half of child pneumonia. Respiratory syncytial virus (RSV) is one of the important pathogen of viral pneumonia in children, it is common in the viral pneumonia in children.At present, there is no specific anti RSV virus medicine and effective RSV vaccine in western medicine, while Chinese medicine has its unique advantages in treating viral infectious diseases. Clinical research and experiment research have proved Jinxin oral liquid is effective against RSV infection.TLRs as a type I transmembrane receptor, can identify the pathogenic molecules of different microbial correlation model.RSV is a single-stranded RNA virus, when it enters the body, ssRNA internal is exposed, and TLR7which can identify the virus ssRNA will play an important role.This study followed TLR7signal transduction pathway, investigated the regulatory role of Jinxin oral liquid on TLR7,IRF7and IFN-α induced by RSV infection, and explored the mechanism of Jinxin oral liquid anti RSV infection.ObjectiveTo study the role of Jinxin oral liquid on TLR7,IRF7and IFN-α induced by RSV, and explore its possible immunological mechanism in treating RSV pneumonia.MethodsThe expression levels of TLR7, IRF7and IFN-α were detected by Realtime RT-PC R,Western blot,ELSIA assay when RSV infected RAW264.7cells and BALB/c mice. Real time RT-PCR data were calculated by relative quantitative method, other data were statist ically analyzed with SPSS18.0software.ResultExperiments in vitro:1.24h after RAW264.7cells infected by RSV, the expression level of TLR7mRNA was higher than that in normal group(P<0.05; the expression levels of TLR7mRNA in both the ribavirin group and Jinxin oral liquid group were less than0.5times normal cells group, there is significant difference (P<0.05); compared with RSV infection group, the expression level of TLR7mRNA decreased (P<0.05), the Jin Xin medicated gelatin group down-regulation was more obvious than ribavirin. The expression level of IRF7mRNA was3.12times than the normal cells group, there is significant difference (P<0.05). The serum containing Jinxin oral liquid and the serum cotaining ribavirin down-regulated significantly the expression level of IRF7mRNA of RAW264.7cells infected by RSV(P<0.05),.2.24h after RAW264.7cells infected by RSV, compared with the normal group, the expression level of TLR7protein incrseaed, The serum containing Jinxin oral liquid down-regulated significantly the expression level of TLR7protein (P<0.01). The expression level of IRF7was higher than that in normal group (P<0.05), and compared with that in RSV group, the serum containing Jinxin oral liquid slightly increased the expression level of IRF7, but there was no statistical significance (P>0.05).3.24h after RAW264.7cells infected by RSV, in cell supernatants the expression of IFN-α very low.Experiments in vivo:1.24h after RSV infecting BALB/c mice, TLR7mRNA expression in lung tissue was significantly increased compared with the normal group (P>0.05). High-dosage and low-dosage Jinxin oral liquid prevention both down-regulated TLR7mRNA expression(P>0.05), and low-dosage Jinxin oral liquid prevention is obvious.72h after RSV infecting BALB/c mice, expression of TLR7mRNA decreased compared with that at24h, while that in Jin Xin group and ribavirin group significantly increased compared with that at24h, especially expression of TLR7mRNA in Jin Xin low-dosage treatment group is highest.144h after RSV infecting BALB/c mice, expression of TLR7mRNA declined further compared with that at72h, and expression of TLR7mRNA in Jin Xin group and ribavirin group decreased compared with that at144h,while increased compared with RSV group (P<0.05), especially expression of TLR7mRNA in Jin Xin high-dosage treatment group is highest.2.24h after RSV infecting BALB/c mice, TLR7mRNA expression in lung tissue was significantly increased compared with the normal group (P<0.01). while that in Jin Xin prevention group and ribavirin group significantly increased compared with that in RSV group(P<0.05), especially expression of IRF7mRNA in Jin Xin low-dosage prevention group is lowest.With lastingness of infection, expression of IRF7mRNA decreased gradually, while that in Jin Xin group and ribavirin group increased compared with that in RSV group, and that in Jin Xin high-dosage treatment group was higher than that in ribavirin group (p<0.05).3.Expression of TLR7after RSV infecting BALB/c mice(P<0.05), and Jinxin oral liquid down-regulated significantly the expression level of TLR7(P<0.05). During the early stages of infection, expression of IRF7in BALB/c in mice decreased, while Jinxin oral liquid down-regulated significantly the expression level of IRF7, during the late stage of infection, that in Jin Xin high-dosage treatment group was higher than that in RSV group (P<0.05).4.In this study, experiments in vivo show that express of IFN-a increased during the early stages of infection (P<0.05), while that in Jin Xin high-dosage treatment group and high-dosage treatment group was higher. With lastingness of infection, expression of IFN-a mRNA decreased gradually, while that in Jin Xin high-dosage treatment group and high-dosage treatment group increased significantly. Express of IFN-α decreased during the late stages of infection. Jinxin oral liquid down-regulated significantly the expression level of IFN-αcompared with that in RSV group(P<0.05).Conclusion1. Jinxin oral liquid could regulate the expression of TLR7and IRF7at DNA transcription and protein synthesis levels.2. Jinxin oral liquid could down-regulate the expression TLR7mRNA and IRF7mRNA in RAW264.7cells infected by RSV, and it could down-regulate the expression of TLR7protein,but it could barely down-regulate the expression of IRF7protein.3. Jinxin oral liquid could up-regulate the expressions of TLR7mRNA, IRF7mRNA and IRF7protein in BALB/c mice infected by RSV during the early stages of infection, it could up-regulate the expressions of during the late stages of infection, and it could down-regulate the expressions of TLR7proteinn.4. Jinxin oral liquid could up-regulate the expressions of IFN-α in BALB/c mice infected by RSV during the early stages of infection, while it could down-regulate the expression of IFN-α.5. Jinxin oral liquid is an effective drug in the treatment of RSV infection, and its mechanism is that it may regulate the expression of TLR7, IRF7, IFN-α in TLR7signal transduction pathway. |
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