| Objective:Firstly, we are going to detect the mRNA levels of tumor necrosis factor alpha(TNF-α) in lung, SIgA in bronchoalveolar lavage fluid, intestinal fluid in rats with pneumonia induced by Respiratory Syncytial Virus(RSV), in order to reveal the molecular mechanisms of Jinxin Oral Liquid administrated through different ways on RSV.Secondly, we intend to observe the effect of serum containing Jinxin Oral Liquid, baicalin and emodin on the expression of TNF-αmRNA in A549 cells infected with RSV, so that we can evaluate the effect of Jinxin Oral Liquid and its effective monomer on the expression of TNF-αin the RSV infected rat model.Thirdly, we seek to investigate the effects of serum containing Jinxin Oral Liquid, baicalin and emodin against respiratory syncytial virus fusion in cells cultured in vitro by means of laser confocal scanning microscope.Methods:In this study, the experiments both in vitro and in vivo were conducted by means of experimental zoology, serum pharmacology, cell culture, Enzyme-linked immunosorbent assay and quantitative real-time PCR. Results were calculated by SPSS 10.0.Result:1. The expression of secreted immunoglobulin-A (SIgA) in bronchoalveolar lavage fluid and intestinal fluid in RSV pneumonia rat model were significantly higher than normal group as well as the level of TNF-αmRNA in lung tissue(P<0.01).2. The secretion of SIgA in bronchoalveolar lavage fluid and intestinal fluid of Jinxin Oral Liquid pretreated group, high-dosage of Jinxin Oral Liquid group and medium-dosage of Jinxin Oral Liquid group were significantly higher than the model group. The expression of TNF-αmRNA in lung tissue of Jinxin Oral Liquid pretreated group and medium-dosage of Jinxin Oral Liquid group were lower than the model group(P<0.05), in which the Jinxin Oral Liquid pretreated enema group reduced significantly (P<0.01).3. The secretion of SIgA in bronchoalveolar lavage fluid of Jinxin Oral Liquid pretreated group, high-dosage of Jinxin Oral Liquid group and medium-dosage of Jinxin Oral Liquid group were significantly higher than the ribavirin group(P<0.05 or P<0.01). The secretion of SIgA in intestinal fluid of high-dosage of Jinxin Oral Liquid was significantly higher than the ribavirin treated group(P<0.01), but the level of TNF-a mRNA in lung of the Jinxin Oral Liquid pretreated enema group were significantly lower than the ribavirin group(P<0.05)4. The secretion of SIgA in bronchoalveolar lavage fluid of pretreated group and high-dosage of Jinxin Oral Liquid group administrated orally were significantly higher than the Jinxin Oral Liquid group by clysis therapy. The secretion of SIgA in intestinal fluid of Jinxin Oral Liquid pretreated group administrated orally were significantly higher than the Jinxin Oral Liquid pretreated group by enteric administration, but the results in the secretion of SIgA in high-dosage of Jinxin Oral Liquid group were opposite. The level of TNF-a mRNA in lung tissue of pretreated group in Jinxin Oral Liquid group by enteric administration were significantly lower than high-dosage, medium-dosage and low-dosage of Jinxin Oral Liquid group administrated orally.5. After A549 cells were infected with RSV, the expression of TNF-a mRNA in the model group at 12h,24h and 48h time points respectively after the drug administration were significantly higher than the normal group.But the expression of TNF-a mRNA in the three groups(the blank serum group, the Jinxin Oral Liquid contained serum group, the ribavirin contained serum group) had no significant difference compared with the nomal group.And the expression of TNF-a mRNA in the emodin group, the baicalin group and in the both of emodin and baicalin group shows an earlier raised and later decreased state.The peak TNF-a mRNA of the emodin group were at 12h, respectively the baicalin group and the both of emodin and baicalin group 24h,and there were statistically significant differences compared with the model group.6. The results of confocal laser scanning microscopy (CLSM) indicated that the fluorescence were enhanced by RSV infected. In compared to the model group, the fluorescence in the emodin group and both the emodin and baicalin group decreased, and we did not find the obvious cell fusion. But there were more early apoptotic cells in the emodin group than others groups. There was no significant variance between the serum containing Jinxin Oral Liquid group and the model group in the fusion of RSV infected cell-to-cell.But there were less fusions in the serum containing Jinxin Oral Liquid group than the model group of RSV infected cell and the normal cell.There was no significant difference among the virus control group, the blank serum group, the ribavirin contained serum group and the model group in the fusion.Conclusion:1. Jinxin Oral Liquid were effective in treating pneumonia induced by RSV, and the effects were possibly mediated by modulate immune balance through down-regulating expression of TNF-a in the lung tissue and up-regulating expression of SIgA in respiratory mucosa..It can not only strengthen the immunity of the body, but also decrease local inflamative reaction.2. The retention-enema may be a available method in child who were infected with RSV, which deserve to be further investigated. 3. Emodin and baicalin can up-regulate the expression of TNF-a mRNA in RSV infected cells, besides, emodin was able to enhance cell apoptosis in infected cells, and may interfere with cell fusion as well.
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