Toxic Hot Injection Studies On The Mechanism Of Influenza Virus Pneumonia In Mice Treated With

Influenza viruses are important pathogens in respiratory tract infections and periodic worldwide epidemic, posing a serious threat to human's life and health. Chinese medicine, which has a unique effect in prevention and treatment of viruses infection,little drug side effects,rich source of low drug cost and other advantages to improve the clinical symptoms, has shown the unique advantages in the treatment of Influenza viral infection. Therefore, a really good job in anti-influenza pneumonia medicine research is of great significance.In this experiment, we in-depth study the regulation mechanism of Dureping injection after influenza virus infection on the lung tissue level, especially from the level of gene and cytokine levels so as to provide a theoretical basis for clinical application.OBJECTIVE:Dureping Injection comprised of scutellaria,cape jasmine fruit,pig gall, fleabane flower, is a TCM compound preparation. In the experiment, we study the role of Dureping injection on mice Influenza viruse pneumonia of AP-1 gene in the lung tissue cells and the three cytokines MMP-1, MMP-9,TIMP-1 in the lung tissue, to analyze influenza viruses activate target genes and secrete cytokine to mediate inflammatory response and the mechanism of lung injury, providing theoretical basis for the traditional Chinese medicines used in clinical treatment of influenza virus pneumonia.METHODS:1. The effect of Dureping Injection on the lung index and the pathological change in mice with influenza virus pneumonia66 male ICR mice were randomly divided into the normal group, the model group, Dureping Injection of low-dose group, Dureping Injection of medium-dose group, Dureping Injection of high-dose group and positive group of Ribavirin,11 in each group. The pneumonia of mice with influenza virus infection model was built by using Influenza virus strain FM1(1/3LD50). Mice were injected intraperitoneally with 0.3ml of injections everyday, respectively.After 5-day treatment, the mice were killed to obtain the lung index and routine HE stained sections were examined. 2. The effect of Dureping Injection on the transcription factor AP-1 gene in the lung tissue cells of mice with influenza virus pneumoniaAnimal grouping, modeling, drug delivery methods ibid. After 5-day treatment, the mice were killed to obtain the lung tissue and c-jun and c-fos mRNA expression in lung tissues was detected by RT-PCR. 3. The effect of Dureping Injection on MMP-1,MMP-9 and TIMP-1 in the lung tissue of mice with influenza virus pneumoniaAnimal grouping, modeling, drug delivery methods ibid. After 5-day treatment, the mice were killed to obtain the lung tissue. After weight the lung tissue, we take it into a glass homogenizer, in accordance with the proportion of 1:9 by adding cold saline quality; then make the lung tissue to be homogenized fully. The prepared homogenate to 3000r/min speed centrifuge, extracting the supernatant. The content of MMP-1,MMP-9 and TIMP-1 in lung tissue were detected by ELISA double antibody sandwich Method.RESULTS:1.The effect of Dureping Injection on the lung index and the pathological change in mice with influenza virus pneumoniaMice with pneumonia of FM1 influenza virus infection, The lung index in model group were increased than in normal group (P<0.01); Compared with the model group, each group of Dureping Injection were lower, showing the role of inhibition of lung inflammation; the effect of medium-dose group is better(Inhibition percentage is 26.35%). HE slices staining observed by light microscopy, the pathomorphology studies showed that there are some similar alveoli in normal group, the thickness of alveolar wall are consistent and the inflammatory cells were not saw. The model group lung tissue showed a serious interstitial pneumonia changes. The normal structure of lung tissue such as alveoli, alveoli ducts and alveolar interval were changed; many lymphocytes infiltrate into the alveolar wall. Compared with Model group, the pathological changes of DRP group and Ribavirin group were ameliorated significantly. 2.The effect of Dureping Injection on the transcription factor AP-1 gene in the lung tissue cells of mice with influenza virus pneumoniaMice with pneumonia of FM1 influenza virus infection, The mRNA content of c-jun and c-fos in lung tissue cells in model group were increased than in normal group; Compared with the model group, the mRNA content of c-jun in the Dureping Injection of low-dose and medium-dose group were lower (P<0.05); the mRNA content of c-fos in each group were reduced (P<0.05). 3. The effect of Dureping Injection on MMP-1,MMP-9 and TIMP-1 in the lung tissue of mice with influenza virus pneumoniaMice with pneumonia of FM1 influenza virus infection, compared with normal group, the content of MMP-1 in lung tissue were enhanced(P<0.05); the content of MMP-9 in lung tissue were increased significantly in model group than in normal group(P<0.01), while no significant difference was found at the TIMP-1(P> 0.05). Compared with the model group, the content of MMP-1 in all group of Dureping Injection were lower; the content of MMP-9 in low-dose group and medium-dose group of Dureping Injection were significantly lower(P<0.01); the level of TIMP-1 in all groups of the Injection were significantly higher(P<0.01).CONCLUSION:1.Mice infected with influenza virus FM1 cause significant pathological lung injury, lung index was significantly higher; Dureping Injection can reduce the injury of lung inflammation, decrease the lung index so as to improve the lung pathological changes.2.Mice infected with influenza virus FM1, the AP-1 gene expression was enhanced, while Dureping Injection can decrease the level of AP-1 to reduce the release of inflammatory mediators.3.Dureping Injection can decrease the content of MMP-1 and MMP-9, increase the level of TIMP-1 in lung tissue to restrain inflammation.