| Objective:Based on the physiological characteristics of "Lung often inadequate " with Children,combine the basic theory of TCM of“strengthening the vital and eliminating the pathogen”and the treatment method,use vivo experimental techniques, study the treatment and prevention of pneumonia in young rats induced by the IV and dose-effect relationship used by double relieving superficies methods of“strengthening the vital and eliminating the pathogen”of influenza virus (IV) infection, discuss the effective mechanism of anti-influenza virus pneumonia from MYD88 dependent pathway and provide the experimental basis for the clinical treatment of IV.Materials and methods:1.the main activity of drugs active ingredients of the cream of detection rate as the evaluation process was obtained by orthogonal experiment and ethanol concentration, solvent ratio, extraction time and extraction times were used as the investigated factors, Index contents of quality control of the main constituents of Yu Ping Feng San and Yinqiaosan were used as the evaluation index to sieve and optimize the extraction artworks and to analysis their main active ingredient.2.The optimized artwork extracts of Yinqiaosan, Yupingfeng particle and particle of both on IV infection in vitro and dose-effect relationship was observed by Hemagglutination test.3.The pneumonia of young KM mouse induced by influenza virus through cavitas nasi inoculation, combined with the basic condition, pathological section of lung tissue, morphology and lung index of mouse to assess the IV pneumonia model of mouse to study the effectiveness of prevention and treatment IV pneumonia in mice and dose-effect relationship throuth double relieving superficies methods of“strengthening the vital and eliminating the pathogen”. 4.Used Immunohistochemical methods and RT-PCR method to detect TLR4, NF-?B P65 protein expression in lungs of mice to osserve the mechanism of prevention and treatment of Yinqiaosan extract, Yupingfeng particle and particle of both on IV pneumonia of mouse.5.Used Immunohistochemical methods and RT-PCR method to detect TLR4, NF-?B P65 protein expression in lungs of mice to osserve the mechanism of prevention and treatment at different time points with Yinqiaosan extract, Yupingfeng particle and particle of both on IV pneumonia of mouse.Results:1. the optimum extraction of Yupingfengsan is 70% ethanol concentration, solvent ratio of 10 times the amount of extracted twice, each time extraction for 2 hours; and the optimum extraction of Yinpingsan is 50% ethanol concentration, solvent multiples 10 times, extracted twice, each time extracting 3 hours.2. Yinqiaosan monarch drug extract, Yu Ping Feng San extracts, and Yinpingsan extracts infected chick embryos: prevention and treatment, there was no significant varieties of hemagglutinin valence between each concentrations and virus control group(P<0.05). compared with the virus control group, traditional Chinese medicine HA titer of each dose group were not statistically significant (P> 0.05). The dose medicine group was statistically significant (P> 0.05) ,except Yinqiaosan treatment of high concentration higher than the low concentration group.3. The prevention and treatment group could reduce the lung index of pneumonia in group IV mice, and Yinpingsan high-dose group, Yinqiaosan ,Yupingfengsan high dose group and high dose group IV pneumonia prevention and treatment of mice with no significant difference in the lung index ( P> 0.05); the Chinese high-dose group were decreased with Ribavirin IV pneumonia lung indices, no significant difference (P>0.05)except screens scattered high dose group. Except the low dose group Yinqiaosan Yupingfengsan lung index of less than low-dose group,the dose of the medicine between the two groups in the low dose group and between groups, IV pneumonia in the lower indices of the lungs of mice showed no significant difference ( P> 0.05).4.MYD88 protein, NF-?B P65 protein were higher than normal group for the prevention of the lung tissue of mice detected by Immunohistochemistry ; protein expression of C drug in high dose group, A drug in high dose group were lower than the other groups, comparison statistically significant (P <0.05). Followed by C drug in the dose group, B group of high-dose drug ribavirin group; prevention of drug of low dose group C, B drugs in the low dose group, A low dose of drug in the lung tissue of mice MYD88 group protein and model group There was no significant difference; prevention of drug in the low dose group A, B drugs in the low dose group, and C drugs in low dose group lung tissue NF-?B P65 protein expression and no significant difference between the model group.5.The prevention of the lung tissue of mice MYD88 mRNA, NF-?B P65 mRNA expression were higher than the normal group detected by RT-PCR method; C drug MYD88 mRNA expression in the high dose group were lower than the other treatment group, there were statistically significance (P <0.05); followed by high dose drug B, C drug in the dose group, no significant difference between the two groups. A low-dose drug group, B group and the model of low-dose drug group showed no significant difference. C drug in high dose group the expression of NF-?B P65mRNA was the lowest, followed by C drug in the dose group, A high dose of drug, B high dose drug and ribavirin control group, no statistical significance between groups. A low-dose drug group, B group and the model of low-dose drug group showed no significant difference.6.The expression of MYD88 protein, NF-?B P65 protein in lung tissue of mice in all treatment groups were higher than normal group tested by Immunohistochemistry; MyD88 of C drug dose group was the lowest, and other groups was statistically significant. followed by A drug in the high dose group, B drug in high dose group, C drug in the dose group, ribavirin control group, no significant difference between the two groups. NF-?B P65 protein expression of C drug in high dose group was the lowest, followed by drug A high dose, ribavirin group and C drugs in the dose group, but no statistical significance between groups (P <0.05); B drug low-dose group and model group was not statistically significant.7.The MYD88 mRNA, NF-?B P65 mRNA expression of the lung tissue of mice treated groups were higher than the normal group detected by RT-PCR method; MYD88 mRNA of C drug in high dose group was lower than other treatment group, there was statistically significance between groups. Followed by high dose group B drug, ribavirin group, A drug high dose group, C drug in the dose group, no statistical significance between groups. the expression of NF-?B P65mRNA of C & B high dose group were lowest, no significant difference between the two groups. Followed by C drug in the dose group, A drug high dose group, B drug in the dose group, ribavirin control group, no statistical significance between groups.8.at different times of double relieving superficies methods MyD88 protein expression in the lowest A treatment for the prevention of B group. A consolidation therapy followed by B, C treatment prevention of C, C consolidation treatment C, the group showed no statistical significance. NF-?B P65 protein A treatment for the prevention of the lowest in group B, followed by treatment for the prevention of C C group, treatment group A consolidation of B, the comparison between groups was not significant.9.at different times of double relieving superficies methods, MYD88 mRNA expression in the lowest consolidation for the treatment of C group consolidation of C, C treatment prevention of group C, treatment B A group consolidation, preventive treatment A group B, four groups were not significant. NF-?B P65 mRNA expression in the lowest treatment A group B for the prevention, prevention of C treatment C, the two groups was not significant. C followed by consolidation therapy group C, treatment B A group consolidation, the two groups was not significant.Conclusion:1.Two-table method has prevention and treatment on chick infected by IV.Yinqiaosan monarch drug extract, Yu Ping Feng San extracts, and yinpingsan extracts of different doses of IV infection of chicken have different levels of prevention and treatment of protection; the prevention and treatment of the Medicines is related to drug dose, but there was no significant difference between the dose.2. Two-table method has prevention and treatment on mice infected to pneumonia by IV.Yinqiaosan monarch drug extract, Yu Ping Feng San extracts, and Yinpingsan extracs have different doses t to reduce IV infection caused by inflammation of the lungs, reducing the role of lung injury ,and Yinpingsan high-dose group is the best.3. Its mechanism have a relationship with inhibition of MYD88-NF-?B P65 over-dependent signaling pathway activation.4. The two methods are used in combination at different times, preventing B drug and treating A drug, preventing C drug and treating C drug are bettermethod than others. |
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