| Viral pneumonia(VP)is one of the most common respiratory infectious diseases in clinic.It is common in children,the elderly,immunocompromised hosts and patients with heart and lung diseases.In recent years,with the accelerated aging of society and increase the host immune injury,virus mutation and the generation of new viruses,including respiratory syncytial virus(RSV)pneumonia,the incidence of VP was significantly increased,a serious threat to human health.At present,the treatment is still lack of strong specific therapeutic agents.Traditional Chinese medicine is characterized by holistic concept and syndrome differentiation and treatment.It has the characteristics of multi targets,multiple pathways and many links.It has obvious advantages in the prevention and treatment of viral infectious diseases.My doctoral supervisor certificate for decades with the research of traditional Chinese medicine in prevention and treatment of respiratory tract infectious diseases for long-term.In his opinion,deficiency,poison,phlegm and blood stasis is the the key pathogenesis of viral pneumonia,and treatment should be based on Strengthening Qi,detoxifying and removing blood stasis.Then he formulated the Fuzheng Jiedu Huayu Recipe.long-term practice proved that safe and reliable.The side from ginseng,Scutellaria,forsythia,radix paeoniae rubra,uniflower swisscentaury root,Trichosanthes,Patrinia and coix seed,has a rousing,detoxification,blood circulation function,mainly used for the treatment of pulmonary infection caused by multiple pathogens.Previous studies have shown that Fuzheng Jiedu Huayu Recipe can effectively alleviate pneumonia symptoms,promote inflammation absorption and control disease progression.The effect of lung infection caused by RSV is remarkable,but the material basis and mechanism of the treatment are not clear.In this study,RSV pneumonia was selected as the breakthrough point to investigate the mechanism of Fuzheng Jiedu Huayu Recipe and its effective fraction in the intervention of viral pneumonia.This thesis consists of two parts:summary and experimental research.In the review part,the understanding and research progress of TCM and modern medicine on RSV pneumonia were systematically expounded,which provided a theoretical basis for experimental research.In the experimental part,the mechanism of antiviral action of Fuzheng Jiedu Huayu Recipe and its effective fraction was investigated mainly from the aspects of morphology and cell biology.Objective:To investigate the mechanism of Fuzheng Jiedu Huayu Recipe and its effective fraction in RSV pneumonia mice from apoptosis gene,PI3K/Akt pathway related regulatory protein,cyto-kines and lung tissue pathological changes.Methods:180 female BALB/c mice were randomly divided into 3 groups at the time of first,3 and 5 days after infection,with 60 mice in each group.Each group was divided into 6 groups:normal group,model group,whole Party group,total glycoside group,volatile oil group and lymph node group,with a total of 10 mice in each group.BALB/c mice with RSV nasal infection were treated with Fuzheng Jiedu Huayu Recipe and its effective fraction,and the lung tissue was calculated at first,3 and 5 days after the first infection.The pathological changes of lung tissue were observed by optical microscope and electron microscope.The expression of Fas,P53 and caspase-12 in lung tissue was detected by Real-time PCR.Immunohistochemical method was used to detect the expression of AKT,GSK-3 and NF-kappa p65 protein in lung tissue of mice.ELISA method was used to detect the expression of IL-1,IL-8,IFN-,TLR3 and TLR7 in lung tissue.Results:1.RSV infected BALB/c mice lung indexAt different time points after infection,the lung index of the model group increased and showed an increasing trend.Compared with the normal group,there were significant differences(P<0.01).At first,3,and 5 days after infection,the lung index was significantly reduced in each administration group,and there was significant difference Compared with the model group(P<0.05).On the third and 5 day after infection,the effect of the whole group and Leigh Bhave Lin group was better than that of the total glycosides group and the volatile oil group(P<0.05).The inhibition effect of the whole group on the lung index was better than that of the total glycosides group and the volatile oil group,the effect is obvious at third days after infection.2.Pathological changes of lung in RSV infected BALB/c miceThe main manifestations of RSV infection were pulmonary interstitial inflammation,congestion,edema and thickening of alveolar wall,infiltration of inflammatory cells in the pulmonary interstitium,rupture of alveolar wall and enlargement of alveolar cavity.Under electron microscope,type ? alveolar epithelial cell swelling,containing a large number of pinocytotic vesicles,endothelial structure is not complete;type ? alveolar cell enlargement,filled with round or oval large number of virus particles in the nucleus,the structure of fuzzy;cytoplasmic organelles disintegrated,endoplasmic reticulum,mitochondria,ribosome,blurred.Pulmonary lesions were more severe in third and 5 days after infection.At different time points after infection,the degree of lung lesions in the prescription of Fuzheng Jiedu Huayu Decoction and the effective position group were reduced to varying degrees.3.Expression of apoptosis genes in lung tissues of RSV infected BALB/c miceThe first and 3 days after infection,compared with the model group,the treatment group can reduce the expression of Fas in the lung tissue of mice,in which the total glycosides and volatile oil group down significantly(P<0.01).First day after infection,the model group and drug group P53 expression were significantly increased(P<0.01);the expression of whole group,total glycosides group and volatile oil group were significantly reduced in the lung tissues of P53 and caspase-12(P<0.01),the total glycosides group and volatile oil group decreased obviously.On the third day after infection,the expression of caspase-12 was decreased significantly in the total glycosides group and the volatile oil group(P<0.05).4.Expression of PI3K/Akt related protein in lung tissue of RSV infected BALB/c miceThird days after RSV infection,compared with the model group,the expression of AKT in the lung tissue of the whole group increased significantly(P<0.01),but decreased significantly at the end of the fifth day.At third and 5 days after infection,the expression of GSK-3 was significantly higher in the whole group and the total glycosides group(P<0.01),which was significantly lower than that in the first day.The expression of NF-kB was significantly decreased in the whole group at first and 5 days after infection,but in the essential oil group,the expression of P65 was significantly decreased at the first day(P<0.05).5.Expression of cytokines in lung tissue of RSV infected BALB/c miceAt different time points after infection,the expression of IL-8 in lung tissue of model group was higher than that of normal group,and the first days is obvious(P<0.01);total glycosides group and volatile oil group can obviously down regulate the expression level of IL-8(P<0.05).The expression of IL-1 in each group was highest in first days after infection,with the passage of time,there is a downward trend;first day after infection,the whole party,total glycosides and volatile oil group can decrease the expression of IL-1(P<0.01);total glycosides were still significantly lower in third days.On the first day after infection,compared with the model group,the expression of IFN-was up-regulated in the three Chinese medicine groups,but there was no significant difference.The expression of IFN-was significantly increased in the whole group(P<0.05)after the infection for the last third days.For TLR7,only first days after infection,the model group,the total glycosides group and the volatile oil group were significantly higher than those in the normal group(P<0.05).In the whole process of RSV infection,Fuzheng Jiedu Huayu Decoction and its effective parts had no obvious effect on the expression of TLR3.Conclusion:1 Fuzheng Jiedu Huayu Recipe and its effective fraction have a significant inhibitory effect on the lung index of RSV infected BALB/c mice;2 Fuzheng Jiedu Huayu Recipe and its effective fraction can improve the lung lesions of RSV infected BALB/c mice;3 the expression of Fas,P53 and caspase-12 could be down regulated by Fuzheng Jiedu Huayu Recipe and its effective fraction;4 the effects of Fuzheng Jiedu Huayu Recipe and its effective fraction on PI3K/Akt pathway related proteins Akt,GSK-3 beta and NF-kappa B show different characteristics at different time after infection.5 Fuzheng Jiedu Huayu Recipe and its effective fraction can significantly down regulate the expression of IL-1,IL-8,and up regulate the expression of IFN-a,and have no obvious effect on the expression of TLR3 and TLR7.6 Fuzheng Jiedu Huayu Recipe and its effective fraction is an effective drug for treatment of RSV infection,its mechanism may be achieved by regulating the expression of apoptosis gene,PI3K/Akt pathway related proteins and many cytokines. |
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