The Research Of Immune Inflammatory Mechanism On The Viral Respiratory Infection Of Damp-heat Syndrome With HaoQinQingDan Decoction

The viral infection of respiratory tract is a common and frequently-occurring clinical disease,and all of the people are susceptible to it.Because of abounding rain water and hot climate in LingNan region of China,the viral respiratory infection have the characteristics of transforming into heat and intermingling dampness from differentiating syndrome,some researchers believe that damp-heat syndrome was embodied in the disease.Some heat-clearing and dampness-dispelling medicinal were widely used.HaoQinQingDan(HQQD) decoction is a Chinese herbal formula with the effect of heat-clearing and dampness-dispelling.The warm disease of dampness-heat syndrome of qi aspect is its indication.Based on some related clinical research,HQQD decoction was effective in the treatment of viral disease.It is practical significant to carry out the clinical and experimental research about the formula in viral respiratory infection of damp-heat syndrome.On the basis of clinical and experimental research,we explore the immune inflammatory mechanism of HQQD decoction for treating damp-heat syndrome through the methods of macro-observation and microscopic analysis.Partâ… Clinical ResearchObjective:To observe the effect of HQQD in the improving symptoms of TCM and the serum levels of tumor necrosis factor-α(TNF-α) and Interleukin-6(IL-6) of patients in treating viral upper respiratory infection of damp-heat syndrome.And to explore the clinical efficacy and the mechanism in the immune inflammatory.Methods:32 cases are selected and divided into two groups at random:HQQD group and Ribavirin group.Comparing the effect before and after the treatment interclass to know the effect of symptoms of damp-heat syndrome and the serum levels of TNF-αand IL-6 by the estimate tables of TCM symptom schedule and Enzyme-linked immunosorbent assay(ELISA).Results:There are no significant differences in sex,age and serious extent between HQQD group and Ribavirin group,so the two groups are comparable.It has statistical significance(P＜0.5) that both HQQD and Ribavirin can improve main and minor symptoms of damp-heat syndrome before and after treating.Seven main symptoms(fever,aversion to cold,heavy body and lassitude of the four limbs,oppression in the chest and gastric stuffiness,torpid intake and nausea and vomiting,sloppy stool,and dry mouth but not desire for drinking water.) and six minor symptoms(cough,pharyngeal itching pharyngodynia,nasal congestion,congestive throat,vertigo and head heaviness,and reddish scanty urine) of damp-heat syndrome can be relieved visibly after using HQQD(P＜0.05) than Ribavirin.Both HQQD and Ribavirin can decrease the serum levels of TNF-αand IL-6 of patients before and after treating(P＜0.05).The serum levels of TNF-αand IL-6 of patients are significant between HQQD group and Ribavirin group(P＜0.05).HQQD can decrease the serum levels better than Ribavirin.The indexes of laboratory for patients both HQQD group and Ribavirin group are normal after treating.HQQD is safety for patients.Conclusions:HQQD can not only improve the symptoms of viral upper respiratory infection of damp-heat syndrome but also decrease the serum levels of TNP-αand IL-6 for patients.HQQD is very effective in treating viral upper respiratory infection of damp-heat syndrome,and it may be its' mechanism suppressing the immune inflammatory.Partâ…¡Experimental research1.To explore the concentration of influenza A(H₁N₁) virus on mouse infected for experimental animal modelsObjective:Explore an appropriate virus infection dose of the production of animal models of damp-heat.Methods:The mice are randomly divided into eight groups and were given richly fatty and sweet diet and humid heat environment:dampness and heat group,seven groups with different concentrations(1 LD₅₀,5 LD₅₀,10 LD₅₀,20 LD₅₀,30 LD₅₀, 40 LD₅₀,50 LD₅₀).The symptoms,sign,survival rate of mice and pathogenic change of lung tissue were observed in the experimental process.Results:Dampness and heat group showed some features such as rapid gain weight, drinking more water,more plentiful excretion,etc.The mice infected influenza virus of clinical manifestations showed pneumonia.The symptoms and pathogenic change were closely related to virus infection.The survival rate of 1LD₅₀ group was the highest one.Conclusions:The appropriate virus infection dose of the production of animal models of damp-heat is 2LD₅₀.2.The research on animal models of damp-heat syndrome by adopting a richly fatty and sweet diet,humid heat environment and influenza virus infectedObjective:To produce animal models of damp-heat in the multiple factors.Methods:The mice were randomly divided into four groups,control group(A group),dampness and heat group(B group),model group of damp-heat syndrome (C group),and influenza virus group(D group).Analyzing and comparing death rate,lung index number,pathogenic change,expressions of influenza virus mRNA in the lung tissue,and expressions of cytokine TNF-α,IL-6 protein and mRNA in the lung tissue.Results:(1) Virus infection directly caused death.(2)The lung indexes number of C group or D group are higher than A group and B group(P＜0.05).There are not significant difference between A group and B group(P＞0.05),the same to between C group and D group(P＞0.05).(3)A group and B group show normal lung tissue,but pathogenic changes of C group and D group show interstitial pneumonia caused by virus infection.C group of mean rank is higher than D group for the degree of pathological changes.Ultrastructural observation: A group of lung tissue is normal;a large number of lipid droplets in the cytoplasm of alveolar epithelial are found in B group;C group show pathogenic changes of interstitial pneumonia and a large number of lipid droplets in the cytoplasm of alveolar epithelial are found;D group only show pathogenic changes of interstitial pneumonia.(4) Compared to D group,the expressions of influenza virus mRNA of C group increase in the lung tissue,but the difference is not significant(P＞0.05).(5) Compared to A group or B group, the expressions of cytokine TNF-αand IL-6 mRNA of C group and D group raise up in the lung tissue,and there are significant(P＜0.05).There are not significant difference between A group and B group(P＞0.05),the same to between C group and D group(P＞0.05).(6) Mean ranks of C group and D group are obviously higher than A group and B group for the expressions of cytokine TNF-αand IL-6 protein in the lung tissue.Mean rank of B group is higher than A group,and mean rank of C group is higher than D groupConclusions:Animal model of damp-heat syndrome have partial clinical characteristics of damp-heat syndrome,such as torpid intake,sloppy stool, etc.;but clinical manifestation show prominent pneumonia symptoms.Animal model have characteristics such as fat metabolism disorders,immune dysfunction,inflammatory factors increasing,etc.Internal and external humid heat environment may play an active role in the inflammatory cytokine secreted of TNF-αand IL-6.3.The research of the immune inflammatory mechanism of HQQD decoction on animal models of damp-heat syndromeObjective:The mice were treated by the different doses of HQQD in order to explore the immune inflammatory mechanism and find the most effective dose. Methods:The mice were randomly divided into six groups,control group(A group),model group of damp-heat syndrome(C group),Ribavirin group(E group), high,middle and low dose HQQD groups(F group,G group and H group).Analyzed and compared death rate,lung index number,pathogenic change,expressions of influenza virus mRNA in the lung tissue,and expressions of cytokine TNF-α, IL-6 protein and mRNA in the lung tissue.Results:(1)Each medicine can decrease the mortality rate of animals and the lung inflammatory reaction to some extent.(2)Compared with C group,the lung index number of E,F and G groups are significant difference except for H group (P＜0.05).The lung index number of F group is most obviously decreased. (3)Each medicine can decrease degree of pathological changes to some extent. We can surmise that the effect of high dose group is mostly obvious from the trend from the order of smallest mean rank.(4)Compared with C group,E,F, G and H groups are significant for the expressions of influenza virus mRNA in lung tissue(P＜0.05).The decrease of influenza virus may have a role in the mechanism of HQQD.The effect of E group is better than other groups,but the difference is not significant(P＞0.05).(5) Compared with C group,E, F and G groupsare significant for the expressions of cytokine TNF-αand IL-6 mRNA in lung tissue(P＜0.05).There are not significant difference between H group and C group(P＞0.05).HQQD can decrease the expressions of cytokine TNF-αand IL-6 mRNA.F group is obvious,but the difference is not significant (P＞0.05).(6)HQQD can decrease the expressions of cytokine TNF-αand IL-6 protein in the lung tissue to some extent.We can infer that the effect of F group is mostly obvious from the trend of mean rank order.Conclusions:HQQD may play an overall protection role in viral pneumonia of damp-heat syndrome of mouse,and decrease the expressions of cytokine TNF-αand IL-6 mRNA and protein in the lung tissue.The effect is closely related to dose,and high dose group is mostly obvious.4.The research of the immune inflammatory mechanism of HQQD prescription on animal models of damp-heat syndromeObjective:To explore the immune inflammatory mechanism of HQQD prescription according to the relation of formula of sovereign,minister,minister and courier medicinal.Methods:The mice were randomly divided into seven groups,control group(A group),model group of damp-heat syndrome(C group),high dose HQQD groups(F group),first group of prescription(I group),second group of prescription (J group),third group of prescription(K group),forth group of prescription (L group).Analyzed and compared death rate,lung index number,pathogenic change,expressions of influenza virus mRNA in the lung tissue,and expressions of cytokine TNF-α,IL-6 protein and mRNA in the lung tissue.Results:(1) Each TCM prescription can decrease the mortality rate of animals and the lung inflammatory reaction to some extent.(2)Compared with C group, the lung index number of F group is most obviously decreased,and it is better than I and J groups.(3)We infer that the effect of F group is mostly obvious in decreasing the degree of lung pathology from the mean rank trend.Group F is best,K and L groups are followed,but I and J groups are not obvious. (4)Compared with C group,HQQD prescription groups are significant for the expressions of influenza virus mRNA in lung tissue(P＜0.05).F group is better than other groups,but the difference is not significant(P＞0.05).The decrease of influenza virus may have a role in the mechanism of HQQD prescription.(5) Compared with C group,E,J,K and L groups are significant for the expressions of cytokine TNF-αand IL-6 mRNA in lung tissue(P＜0.05). There are not significant difference between I group and C group(P＞0.05). HQQD prescriptions can decrease the expressions of cytokine TNF-αand IL-6 mRNA.(6) HQQD prescriptions can decrease the expressions of cytokine TNF-αand IL-6 protein in the lung tissue to some extent.We can infer that the effect of F group is mostly obvious from the trend of mean rank order.Conclusions:HQQD prescription can decrease mortality rate of animals,lung inflammatory reaction,reproduction of influenza virus and expressions of cytokine TNF-αand IL-6 protein and mRNA in the lung tissue.The integrated HQQD prescription is the most effective.According to the results of HQQD prescription separated,sovereign medicinal(QingHao and HuangQin) is very important in the formula.Furthermore we infer that the mouse model of damp-heat syndrome have the characteristics of damp prevailing heat.