| Background:Damp-heat syndrome of seasonal febrile disease was common clinical disease,caused by exogenous damp-heat factor and internal damage of the spleen and stomach.Because exogenous damp-heat factor easily lead to internal damage of the spleen and stomach, damp-heat in middle-JIAO or splenogastric hygropyrexia was typical phase.Pathogenic mechanism of damp-heat syndrome of seasonal febrile disease was not limited to one entrails or target,it was relate to ETX,lipid level,body fluid metabolism,inflammatory cell factor,immunologic function and gut hormone etc.Because of abounding rain water and hot climate in Guangdong province,damp-heat disease was common in clinic.Decoction of Hao qin qing dan,Coptidis Decoction for Detoxification and Three-seeds Decoction were generally apply to treat the damp-heat syndrome.It was important significance to study damp-heat syndrome in Guangdong province.It was accepted to establish models of damp-heat syndrome by adopting a richly fatty and sweet diet,humid heat environment and infecting by the biology causative agent.Biology causative agent was concentrated in Bacillus coli,Salmonella typhimurium and ETX.Different biology causative agent could lead to variant kind of Damp-heat seasonal febrile disease.The model established by virus factor was supplementary for studying Damp-heat syndrome of seasonal febrile disease.Objective:To study pathogenic mechanism of damp-heat syndrome of seasonal febrile disease in various component element by establishing mouse models of Damp-heat syndrome of seasonal febrile disease infected by MHV—A59 and Bacillus coli.To study pathogenic position of etiological factor by analyzing composite factor(richly fatty and sweet diet + humid heat environment + infecting by MHV—A59),humid heat factor(richly fatty and sweet diet + humid heat environment)and virus factor(infecting by MHV—A59).To certificate the pathogenic mechanism of damp-heat syndrome of seasonal febrile disease by treating this damp-heat syndrome infected by MHV—A59 with Decoction of Hao qin qing dan,Coptidis Decoction for Detoxification and Three-seeds Decoction,observe mechanism of action of therapy of clearing away heat and dampness.Disease caused by exogenous damp-heat factor was representative of damp-heat disease.It was supplementary and evidence of pathogenic mechanism of damp-heat syndrome of seasonal febrile disease to study immune level of peripheral blood of patients of exogenous damp-heat disease.Methods:1.literature research:on the basis of the approach of evaluation approach and review articles,we proposed to arrange study state of animal model of damp-heat syndrome of seasonal febrile disease,analyze study insufficiency of pathogenic mechanism of damp-heat syndrome of seasonal febrile disease,sum up study progression of MHV and the therapy of cleating away heat and dampness.2.Empirical research:①four-weeks-old male mouse of BALB/c was divided into three groups randomly,control group,model group(richly fatty and sweet diet + humid heat environment + infecting by MHV-A59),Bacillus coli group(richly fatty and sweet diet + humid heat environment + infecting by Bacillus coli).The TG and TNF-αlevel in serum of mouse were detected by biochemistry and ELISA.In experiment we observed the symptom and sign of mouse.②pathogenic mechanism of damp-heat syndrome of seasonal febrile disease:four-weeks-old mouse of BALB/c was divided into three groups randomly,control group,model group,Bacillus coli group.The level of IL-4 and IFN-γin mouse serum were detected by ELISA.The expression of NF-κB in mouse liver tissue and AQP4 in gastric mucous membrane was analyzed by immunohistochemistry.The percentage of T lympholeukocyte subset in peripheral blood of mouse was analyzed by flow cytometry.③Analyze the position of etiological factor in damp-heat syndrome of seasonal febrile disease:four-weeks-old mouse of BALB/c mouse was.divided into four groups randomly, control group,model group,Bacillus coli group,damp-heat group(richly fatty and sweet diet + humid heat environment),MHV-A59 group(infecting by MHV—A59).The quantity of MHV-A59 in mouse liver tissue was detected by fluorescent quantitation PCR,The level of IL-4 and IFN-γin mouse serum were detected by ELISA.The expression of NF-κB in mouse liver tissue and AQP4 in gastric mucous membrane was analyzed by immunohistochemistry.The percentage of T lympholeukocyte subset in peripheral blood of mouse was analyzed by flow cytometry.④To certificat the pathogenic mechanism of damp-heat syndrome of seasonal febrile disease induced by MHV—A59 through treating this disease with the decoction of clearing away heat and dampness:four-weeks-old mouse of BALB/c was divided into five groups randomly,control group,model group,A Decoction group(mouse of model group was treated by Decoction of Hao qin qing dan),B Decoction group(mouse of model group was treated by Coptidis Decoction for Detoxification),C Decoction group(mouse of model group was treated by Three-seeds Decoction).The quantity of MHV-A59 in mouse liver tissue was detected by FQ-PCR.The level of IL-4 and IFN-γin mouse serum was detected by ELISA.The expression of NF-κB in mouse liver tissue and AQP4 in gastric mucous membrane was analyzed by immunohistochemistry.The percentage of T lympholeukocyte subset in peripheral blood of mouse was analyzed by flow cytometry.3.Clinical research:The 30 patients of exogenous damp-heat syndrome were from fever caused by exogenous pathogens out-patient clinic of the first affiliated hospital of guangzhou university of Chinese medicine.There were 15 healthy volunteers as control group.The level of IL-4 and IFN-γof patients and healthy volunteer in serum were detected by ELISA.The patient was treated by Decoction of Hao qin qing dan for three days.then we detected the level of IL-4 and IFN-γ,again.Results:1.literature research:In current study,researchers have not used virus to duplicating model of damp-heat syndrome of seasonal febrile disease,and analyze the pathogenic mechanism of damp-heat syndrome of seasonal febrile disease in various model and multistrata.It was an important method to certificate the pathogenic mechanism of damp-heat syndrome of seasonal febrile disease through treating this disease with the decoction of clearing away heat and dampness.MHV-A59,as one of suitable succedaneum of human coronary virus,play a important role in duplicating model of damp-heat syndrome of seasonal febrile disease.2.experiment research:①The level of TG and TNF-αin serum of two model group obviously risen up compared to control group(p＜0.01).The mouse of two model group show up fever,languor,Diarrhea,weight lose,poor appetite etc.②The expression of NF-κB and AQP4 of model group and Bacillus coli group obviously risen up compared to control group(p＜0.05 or p＜0.01).The Th1/Th2(IFN-γ/IL-4)of model group and Bacillus coli group obviously risen up compared to control group(p＜0.05 or p＜0.01).The balance of CD4~+/CD8~+ of model group became obviously disequilibrium compared to control group (p＜0.01).③The quantity of MHV-A59 in mouse liver tissue of model group decreased compared to MHV-A59 group,but the difference was not significant(p＞0.05).The expression of NF-κB of model group and MHV-A59 group obviously risen up compared to control group(p＜0.01),but there were not significant different between damp-heat group and control group(p＞0.05),same as between model group and MHV-A59 group(p＞0.05). The expression of AQP4 of model group and damp-heat group obviously risen up compared to control group(p＜0.05),but there were not significant different between MHV-A59 group and control group(p＞0.05),same as between model group and damp-heat group(p＞0.05).The balance of Th1/Th2 and CD4~+/CD8~+ of model group became obviously disequilibrium compared to control group(p＜0.05)because of MHV-A59 infecting.④The quantity of MHV-A59 in mouse liver tissue of A decoction group decreased compared to model group,but the difference was not significant(p＞0.05).The expression of NF-κB of B decoction group obviously decreased compared to model group(p＜0.05).The expression AQP4 of C decoction group decreased compared to model group,but the difference was not significant(p＞0.05).The disequilibrium of Th1/Th2 was adjusted obviously compared to model group(p＜0.05).The disequilibrium of CD4~+/CD8~+ was adjusted,but the difference was not significant(p＞0.05)3.Clinical research:The level of IL-4 was not significant different(p＞0.05)between patients of exogenous damp-heat syndrome and healthy volunteer,but the level of IFN-γof patients obviously risen up compared to healthy volunteer(p＜0.01).The disequilibrium of Th1/Th2 was obvious compared to healthy volunteer(p＜0.01).The proportionality of Th1/Th2 of patients who treated by decoction of Hao qin qing dan obviously descend (p＜0.01).Conclusion:According to documents,the model of damp-heat syndrome of seasonal febrile disease could be duplicated perfectly by compound factors including richly fatty and sweet diet, humid heat environment and infecting by biology causative agent.Pathological changes character of animal model was determined by biology causative agent.Richly fatty and sweet diet and humid heat environment led to damp-heat.We could draw five conclusions through our study.Firstly,we duplicated a new mouse model of damp-heat syndrome of seasonal febrile disease by richly fatty and sweet diet,humid heat environment,MHV-A59. The model possessed characteristics of the illness and syndrome.Secondly,the expression of NF-κB in mouse liver tissue and AQP4 in gastric mucous membrane were one of the mutual pathogenic mechanism of damp-heat syndrome of seasonal febrile disease.The disequilibrium of Th1/Th2 in serum was another mutual pathogenic mechanism.But the disequilibrium of CD4~+/CD8~+ was only appeared in damp-heat syndrome of seasonal febrile disease induced by MHV-A59.Thirdly,MHV-A59 play a key role in inflammatory reaction and disorder of T cell immunity of damp-heat syndrome of seasonal febrile disease,and richly fatty and sweet diet and humid heat environment lead to water metabolism disorder.Quartly,the decoction of clearing away heat and dampness could treat damp-heat syndrome of seasonal febrile disease by affecting above-mentioned target,it was the evidence for the pathogenic mechanism referred in our study.Finally,according to clinical research,the disequilibrium of Th1/Th2 was one of the pathogenic mechanism of damp-heat syndrome of seasonal febrile disease,the decoction of Hao qin qing dan could treat this disease through adjusting disequilibrium.
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