Establishment Of Dengue Animal Model With Damp-heat Syndrome And The Study About Mechanism Of Action On Clearing Heat And Eliminating Dampness

Backgroud:Dengue fever popular at least had more than 200 years until now. The popular scale and frequency of DF have expanded endlessly since 20 cen-turies. DF once vanished more than 30 years in our country, Foshan reported Dengue fever epidemic situation for the first time in 1978, and then repeated in the popular southeast coast citys frequently. Among them, Guangdong Province is the hardest hit by dengue fever, and 63% events of the province occurred in Guangzhou. With the urbanization of Guangzhou, DF has a rising trend, how to effective control will affect it's economy and image directly. Effective prevention and controltion of DF, becomes a public health issue to be resolved hastily.There are no special effective methods for DF's treatment current-ly, mainly about symptomatic and supportive therapy. Treatments of TCM on DF has a unique effect, how to further study Chinese medicine, a suitable animal model will play a key pole.Objective:According to literature review and retrospective clinical Case Study, we hope that we will understand the profile of Chinese medicine treatment of dengue fever, as well as the weak link in previous models, and then to seek enlightenments on the study; of different rodents, different methods and different heat exposure attacking multiple factors such as drug models modeling of the effects in order to find an ideal animal model for dengue Damp; On this basis, verify the effect of heat treatment of dampness and the function of target, of damp and water channel protein.Methods:1,Literature:to finish a Chinese medicine clinic on the awareness and profile of dengue fever and dengue fever murine model of progress by literature reviews and analysis the damp-heat characteristic of dengue in Lingnan, which will be focuse on it's inspiring role in this experiment and clinical research. 2,Experimental Studies:there are two parts which were the model building and evaluation and the intervention effects of clearing heat and dampness.①,To establish Damp-heat model of dengue fever:The 4 weeks old BALB /c and C57BL/6 mice were divided into short-term heat acclimation and heat exposure, while divided into single dengue virus 2 infections (referred to as DF2) and ADE effects (the ADE) model of two different virus challenge respectively;So, in accordance with the principles of permutations and combinations, there were 8 model. Finally, there were 14 group adding the blank control group, DF2 control group, damp-heat group included 14 group. Then through the observation of mice body temperature, symptoms, signs, and platelet (PLT), liver function (ALT, AST), blood lipids (TG, CHOL) and liver pathology, to evaluate the results of model establishment and filter out the best damp-heat syndrome of dengue fever in animal;②,To compare and investigate the mechanism of the intervention effects of different drugs on mice of Damp-heat model of dengue fever: Mice selected by the first experiment were divided into 5 groups randomly: normal group, model group, Artemisia Scutellaria Decoction treatment group, treatment group Qingwenbaiduyin drink, Ribavirin treatment group, saline treated group. Then to observe body temperature, symptoms, signs, platelet (PLT), liver function A (LT, AST), blood lipids (TG, CHOL) and liver pathology, and to detect brain aquaporin 4 (AQP4), renal water channel protein 2 (AQP2), stomach aquaporin 4 (AQP4). Finally, to compare the therapeutic effects of different drug treatments and explore their therapeutic mechanism.3,Clinical Research:Review of 451 cases in-hospital in Guangzhou 8th People's Hospital in 2006; All cases were divided into damp-heat group and summer-dry group in accordance with certain standards, then to compare the age distribution, average age, gender, body temperature peak, the incidence of rash and impaired liver function (ALT, AST increase), plate-let levels, etc. Further to divided those 451 cases into two groups:group with combinative treatment and group with western medicine and compare the mean fever clearance time, rashes subside time, normalization of platelet time, the average duration of illness and ALT, AST levels, to determine its efficacy.Results:1,Literature Review:①The damp-heat factors of seasonal febrile disease in Lingnan had a certain effect on dengue fever.②Chinese medicine threpy in dengue fever had more effective than western medicine.③Animal model of dengue fever wasn't ideal. There were many reasons possibly, but it's likely to be the most important reason to have no breakthrough in thinking;④The combination of disease and syndrome maybe have a promising future. 2,Experimental Research:①,General Performance:After a virus attack, mice indicated fatigue condition, lower temperament, less movement, tired lying, slow respond to stimuli, hair standing on flow, less shiny, less diet and water intake, more smelly urine and soft feces. Into the warehouse, mice manifested irritability, less activities, less diet and respond to stimuli, more water intake and tired lying. After put into the warehouse several times. The difference of BALB/c and C57BL/6 mice wasn't significant. Haoqin-qingdan decoction had paid a certain role in improving the symptoms of model group.②,Body Temperature Test:After the virus attacks, the temperatures of mice slowed down, and then gradually returned to normal; short-term heat ex-posure could cause fever increasing, but became minute after heat ac-climation. The differences among the groups are significant. Both Hao-qinqingdan decoction and Qingwenbaiduyin decoction had the effects of decreasing body temperature.③,Detection of the main indexes:After virus attacks, PLT slowed down to a certain extent similarly. ALT and AST increased to varying degrees, especially AST;TG, CHOL increased in mice given high fat diet fed. But liver function and blood lipids had no significant increase in heat ac-climation group. So Haoqin decoction could decrease ALT, AST and TG, CHOL, while Qingwenbaidu decoction could upgrade PLT.④,After established modle with complex method, the expression of AQP4 in brain tissue and AQP2 in gastric mucosa increased, but renal AQP2 re- duced. Haoqin decoction could reverse those expressions to the nomal levels, and the differences were statistically significant, compared with other groups. Through correlation analysis three AQP above, the in-dependent variables of renal AQP2 had a significant correlation with the dependent variable of gastric AQP4 and The independent variables of renal AQP2 had no correlation with the dependent variable of gastric AQP4.⑤,Liver pathology:There are varying pathological changes in specimens of each groups. Liver cells had edema from mild to sever. Individual cells had eosinophilic changes;Liver cells had punctate, small focal necrosis, and necrosis area was infiltrate by inflammatory cells; There was no steatosis and fibrosis. Microscopy shows slight swelling of liver cells in group with damp-heat intervention; The state of liver cells had im-proved after the treatment of Haoqinqingdan decoction.3,Clinical Studies:①,Patients with dry-summer plague and damp-heat plague had dif-ferent characteristics in age distributionh, but no dif- ference in age, sex, concurrent liver damage. Summer-dry group were signi-ficantly higher than damp-heat group in the event of epidemic fever, rash and platelet decrease; ②,The integrated therapy group were less than the western medicine group in the average number of days fever, rash subsided days, the average duration of hospital stay, and summer-dry group were less than damp-heat group in those indexes;③,In platelet recovery time, the difference between the group with damp-heat plague and control group wasn't significant, but the group with summer-dry was less than the group with western therapy and control group and the difference was significant;④,After 7 days or 7-14 days treatment with integrative medicine, there was no difference between two groups with integrative medicine and the group with western medicine in liver function.Conclusion:1,Experimental Studies Conclusion:According with symptoms, fever, liver damage and blood lipid, the model group of BALB/c+DV2+thermal indicated damp-heat characterristic of dengue fever, so it's the best method. Haoqin decoction had therapeutic effects on mice with damp-heat syndrome of dengue fever, while have improved symptoms, decreased blood lipids and liver pathological damage, and regulated the water channel protein adjustment disorders;2,Clinical Research Findings:the dry-summer plague had more severe fever, skin rash and lower platelet than the damp-heat plague. Integrated Treatment had more significant effects than western medicine treatment in improvement those symptoms, in which the summer-dry group was better than damp-heat group, but the difference of improvement liver function wasn't significant.