Establishment Of Mouse Model Serving For Febrile Disease Damp-Heat Syndrome In LingNan Region And Study On Essence Of FDDHS In The Intestines

PurposeThe formation of Febrile Disease Damp-Heat Syndrome(FDDHS)is based on constitution of Damp-Heat Syndrome,and subsequently suffer from exogenous pathogen.South of the Five Ridges refer to Guangdong,Guangxi,Hainan and other regions.Not only the topography is similar in this regions,but also eating habits is similar,people like eat high protein foods such as seafood,chicken,they also like eat dessert.Damp-heat syndrome is a common type of constitution in south of the Five Ridges,pathogenesis of damp-heat syndrome include deficiency of spleen and stomach,diet of high protein relate to deficiency of spleen and stomach.This study intends to establish mouse model of FDDHS,this aim is to explore composition of intestinal flora,Inflammatory status and the change of aquaporins in the Febrile Disease Damp-Heat Syndrome.We choose Yinchenhao decoction to treat disease,and explore the mechanism of Yinchenhao decoction.MethodsOur study establish Two kinds of models of Febrile Disease Damp-Heat Syndrome in LingNan Region,pathogenesis of Two kinds of models are different.We utilized LingNan Region-featured diet easily induces deficiency of the spleen and accumulation of endogenous dampness,we utilized LPS to simulate factor of Pandora gas,which couples with the environment of high temperature and high humidity to produce FDDHS.We estabilsh two kinds of models We utilized them to observe the tongue and the colon tissues by HE staining,to detect content of LPS,TNF-a in serum and colon by ELISA,to detect the characteristic of intestinal flora by PCR,to detect levels of TLR4,CD14,AQP3 and AQP8 in colon tissue by RT-PCR in the FDDHS.We choose Yinchenhao decoction to treat disease,and explore the mechanism of Yinchenhao decoction via levels of TNF-?,TLR4,CD14,AQP3,AQP8.ResultsTwo kinds of mouse model also showed identically clinical symptoms of FDDHS,content of TNF-? were markedly boosted in serum and in colon tissue in mice of A and B model group(P<0.05),this means that the model is successfully establish.At the beginning of model about 6 hours,content of LPS in serum in mice of A model group was less than content of LPS in serum in mice of LPS group(P>0.05),content of LPS in serum in mice of B model group was less than content of LPS in serum in mice of LPS group(P<0.05).It speculate that bacterial translocation occurred in mice of A and B model group,LPS come from intestinal bacteria caused LPS tolerance,LPS tolerance would showed that content of LPS were decreased in mice of A and B model group.Compared with control group,expressive levels of TLR4mRNA?CD14 mRNA were increased in colon tissue in mice of A and B model group,expressive levels of AQP3 mRNA were decreased in colon tissue in mice of A and B model group,expressive levels of AQP8 mRNA were increased in colon tissue in mice of A and B model group,it speculate that activation of TLR4 pathway and dysfunction of aquaporins occurred in the Febrile Disease Damp-Heat Syndrome.E.coli,Enterococcus and Clostridium were over-growth in two kinds of models,while two probiotics,Bifidobacterium and Lactobacillus genus,were up-regulated in mice of A model group but down-regulated in mice of B model,it showed that Dysbiosis is a pivotal event during the onset of FDDHS.We use Yinchenhao decoction to treat FDDHS for 7 days,clinical symptoms of FDDHS improved in mice of A and B model group,Yinchenhao decoction can alleviate FDDHS.Our studys showed that content of TNF-? in serum in mice of A and B Yinchenhao decoction group was less than content of TNF-? in serum in mice of A and B saline group,it showed that Yinchenhao decoction can inbibite inflammatory.Compared with A and B saline group,expressive levels of TLR4mRNA?CD14 mRNA were decreased in colon tissue in mice of A and B Yinchenhao decoction group,Yinchenhao decoction could decrease expressive levels of TLR4mRNA?CD14 mRNA for anti-inflammatory.Yinchenhao decoction treat for 7 days,water quantity and diet were up-growth,moisture content of faeces were down-growth.Compared with A and B saline group,expressive levels ofAQP3mRNA were decreased in colon tissue in mice of A and B Yinchenhao decoction group,expressive levels ofAQP8 mRNA were increased in colon tissue in mice of A and B Yinchenhao decoction group,it showed that Yinchenhao decoction can regulate levels of AQP3,AQP8 in colon tissue.The A and B saline group respectively derived from A and B model group for 7 days,Compared with A and B model group,expressive levels of TLR4mRNA?CD 14 mRNA were increased in colon tissue in mice of A and B saline group,it showed that degree of activation of TLR4 pathway was aggravate as time goes on.Compared with A and B model group,expressive levels of AQP3mRNA were increased in colon tissue in mice of A and B saline group,levels of AQP8mRNA were increased in colon tissue in mice of A saline group,levels of AQP8mRNA were decreased in colon tissue in mice of B saline group,it showed that pathological changes about stagnation of fluid-dampness relate to expressive levels of AQP3,AQP8.ConclusionTwo kinds of mouse model in FDDHS in South of the Five Ridges is successfully establish,mice of Two kinds of model with systemic inflammatory response show that similar clinical symptoms of FDDHS.This is to express imbalance of composition of intestinal flora in Febrile Disease Damp-Heat Syndrome,,two kinds of models show that levels of pathogenic bacteria is overgrowth,such as Escherichia,enterococcus,Clostridium,Levels of Probiotics is affected by the pathogenic mechanism of FDDHS.Essence of FDDHS include activation of TLR4 pathway and disorder of levels of AQP3,AQP8.FDDHS can treat by Yinchenhao Decoction,mechanism of Yinchenhao Decoction is to inhibit activation of TLR4 pathway for anti-proinflammatory and regulate levels of AQP3,AQP8 for draining water.