Damp-heat Syndrome Essence Based On Experimental Study Of Inflammatory Signaling Pathway

Objective:Damp-heat syndrome is the most common syndrome of febrile disease in Lingnan area which is mostly caused by virus infection.Therefore,the treatment of depleting damp-heat is effective.On the basis of former animal model of damp-heat syndrome,this study optimized the method of making the damp-heat model by creating the damp-heat environment.Meanwhile,the study elucidated the objective foundation of damp-heat syndrome of febrile disease and discussed the mechanism and effect target of treatment of depleting damp-heat.Furthermore,the study made contribution to the revealment of the substance of damp-heat syndrome,the development of Chinese herb of depleting damp-heat could elevate the level of infectious diseases prevention by Chinese medicine.Method:On the basis of former damp-heat syndrome animal model,our study applied the“fat and sweet diet + damp-heat environment + rhubarb intragastric administration + influenza virus infection”method to make the damp-heat syndrome model.Whether the model was successful or not was evaluated by weight,temperature,drink volume,phenotype,tongue nature,tongue coat and the lung tissue section.After the treatment of depleting damp-heat(Haoqin qingdan decoction),I observed the change of LBPmRNA、CD14mRNA、TLR4mRNA、TNF-α mRNA、与 NF-KBpmRNA in lung tissue to evaluate the effect of treatment of depleting damp-heat.At the same time,the study used cool acrid exterior-resolving method(Yinqiao San)as a comparison and ribavirin as control group.Results:1.Model construction(1)The mice in model group began to lose weight,drink less water and became fatigue.The somnolence rate was 82%.Other symptoms included filthy hair,glossless on sole of feet,thin sloppy stool,filthy periannal,orchidoptosis and other symptoms of damp-heat syndrome.At the same time,I observed the color of tongue.(2)The lung tissue of mice in model group showed the adherence,necrosis and defluxio of the bronchia epithelial cells.There was lymphoproliferation in the Inflammation area beside blood vessels and bronchia and the alveolus interstitium was thickening.(3)The tongue section showed filiform papilla increase compared to the normal group.And part of the filiform papilla was broadened with irregular shape.The tongue coat was thickened.(4)CD14、TLR4、TNF-a in the mice of model group was increased which was the objective foundation of damp-heat syndrome.1.Treatment study(1)The weight of mice in treatment group was higher than mice in model group,especially the Haoqin qingdan decoction group(P<0.01).The temperature in Yinqiao San group was the lowest.After the virus implantation and the treatment,the drink amount decreased while the drink amount in the normal group was stable.(2)The thickness of tongue coat in model group and other groups were obviously different(p<0.01).Tongue coat in ribavirin and Haoqin qingdan decoction group was thinner while the Yinqiao San group was not different compared with model group.(3)Among three groups,inflammatory cells in ribavirin group was abreacted while the epithelial cells has repaired and the structures were recovered in Yinqiao San group with alveolar space narrowing and assembling of inflammatory cells which proved little effect.(4)Among three groups,the tongue coat was thinner in ribavirin group and Haoqin qingdan decoction group than model group indicating good effect while the tongue coat was thicker in Yinqiao San group indication effect unsatisfactory.(5)There was no difference of LBP gene in model mice or normal mice.LBP gene was decreased in Haoqin Qingdan decoction and Yinqiao San group(P<0.01).In vibavirin group,there was a increasing tendency but not statistically significant.(6)CD 14 gene in model mice was higher than normal mice which might be related with the disease.After the treatment of Haoqin Qingdan decoction and Yinqiao San,CD14 decreased(P>0.05)while there was a obvious decrease of CD14 in ribavirin group(P<0.01).(7)There was a increasing tendency of TLR4 gene in mice in model group and decreased after treatment(three treatment group),especially in Haoqin Qindan decoction group(P>0.05).(8)There was no obvious difference of NF-κB gene in model mice or normal mice.NF-kb in Haoqin Qingdan decoction group and Yinqiao San group was obviously decreased(P<0.05 and P<0.01)while no difference in ribavirin group.(9)TNF-a was increase in model group while the difference of TNFa between three treatment groups was not obvious.Conclusion:1.During the construction of the animal model of damp-heat syndrome,the comprehensive method of "fat and sweet diet + damp-heat environment + rhubarb intragastric administration + influenza virus infection" was successful.The predominant cause of the pathological change of mice was the virus while other factors influenced the time,severity and recovery.2.Our study demonstrated that somnolence,sloppy stool,hair lofty,tongue color and tongue coat could be used as the index of success of the models.And the increase of TNF-a、TLR4、CD14 expression could be the common target of virus infection and damp-heat and the reflection of the substance of damp-heat syndrome in febrile disease.3.Factors related to the treatment of depleting damp-heat(1)Reduce the expression of CD 14,TLR4 and LBP and prevent LPS from working so that the cell transition was blocked and the treatment worked.(2)Suppress the expression of NF-κB and the reaction before inflammation.4.Compared with cool acrid exterior-resolving method,method of depleting damp-heat had advantage of reducing the expression of TLR4 while no obvious prevalence in other targets.Keywords:febrile disease,animal model,method of depleting damp-heat,Haoqing Qingdan decoction,intervention,inflammatory factors.