A Study Of "Relieving And Eliminating Therapy Of Pyretic Stagnation" For Febrile Disease

Objective "Relieving and eliminating therapy of pyretic stagnation " for febrile disease is a kind of treatment combined "relieving"and "eliminating", based on the pathogenesis of "pyretic stagnation". This combinated treatment contrapose the pathological change of intermingling between warm pathogen and depression of Qi, aimed at dispelling exogenous pathogen. Since it has little record in traditional chinese medicine literature, the particular therapy of febrile disease needs objectify and normalization. Thus, it is essential to strengthen the investigation in this field through the combination oftraditional research methods and modern science and technology. According to this core thought, our study carry out from 3 parts: theoretical research, data mining and experimental study.Method Theoretical research:Systematically arranged the concept of febrile disease "relieving and eliminating therapy of pyretic stagnation", basis of this treatment, indications, concrete applications in febrile disease, in the understanding of this method by ancient physicians as well as the application of this treatment in pulmonary infectious diseases and so on. The second part used inverse mining approach. Based on 120 medical works from late Ming dynasty to the republic of China(1642-1949), recorded in "Encyclopedia Of Febrile Disease"written by Cao Hongxin, application informations of febrile disease "relieving and eliminating therapy of pyretic stagnation" were obtained, created into database, mined and analysed by data mining system Medcase V3.2. Through data mining of the symptom and prescription et al. for this particular therapy in febrile disease, formula-syndrome relationship of "relieving and eliminating therapy of pyretic stagnation" was analysed and investigated, providing objective basis and methodology reference in clinical application.The third part was experimental study. SJS of different concentration (0、2、 10μg/mL) was used to induce rat pulmonary alveolar macrophage cell NR8383. The expression level of TLR4 was measured by QRT-PCR and western blot separately. The mRNA level of TNF-a, A20, IL-6, IL-1β was measured by QRT-PCR, and the production of TNF-α, IL-6, IL-1β in the supernatant of NR8383 was determined with ELISA. In order to study the role of SJP in LPS-stimulated inflammation, we determined whether SJP would inhibit key pro-inflammatory mediators and cytokines including nitric oxide (NO) by Nitrite Assay, prostaglandin E2 (PGE2), and inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and tumor necrosis factor-a (TNF-a), IL-lbeta and IL-6 in lipopolysaccharide (LPS)-stimulated BR8383 cells via Elisa and Q-PCR, respectly. Then, the important pro-inflammatory transcription factors, NF-κB p65 and inhibitory kappaB-alpha (IκB-α), and mitogen-activated protein kinases (MAPKs), consist of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 were analyzed by Western blotting.Result1 Theoretical researchOur study find that "relieving and eliminating therapy of pyretic stagnation " for febrile disease combined "relieving"and "eliminating", is based on the pathogenesis of "pyretic stagnation". Main emphasis of "relieving" is to remove the pathogens from intensified to weak. Meanwhile, "eliminating" emphasised on dispersion and evacuation of pathogens, clearing the obstacles for outside the pathogens. Cooperation of the two effects will separate pyrexia and stagnation effectively and pathogenic factors are eliminated. Four stages of febrile disease, "weifen", "qifen", "yingfen" and "xuefen", could use it compatibly. There is a little record in traditional chinese medicine literature about the particular therapy of febrile disease without objectify and normalization. Modern scholars has found that "pyretic stagnation" related symptoms for have three features of high incidence, high fatality rate and strong resistance through retrospective review and analysis. Other scholars have applied the treatment of "relieving and eliminating therapy of pyretic stagnation " into pulmonary infectious diseases, and achieved satisfactory clinical curative effect. Cao Hongxin et al. retrospectively analyzed the curative effect of SARS treatments on patients of Beijing, Guangzhou, Tianjin and other places in 2003, the results showed that SJS ranked first in the application of comprehensive prescription compatibility. The above content, hot depressive warm disease through drainage method has a certain theoretical basis and the value of modern clinical research. As shown above, febrile disease "relieving and eliminating therapy of pyretic stagnation " has a certain theoretical foundation and great value of modern clinical research.2 Data mining21 Medical Books,181 medical cases,147 involving symptoms,15 types of pulse,51 tongue presentations,186 kinds of drugs,30 rules from symptom associations,30 rules from associations in tongue presentations,30 rules of pulse associations,94 rules from medication associations were retrieve and obtained relevant to febrile disease "relieving and eliminating therapy of pyretic stagnation ". Through data analysis of 181 cases of febrile disease "relieving and eliminating therapy of pyretic stagnation " that meet certain criteria, syndrome differentiation evidence of clinical application are as follows:common symptoms:contradictory febrile(such as febrile, conjunctival congestion, scanty dark urine, but pale complexion, no sweat or perspiration of forehead, cool limbs), consciousness disorder(dysphoria, coma and delirium), depressed vital energy(chest congestion, short breath). Visible symptoms:aversion to cold, rigor, headache, lips desiccation fissure, cough, expectoration, swollen sore throat, inappetence, emesis, nausea, oneirodynia, insomnia, constipation, loose stool, lassitude, asthenia, exanthesis. Tongue presentations:red tongue, yellow and greasy fur. Research about compatibility of drugs mode:①When given medications, the proportion of drug species and frequency for "eliminating drugs" are both higher than the other, "relieving drugs" have one thing of nature and flavour in common, pungent flavour or fragrance②Prescriptions of febrile disease "relieving and eliminating therapy of pyretic stagnation " are not the result of "eliminating drugs" add "relieving drugs", but other drugs exit. ③Shared common drugs:fermented soybean and capejasmine, cornu rhinoceri asiatici and fructus forsythiae, fructus forsythiae and Rehmannia Root, fructus forsythiae and figwort root.3 Experimental study.①Through the establishment of HPLC method for simultaneous determination in SJS powder of rhein, emodin, rhubarb phenol, we evaluated and confirmed the stability and reliability of the herb in prescription, providing references of quality control for the further experiments.②LPS induced TLR4 overpression in both mRNA and protein levels. Moreover, NF-κB signaling pathway and downstream target cytokines were increased with dose-dependent. SJP (5 μg/ml) significantly downregulated the protein expression levels of iNOS and COX-2 in LPS-stimulated cells, as well as the downstream products, containing NO, PGE2 and TNF-a, IL-1beta and IL-6. Moreover, SJP also prevent LPS-induced NF-κB p65 activation via phosphorylation of IκB-α and subsequent translocation of the NF-κB p65 subunit to the nucleus.