登革热证候传变规律及相应治法的生物学基础研究

Dengue fever is a life-threaten disease in human being. However, there are still not effective drugs and vaccines. Although traditional Chinese medicine (TCM) shows obvious advantages in the treatment of dengue fever, the syndrome transmission principles of dengue fever and the selection evidences of corresponding therapies have not yet been fully elucidated. Based on previous work, we think that the syndrome transmission of dengue fever is reflected as the variety of clinical manifestation, and dependent on the immune response of virus and body. The biological basis is the interaction of viruses, antibodies, complements, inflammatory factors and the body, which can be intervened by the therapy prescription (e.g. detoxication formulas). Our previous work had collected dengue fever patients and established dengue fever, DENV-ADE mouse and cell models. Under the guide of theory of epidemic febrile diseases, the project aims to explore the syndrome transmission principles of dengue fever and corresponding therapies by pathogenic biology, infection immunology, molecular biology and bioinformatics methods. The research will clarify the crosstalk of viruses, antibodies, complements, inflammatory factors and the body in the pathological process of dengue fever. It will also elucidate the principles of corresponding therapies to different syndromes and the anti-dengue fever mechanisms of representative formulas, which will support a rationale for the prevention and treatment of dengue fever by TCM.

登革热发病机制尚未完全阐明，治疗药物及疫苗研发仍举步维艰。中医药治疗登革热具有优势与特色，但有关登革热证候传变规律并不明确，相应治法方药的选择依据尚欠充分。基于课题组先期临床与基础研究进展，我们认为：登革热证候传变体现为临床表现的多样性，取决于病毒与机体免疫应答，其生物学基础为病毒、抗体、补体、细胞因子及机体等因素相互影响、消长转化，以解毒法为代表的治法方药可针对上述因素发挥多途径干预作用。本项目以登革热患者及先期建立的登革热小鼠模型和DENV-ADE重症小鼠模型、细胞模型为研究对象，采用病原生物学、感染免疫学、分子生物学、生物信息学等方法，开展登革热证候传变规律及相应治法的生物学基础研究，明确病毒、抗体、补体、细胞因子、机体在登革热特别是重症登革热发生发展中的相互关系，揭示登革热证候传变规律，阐明不同阶段、不同证候相应治法的治疗原理和代表方药的作用机制，为登革热的中医药防治提供科学依据。

登革热发病机制尚未完全阐明，治疗药物及疫苗研发仍举步维艰。中医药治疗登革热具有优势与特色，但有关登革热证候传变规律并不明确，相应治法方药的选择依据尚欠充分。因此，本项目开展了登革热证候传变规律及相应治法的生物学基础研究。研究结果表明，登革热证型以卫气同病、瘀毒交结居多。邪毒致瘀、毒瘀交结为核心病机，具有毒、热、瘀、虚、兼夹湿邪证候传变规律。发热期湿热郁卫、卫气同病证候，采用体现清暑化湿、解毒透邪治法的柴石解毒方；极期毒瘀交结，扰营动血证候，采用体现解毒化瘀，清营凉血治法的解毒化瘀方、清瘟败毒饮及相关中成药；恢复期余邪未尽，气阴两伤证候，采用体现益气复脉，养阴生津治法的生脉饮等治疗。在对登革热证候传变的生物学基础研究中发现，TNF-α、IL-6、IFN-α2、IP-10和sVCAM-1有重症预警价值；登革病毒主要激活先天免疫应答、趋化因子相关信号通路及白细胞、单核细胞趋化作用，并且免疫应答强度与病情轻重及证候表现相关。药物作用机制研究发现，发热期所用的柴石解毒方可改善模型动物症状，降低病毒载量、炎症因子水平，减轻组织病理损伤。极期所用的清瘟败毒饮抑制NF-κB和STAT3通路活化，缓解过度炎症、减轻病理损害；凉膈散可抑制细胞膜Cav1表达阻止HSP70质膜转移，阻止E和HSP70结合，抑制DENV吸附进入，还可抑制炎症，改善血管渗漏、血小板降低、器官组织损伤；痰热清注射液抗DENV-2脑炎作用与抑制NLRP3激活，减轻炎性反应，减少IL-1β释放，保护神经元有关；恢复期所用的生脉饮能改善模型动物症状，降低病毒滴度，抑制炎症因子水平，减轻组织病理损伤，其抗炎作用与抑制NF-κB和STAT3通路活化有关。此外，本项目还首次建立了DENV-3感染糖尿病小鼠模型，改善了登革动物模型匮乏的现状，具有明显的创新性。综上，本研究揭示登革热证候传变规律，阐明不同阶段、不同证候相应治法的治疗原理和代表方药的作用机制，为登革热的中医药防治提供科学依据。