基于清营汤探讨"透热转气"干预TLR-NF-κB信号通路影响脓毒症转归的效应机制

" Into the ying a diathermy gas transfer " is first put forward the theory of Ye Tianshi, " through heat transfer and gas " has important theoretical and practical significance for guiding the treatment of warm diseases. The representative prescription for ying Wu Jutong clear soup, which has both horns, Radix Scrophulariae, antelope horn of heat-clearing and detoxifying drugs, honeysuckle, forsythia, also leaves through heat transfer and gas drugs. " Through heat transfer and gas " contains the scientific connotation, is one of the key scientific issues in the development of traditional Chinese medicine. Based on " into the camp a diathermy to Qi " theory, to explore Qingying Decoction of diathermy gas transfer effect of the drug on septic rats. This project is to copy and improvement of septic rats as models of disease, with Qingying Decoction and different drugs, as a means of intervention of sepsis, to represent the different stages of the development of " TLR -NF-κB pathway " of the promoter of TNF- α, Toll like receptor ( TLR ), a negative regulator of beta -arrestin, regulator NF- κB, effector IL-6, reverse IL-10 cell effector factor as target. Discussion of Qingying Decoction and heating gas transfer of drugs on sepsis in " TLR -NF-κB " pathway of inflammatory mediators " waterfall cascade " the role of the target and control mechanism, in order to further reveal the TCM " through heat transfer and gas " contains the scientific connotation, " through heat transfer gas " put forward the theory of according to the treatment of sepsis, and provide the experimental basis for Qingying Decoction in the treatment of sepsis.

"入营犹可透热转气"是叶天士首先提出的理论，对于指导温热病的治疗具有重要理论和实践意义。其代表方剂为吴鞠通的清营汤，其中既有犀角、生地之清热解毒药物，也有银花、连翘、竹叶透热转气药物。现基于"入营犹可透热转气"理论，探讨清营汤中透热转气药物对大鼠脓毒症时的影响。本项目以复制并改良大鼠脓毒症为疾病模型，以清营汤及不同药物配伍，为干预手段，以代表脓毒症发展不同阶段"TLR -NF-κB通路"的启动子TNF-α、Toll 样受体(TLR)、负调节因子β-arrestin、调控子NF-κB、效应子IL-6、反向效应子IL-10细胞因子为目标。探讨清营汤及透热转气药物对脓毒症中"TLR -NF-κB"通路炎症介质的"瀑布样级联反应"的作用靶点及调控机理，以期进一步揭示中医药"透热转气"中蕴含的科学内涵，为"透热转气"治疗脓毒症提出了理论依据，为清营汤治疗脓毒症提供实验基础。

脓毒血症的发病机制非常复杂，TLR-NF-kB通路是炎症反应时机体产生免疫应答和炎症反应的关键信号系统。本研究基于清营汤配伍规律研究透热转气药对脓毒症有关指标的影响：分别从大鼠TNF-α、TLR4、β-arrestin、NF-kB p65、IL-6、IL-10指标，观察位于TLR-NF-kB上中下游水平信号效果。阐述基于“透热转气”理论的清营汤对脓毒症的“TLR -NF-κB通路”级联反应的作用靶点及调控机理。.此研究将150只雄性Wistar大鼠，随机分为5组：假手术组、脓毒症模型组、清营汤整方组、清热解毒拆方组和透热转气拆方组；每组再分为3 h、6 h和12 h三个亚组，每亚组10只。采用经典的盲肠结扎穿刺术（CLP）制备大鼠脓毒症模型。.在造模后3h各组大鼠血清TNF-α、IL-6和IL-10水平无差异；在造模后6 h和12 h，与假手术组比较,脓毒症组大鼠血清TNF-α、IL-6水平升高，IL-10水平下降，与脓毒症组比较，各治疗组大鼠血清TNF-α、IL-6水平降低，IL-10水平升高有统计学意义。于造模后6 h，各治疗组肺组织病理损伤、肝组织病理损伤有所改善，其中以清营汤组效果最为明显。与模型组比较，清营汤组大鼠肝组织TLR4、NF-kB蛋白表达降低，β-arrestin表达升高有显著性，但清热解毒组和透热转气组TLR4、NF-kB和β-arrestin蛋白表达较模型组差异无显著性。.清营汤能减轻脓毒症大鼠肺、肝病理损伤和中性粒细胞浸润，降低血清炎性细胞因子TNF-α、IL-6浓度，提高IL-10水平，这一作用与促进β-arrestin蛋白表达，抑制TLR -NF-kB信号通路有关。清热解毒组和透热转气组，无统计学差异，充分说明透热转气在清营汤中的作用非常重要，透热转气组是通过上游及下游水平对于脓毒血症进行干预的。