“大肠主津”理论指导下，柴芩承气汤缓解重症急性胰腺炎毛细血管渗漏的防漏堵渗机制研究

Capillary leak syndrome (CLS) is one of the major reasons for the occurrence of acute organ failure induced by severe acute pancreatitis (SAP), which is closely related to the activation of p38MAPK signaling pathway of capillary endothelial cells and still has no effective therapy in western medicine. The occurrence of Yangming Fushi of severe acute pancreatitis and the intestinal endotoxemia were reinforced each other. the theory of “the large intestine governs Jin” consided that the large intestine is the organ which regulates the distribution of body fluid. The therapy of Catharsis such as Chaiqingchengqi decoction (CQCQD) can Jun hot junction and diarrhea the dry air of Yangming large intestine, and then save its Jin. On the basis of the CQCQD could promote the recovery of gastrointestinal function and relieve pathological organ damage,for example, lung injury caused by acute necrotizing pancreatitis(ANP) confirmed by our previous studies, we hypothesized that the mitigation of organ damage of SAP by the CQCQD was related to “the large intestine governs Jin” inhibition of p38MAPK signaling pathway on capillary endothelial cells, relieving the CLS of SAP. So the mRNA and protein expression of p38MAPK signal pathway ,the apoptosis and intercellular structural changes of the pulmonary capillary endothelial cells were detected, to study the effection of CQCQD on the p38MAPK signal pathway of capillary endothelial cells. And then get the innovative discovery for the perfection of the theories of prevention and treatment of SAP with Traditional Chinese medicine.

毛细血管渗漏综合征是重症急性胰腺炎器官损伤的重要因素，其发生与内皮细胞p38丝裂原活化蛋白激酶信号通路被内毒素及炎性介子激活密切相关；重症急性胰腺炎发生阳明腑实证与肠源性内毒素血症互为因果；“大肠主津”理论认为：大肠调节机体水电解质平衡，柴芩承气汤等通里攻下治疗可峻下热结、泻大肠阳明之燥气而救其津；我们在前期研究证实柴芩承气汤可促进急性坏死性胰腺炎胃肠动力恢复，减少促炎因子释放，减轻器官病理损害的基础上，提出假说：柴芩承气汤减轻重症急性胰腺炎器官损害与“大肠主津”抑制毛细血管内皮细胞p38丝裂原活化蛋白激酶信号传导通路，缓解毛细血管渗漏有关。本项目检测急性坏死性胰腺炎大鼠肺毛细血管内皮细胞p38丝裂原活化蛋白激酶信号通路相关基因和蛋白表达及凋亡和细胞间结构的变化，研究柴芩承气汤对该病毛细血管内皮细胞p38丝裂原活化蛋白激酶信号通路的影响，为完善中药防治重症急性胰腺炎理论体系获得创新性发现。

毛细血管渗漏综合征（CLS）是重症急性胰腺炎发生器官衰竭的重要原因之一，与毛细血管内皮细胞p38MAPK信号通路激活密切相关。 “大肠主津”理论认为大肠是调节体液分布的脏器；重症急性胰腺炎病机为热结阳明大肠而伤津，通里攻下法可泻阳明之燥气而救其津液；在前期研究证实：柴芩承气汤可促进重症急性胰腺炎胃肠功能恢复，缓解SAP器官组织病理损害。本研究为验证柴芩承气汤缓解重症急性胰腺炎缓解毛细血管渗漏综合征与抑制毛细血管内皮细胞p38MAPK信号通路有关。直接以毛细血管内皮细胞丰富的肺组织为研究对象，以SB203580做阳性对照并联合运用基因芯片、KEEG信号富集分析和蛋白互作网络进一步发掘CQCQD治疗CLS的作用靶。本研究发现CQCQD可减轻ANP大鼠胰腺病理评分和肺病理评分、血清及肺腔灌洗液TNF-α和IL-1β浓度、MPO活性、肺组织EB渗出、肺TNF-α和IL-1β蛋白及mRNA水平、P-NFκB、P-IκB蛋白，并升高ANP大鼠的PaO2及肺脏组织中Tyr-tubulin、Ace-tubulin和P- stathmin1表达。同时，生物信息学分析结果发现CQCQD治疗ALI差异表达基因主要涉及炎症反应（Toll样受体信号通路）、细胞连接（紧密连接、粘附连接、细胞骨架蛋白调控）及凋亡等生物学过程及通路，而Fos、Pik3r1和Src是CQCQD治疗ALI毛细血管内皮细胞渗漏的3个潜在核心基因。为进一步验证该预测，人脐静脉内皮细胞体外实验使用PP1、LY294002或c-Fos siRNAs 分别抑制Src、Pik3r1和Fos，发现三者均可抑制LPS刺激增高的细胞旁路渗漏及炎症因子释放；而且CQCQD可模拟前述三者抑制LPS诱导的p-Src、p-p85和c-Fos升高，从而减轻细胞旁路渗漏及炎症因子释放。因此，本课题证实了CQCQD可减轻SAP相关ALI的CLS，可能与抑制ANP肺p38的活化、减少炎症因子释放、维持SAP肺内皮细胞微管稳定相关；并进一步拓展CQCQD治疗CLS的潜在作用靶点，为中医药治疗SAP 的理论体系提供分子生物学证据。