[关键词]
[摘要]
目的 考察桃核承气汤对脓毒症凝血功能障碍的作用及机制。方法 将 60 只 KM 小鼠按体质量随机分 为正常组,模型组,桃核承气汤低、高剂量组,阳性药肝素钠注射液组和血必净注射液组。除正常组外,其余 各组均腹腔注射角叉菜胶(Car)和脂多糖(LPS)制备小鼠脓毒症模型。测量小鼠尾部血栓相对长度和耳廓毛细血 管相对瘀血面积;试剂盒检测活化部分凝血活酶时间(APTT)和凝血酶原时间(PT);ELISA 法检测小鼠血清中 组织因子(TF)、C 反应蛋白(CRP)和白细胞介素 6(IL-6)水平;苏木素-伊红(HE)染色观察小鼠尾巴、肝脏和 肺组织病理变化;免疫组化法检测小鼠尾部血栓中血管性血友病因子(vWF)和肺组织中血管内皮钙黏蛋白 (VE-Cadherin)蛋白表达;流式多因子检测小鼠血浆中单核细胞趋化蛋白 1(MCP-1)、血管细胞黏附分子 1 (VCAM-1)、多配体蛋白聚糖 1(Syndecan-1)水平。脂多糖刺激 EA.hy926 细胞制备血管内皮损伤模型,RT-PCR 法检测 EA.hy926 细胞白细胞介素 1β(IL-1β)、IL-6 和 TF mRNA 水平。结果 与模型组比较,桃核承气汤明 显减少造模后 6 h 小鼠的耳廓相对瘀血面积(P<0.01),缩短尾部血栓相对长度(P<0.01),缩短造模后 12 h APTT 及 PT(P<0.01),降低血清 TF 水平(P<0.01)和 IL-6 水平(P<0.05),降低血浆 MCP-1、VCAM-1 和 Syndecan-1 水平(P<0.05,P<0.01),降低外周血白细胞计数(P<0.01),减少尾静脉管腔血栓面积(P<0.01), 减少肝、肺组织血管内瘀血(P<0.01),降低 vWF 和 VE-Cadherin 蛋白表达(P<0.05,P<0.01)。桃核承气汤 含药血清可明显降低脂多糖诱导 EA.hy926 细胞的 IL-6、IL-1β、TF mRNA 水平(P<0.05,P<0.01)。结论 桃核承气汤通过保护血管内皮预防脓毒症凝血功能障碍。
[Key word]
[Abstract]
Objective To investigate the effect and mechanism of Taohe Chengqi Decoction (THCQD) on coagulation dysfunction in sepsis. Methods Sixty KM mice were randomly divided into six groups based on body mass:normal group,model group,low-dose THCQD group,high-dose THCQD group,positive control Heparin Sodium Injection group,and Xuebijing Injection group. Except for the normal group,sepsis models were established in the other groups by intraperitoneal injection of carrageenan (Car) and lipopolysaccharide (LPS). The relative length of tail thrombosis and the relative congestion area of auricular capillaries were measured. Activated partial thromboplastin time (APTT) and prothrombin time (PT) were detected using reagent kits. Serum levels of tissue factor (TF),C-reactive protein (CRP),and interleukin-6(IL-6) were measured by ELISA. Pathological changes in the tail,liver,and lung tissues were observed using hematoxylin-eosin (HE) staining. Immunohistochemistry was used to detect the protein expression of von Willebrand factor (vWF) in tail thrombi and vascular endothelial cadherin (VE-Cadherin) in lung tissues. Levels of monocyte chemoattractant protein-1(MCP-1),vascular cell adhesion molecule-1(VCAM-1),and Syndecan-1 in plasma were measured using flow cytometry. An endothelial injury model was established by stimulating EA.hy926 cells with LPS,and RT-PCR was used to detect mRNA levels of interleukin-1β(IL-1β),IL-6,and TF in EA. hy926 cells. Results Compared with the model group,THCQD significantly reduced the auricular congestion area (P<0.01) and shortened the relative length of tail thrombosis (P<0.01) at 6 hours post-modeling,shortened APTT and PT (P< 0.01) at 12 hours post-modeling,decreased serum TF (P<0.01) and IL-6 levels (P<0.05),reduced plasma levels of MCP-1,VCAM-1,and Syndecan-1(P<0.05,P<0.01),lowered peripheral white blood cell counts (P<0.01), decreased thrombus area in tail veins (P<0.01),reduced intravascular congestion in liver and lung tissues (P<0.01), and downregulated the protein expression of vWF and VE-Cadherin (P<0.05,P<0.01). THCQD-containing serum significantly reduced LPS-induced mRNA levels of IL-6,IL-1β,and TF in EA.hy926 cells (P<0.05,P<0.01). Conclusion Taohe Chengqi Decoction prevents coagulation dysfunction in sepsis by protecting vascular endothelium.
[中图分类号]
R285.5
[基金项目]
国家自然科学基金委青年基金项目(82404817);国家自然科学基金青年项目(82104504);广州市科技局基础与应用基础项目之广州市 重点实验室建设项目(2024A03J0360);国家中医药传承创新中心科研专项(2023ZD01)。