目的 探讨桃红四物汤（Taohong Siwu Decoction，THSWD）抗血栓形成的药理作用及机制。方法 通过ip卡拉胶溶液（10 mL/kg）建立小鼠血栓模型，将小鼠随机分为对照组、模型组、阳性药组和THSWD低、中、高剂量（7.74、15.48、30.96 g/kg）组，观察小鼠黑尾长度，检测血清和肝脏中相关因子；同时，进行凝血酶诱导血小板激活和脂多糖（lipopolysaccharide，LPS）诱导人脐静脉内皮细胞（human umbilical vein endothelial cell，HUVECs）损伤体外实验，检测血小板和内皮细胞中相关因子，观察THSWD对血小板和内皮细胞黏附作用的影响。结果 与模型组比较，THSWD各剂量组可显著减少小鼠体内的血栓并显著降低小鼠肝脏中高迁移率族蛋白B1（high mobility group box 1 protein，HMGB1）的水平（P＜0.01）和血清白细胞介素-1β（interleukin-1β，IL-1β）、IL-6、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）的水平（P＜0.01）；显著升高小鼠血清中组织型纤溶酶原激活剂（tissue plasminogen activator，TPA）的水平（P＜0.01）；此外，THSWD各剂量组还可显著降低肝脏中核因子-κB（nuclear factor-κB，NF-κB）和核苷酸结合寡聚化结构域样受体蛋白3（nucleotide-binding oligomerization domain-like receptor pyrin domain containing 3，NLRP3）的mRNA表达（P＜0.05、0.01）。体外实验发现，THSWD可升高血小板中超氧化物歧化酶（superoxide dismutase，SOD）的水平（P＜0.05）、抑制血小板凝块收缩、降低细胞外调节蛋白激酶1/2（extracellular-regulated kinase 1/2，ERK1/2）和HMGB1的蛋白表达（P＜0.05）、有降低蛋白激酶B（protein kinase B，Akt）蛋白表达的趋势；THSWD能显著降低LPS诱导的HUVECs损伤过程中活性氧（reactive oxygen species，ROS）的水平（P＜0.01）、抑制相关黏附因子血管细胞黏附分子-1（vascular cell adhesion molecule-1，VCAM-1）和细胞间黏附分子-1（intercellular cell adhesion molecule-1，ICAM-1）的表达（P＜0.05、0.01）、减少HUVECs与血小板的黏附（P＜0.05）。结论 THSWD可通过抑制炎症反应、促进纤维蛋白酶原激活剂生成、抑制氧化应激反应、抑制血小板激活、抑制内皮细胞损伤及与血小板的黏附发挥抗血栓作用。

Objective To explore the pharmacological effect and mechanism of antithrombosis in Taohong Siwu Decoction (桃红四物汤, THSWD). Methods The thrombus model was prepared by intraperitoneal injection of carrageenan. Mice were randomly divided into control group, model group, positive group and THSWD treatment groups (7.74, 15.48, 30.96 g/kg), observing the mice black-tail length, associated factors in the serum and liver were tested. Performing thrombin-induced platelet activation and lipopolysaccharide (LPS) induced injury of human umbilical vein endothelial cells (HUVECs) in vitro, using the benchmark sample of THSWD to administration, testing and analyzing related indicators, and observing the effect of THSWD on the adhesion of platelets and endothelial cells. Results Compared to the model group, each dose group of the THSWD can significantly reduce the thrombosis in mice (P < 0.01). High mobility group box 1 protein (HMGB1) in the liver and the levels of interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α) in the serum of the molded mice were significantly decreased (P < 0.01). THSWD significantly increased the level of tissue plasminogen activator (TPA) in serum (P < 0.01). In addition, THSWD can also significantly reduce nuclear faction-κB (NF-κB) and nucleotide-binding oligomerization domain-like receptor pyrin domain containing (NLRP3) mRNA expressions in liver (P < 0.05, 0.01). The results of in vitro experiments indicated that THSWD could increase the level of superoxide dismutase (SOD) in platelets (P < 0.05), suppress platelet clot contraction, reduce the expression of extracellular regulated kinase 1/2 (ERK1/2) and HMGB1 (P < 0.05), and exhibit a tendency to decrease protein kinase B (Akt) protein expression. In addition, THSWD can significantly reduce reactive oxygen species (ROS) levels during injury to HUVECs induced by LPS (P < 0.01), inhibit expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular cell adhesion molecule-1 (ICAM-1) (P < 0.05, 0.01), and reduce HUVECs adhesion to platelets (P < 0.05). Conclusion THSWD can inhibit inflammation, promote the production of fibrinogen activator, inhibit oxidative stress response, inhibit platelet activation, inhibit endothelial cell injury and adhesion to platelets, and play an antithrombotic role.

R285.5

四川省自然科学基金项目（2024NSFSC0696）