土鳖虫通过改善炎症抑制深静脉血栓形成的机制研究  

Eupolyphaga Sinensis Walker Inhibits Deep Vein Thrombosis via Anti-inflammation

作　　者：罗淑芳 黄博宁 何广铭 桂蜀华[4] 詹雅娴 张威鹏 林宝琴 Luo Shufang;Huang Boning;He Guangming;Gui Shuhua;Zhan Yaxian;Zhang Weipeng;Lin Baoqin(The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Censorship Center,Market Supervision Administration of Shenzhen Municipality,Shenzhen 518071,China;Guangdong Yifang Pharmaceutical Co.Ltd.,Foshan 528244,China;School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China)

机构地区：[1]广州中医药大学第一附属医院,广州510405 [2]深圳市市场监督管理局许可审查中心,深圳518071 [3]广东一方制药有限公司,佛山528244 [4]广州中医药大学中药学院,广州510006

出　　处：《世界科学技术-中医药现代化》2022年第12期4917-4926,共10页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：广州市科学技术局基础研究计划项目(202002030108):从SIRT1/NF-kB抑制炎性细胞浸润研究丹红化瘀口服液干预深静脉血栓形成的分子机制,负责人:林宝琴。

摘　　要：目的探讨土鳖虫抑制深静脉血栓形成(Deep Vein Thrombosis,DVT)的作用机制。方法将70只SD(Sprague Dawley,SD)大鼠随机分为正常组、假手术组、模型组、土鳖虫低、高剂量组(0.52、1.04 g生药·kg^(-1)·d^(-1))、肝素组(200 U·kg^(-1)·d^(-1))、氯吡格雷组(25 mg·kg^(-1)·d^(-1))。建立狭窄法诱导的DVT大鼠模型。肝素组尾静脉注射(1 mL·kg^(-1)·d^(-1))给药;其他组灌胃给药(10 mL·kg^(-1)·d^(-1)),连续7天。于第6天造模,第7天给药后1.5 h取材。精密称量血栓湿重;腹主动脉采血,检测活化部分凝血活酶时间(Activated partial thromboplastin time,APTT)、凝血酶原时间(Prothrombin time,PT)、二磷酸腺苷(Adenosine diphosphate,ADP)诱导的血小板最大聚集率、全血粘度和血常规;采用酶联免疫吸附测定法(Enzyme-Linked Immuno Sorbent Assay,ELISA)检测大鼠血清中白细胞介素-1β(Interleukin-1β,IL-1β)和肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)水平;应用苏木精-伊红(Hematoxylin-Eosin,HE)染色法观察血栓和静脉壁的病理学变化及炎性细胞浸润情况并对炎性细胞分类计算;采用免疫组化技术分析血栓和静脉壁的组织因子(Tissue factor,TF)和白细胞分化抗原68(Leukocyte differentiation antigen 68,CD68)的表达;通过免疫荧光法测定血栓和静脉壁Ly6g蛋白的表达水平。结果与模型组比较,土鳖虫剂量依赖性地降低血栓湿重。土鳖虫对ADP诱导的血小板最大聚集率、APTT、PT、全血粘度以及血常规的各项指标均没有明显的影响,但剂量依赖性地降低血栓中炎性细胞的数量,血栓和静脉壁中TF蛋白表达,以及血清中IL-1β和TNF-α水平。结论土鳖虫可干预DVT,其作用机制可能与其改善炎症微环境有关。Objective To investigate the anti-deep vein thrombosis(DVT)mechanism of Eupolyphaga sinensis Walker(ESW).Methods Seventy SD rats were randomly divided into normal group,sham group,model group,ESW low and high dose groups(0.52 and 1.04 g crude drug·kg^(-1)·d^(-1)),heparin group(200 U·kg^(-1)·d^(-1)),and clopidogrel group(25 mg·kg^(-1)·d^(-1)).DVT rat model was established by narrowing inferior vena cava.Rats in the heparin group was given heparin by tail vein injection(1 mL·kg^(-1)·d^(-1)),and other groups were administered orally(10 mL·kg^(-1)·d^(-1))for 7 consecutive days.On the 6thday,rats were modeled.On the 7thday,samples were taken 1.5 h after the final administration.The wet weight of the thrombus was accurately weighed.Then blood was collected from the abdominal aorta to detect platelet aggregation rate,activated partial thromboplastin time(APTT),prothrombin time(PT),whole blood viscosity and blood cells.ELISA method was used to detect the serum levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α).In addition,hematoxylin-eosin staining(HE)method was used to observe leukocyte numbers in thrombus and vein wall.Moreover,the expression of tissue factor(TF)and leukocyte differentiation antigen 68(CD68)in thrombus and vein wall were detected by immunohistochemical technique.Furthermore,the expression of Ly6g protein in thrombus and vein wall was detected by immunofluorescence method.Results Compared with the model group,ESW decreased the weight of thrombus in a dose-dependent manner.Although ESW had no significant effects on maximum rate of platelet aggregation induced by ADP,APTT,PT,whole blood viscosity and blood cells,it decreased the number of leukocytes in thrombus and vein wall dose-dependently.ESW significantly reduced the expression of TF protein in the thrombus and vein wall,and the serum levels of IL-1βand TNF-α.Conclusion ESW inhibits DVT,and anti-DVT mechanism may attribute to its antiinflammation activity.

关 键 词：土鳖虫 深静脉血栓 炎性细胞 炎症 

分 类 号：R285.5[医药卫生—中药学]