食品科学 ›› 2022, Vol. 43 ›› Issue (15): 134-140.doi: 10.7506/spkx1002-6630-20210818-242

• 营养卫生 • 上一篇    

榆干离褶伞溶栓酶对动脉血栓大鼠血管内膜的保护作用

崔日花,李钰,苏新,卫莹,陈佳芮,沈明花   

  1. (1.延边大学附属医院,吉林 延吉 133000;2.延边大学医学院,吉林 延吉 133002)
  • 发布日期:2022-08-30
  • 基金资助:
    国家自然科学基金地区科学基金项目(81760031)

Protective Effect of Lyophyllum ulmarium Fibrinolytic Enzyme on Vascular Intima in Rats with Arterial Thrombosis

CUI Rihua, LI Yu, SU Xin, WEI Ying, CHEN Jiarui, SHEN Minghua   

  1. (1. Yanbian University Hospital, Yanji 133000, China; 2. College of Medicine, Yanbian University, Yanji 133002, China)
  • Published:2022-08-30

摘要: 目的:探讨榆干离褶伞溶栓酶(Lyophyllum ulmarium fibrinolytic enzyme,LUFE)在大鼠动脉血栓形成过程中对血管内膜损伤的保护作用。方法:将50 只SD大鼠随机分为假手术组、模型组、阿司匹林组和LUFE低、高剂量组。建立血栓模型前,阿司匹林组和LUFE低、高剂量组分别以10 mg/kg mb阿司匹林和100、400 mg/kg mb LUFE灌胃7 d。次日,除假手术组以外其余4 组以三氯化铁诱导建立大鼠颈动脉血栓模型。苏木精-伊红(hematoxylin-eosin,HE)染色观察颈动脉血管形态学变化;酶联免疫吸附法测定血清C反应蛋白(C-reactive protein,CRP)、白细胞介素(interleukin,IL)-6、IL-8、单核细胞趋化蛋白-1(monocyte chemoattractant protein-1,MCP-1)、细胞间黏附分子-1(intercellular cell adhesion molecule-1,ICAM-1)和血小板-内皮细胞黏附分子-1(platelet endothelial cell adhesion moleculer-1,PECAM-1)水平;采用流式细胞仪测定血小板-白细胞聚集体(platelet-leukocyte aggregates,PLA)水平。在体外,经LUFE干预后观察血小板在固化胶原上的黏附作用;采用流式细胞仪测定胶原刺激后血小板Ca2+水平;Western blot方法观察血小板T细胞跨膜连接蛋白(linker for activation of T cells,LAT)和磷脂酶Cγ(phospholipase Cγ,PLCγ)磷酸化水平。结果:LUFE干预后能明显减轻血管内膜的损伤,不同程度地降低血栓模型大鼠血清CPR、IL-6、IL-8、MCP-1、ICAM-1和PECAM-1水平,降低PLA水平。在体外,LUFE能降低血小板与胶原的黏附,并降低血小板Ca2+水平,下调血小板LAT和PLCγ蛋白的磷酸化水平。结论:LUFE抑制血栓形成过程中的血管炎症反应,这可能与其血管内皮细胞保护作用和抗血小板作用有关。

关键词: 榆干离褶伞;溶栓酶;血栓;炎症;血小板

Abstract: Objective: To study the protective effect of Lyophyllum ulmarium fibrinolytic enzyme (LUFE) on intimal injury during arterial thrombosis in rats. Methods: Fifty SD rats were randomly divided into five groups: sham operation, model, aspirin, and low- and high-dose LUFE. Before the model establishment, the rats in the aspirin and LUFE groups were administrated with 10 mg/kg mb of aspirin, and 100 and 400 mg/kg mb of LUFE, respectively, by gavage for seven consecutive days. On the following day, the rats in all groups except the sham operation group were administered with ferric chloride to induce carotid artery thrombosis. The morphological changes of carotid artery were observed by HE staining. The serum levels of C-reactive protein (CRP), interleukin (IL)-6, IL-8, monocyte chemoattractant protein-1 (MCP-1), intercellular cell adhesion molecule-1 (ICAM-1), and platelet endothelial cell adhesion molecule-1 (PECAM-1) were detected by an enzyme-linked immunosorbent assay. The level of platelet-leukocyte aggregates (PLA) was detected by flow cytometry. The adhesion of platelets to solidified collagen was observed in vitro after LUFE intervention. The level of Ca2+ in platelets stimulated by collagen was detected by flow cytometry. The phosphorylation levels of platelet linker for activation of T cells (LAT) and phospholipase Cγ (PLCγ) were detected by Western blot. Results: LUFE intervention significantly reduced vascular intimal injury, lowered the levels of serum CPR, IL-6, IL-8, MCP-1, ICAM-1 and PECAM-1 in different degrees, and decreased the level of PLA. LUFE reduced in vitro platelet adhesion to collagen, decreased platelet Ca2+ level, and down-regulated the phosphorylation of platelet LAT and PLCγ proteins. Conclusion: LUFE inhibits vascular inflammation during thrombosis, which may be related to the protective effect on vascular endothelial cells and the antiplatelet effect of LUFE.

Key words: Lyophyllum ulmarium; fibrinolytic enzyme; thrombus; inflammation; platelets

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