榆干离褶伞溶栓酶对大鼠动脉血栓的抑制作用

doi:10.7506/spkx1002-6630-20200118-226

摘要/Abstract

摘要： 目的：研究榆干离褶伞溶栓酶（Lyophyllum ulmarium fibrinolytic enzyme，LUFE）对大鼠动脉血栓形成的抑制作用及其机制。方法：将SD大鼠随机分为假手术组、模型组、阳性对照组和LUFE低、高剂量组，除假手术组以外，其余4 组均以三氯化铁建立颈动脉血栓模型。造模前阳性对照组和LUFE低、高剂量组分别以10 mg/kg mb阿司匹林以及100、400 mg/kg mb LUFE灌胃7 d。观察血栓质量和颈动脉血管组织病理变化；酶联免疫吸附测定法检测肿瘤坏死因子α（tumor necrosis factor alpha，TNF-α）、白细胞介素-6（interleukin-6，IL-6）、血小板活化因子（platelet-activating factor，PAF）、可溶性血栓调节蛋白（soluble thrombomodulin，sTM）、E-选择素和组织因子（tissue factor，TF）水平；流式细胞仪检测血小板CD62P表达水平。为观察LUFE对血小板功能的影响，在体外以胶原诱导血小板活化，观察LUFE对大鼠富血小板血浆中P-选择素、β-血小板球蛋白（β-thromboglobulin，β-TG）和血小板膜糖蛋白IIbIIIa（glycoprotein IIbIIIa，GPIIbIIIa）水平的影响；Western blot方法观察LUFE对大鼠血小板磷脂酰肌醇3激酶（phosphatidylinositol 3 kinase，PI3K）、蛋白激酶B（protein kinase B，Akt）和p38丝裂原活化蛋白激酶（p38 mitogen-activated protein kinase，p38 MAPK）表达水平的影响。结果：LUFE能明显减轻血栓质量和血管受损程度，抑制血栓模型大鼠血清sTM、E-选择素、TF、TNF-α、IL-6和PAF水平的上调，下调血小板CD62P的表达。同时，在体外LUFE能够下调胶原诱导的富血小板血浆中P-选择素、β-TG和GPIIbIIIa水平，下调血小板PI3K、Akt和p38 MAPK蛋白的磷酸化水平。结论：LUFE具有抗血栓作用，其机制可能与LUFE对血管内皮细胞的保护作用以及对血小板活化的抑制作用有关。

关键词: 榆干离褶伞；溶栓酶；抗血栓作用；血管内皮细胞；血小板

Abstract: Objective: To explore the inhibitory effect of the fibrinolytic enzyme from Lyophyllum ulmarium (LUFE) on rat thrombosis. Methods: SD rats were randomly divided into sham operation, model, positive control and low- and high-dose LUFE groups. All groups except the sham operation group were used to establish a ferric chloride (FeCl3)-induced carotid arterial thrombosis model. Before modeling, the rats in the positive control group and the low- and high-dose LUFE groups were administered with 10 mg/kg mb of aspirin and 100 and 400 mg/kg mb of LUFE, respectively, by gavage for 7 days. The changes in thrombus mass and carotid vascular histopathology were observed. The levels of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), platelet-activating factor (PAF), soluble thrombomodulin (sTM), E-selectin and tissue factor (TF) were detected by enzyme-linked immunosorbent assay. The expression level of CD62P in platelets was detected by flow cytometry. In order to observe the effect of LUFE on platelet function, collagen-induced platelet activation was performed in vitro to observe the effect of LUFE on P-selectin, β-thromboglobulin (β-TG) and platelet membrane glycoprotein IIbIIIa (GPIIbIIIa) levels in platelet-rich plasma. Western blot was used to evaluate the effect of LUFE on the expression levels of phosphatidylinositol 3 kinase (PI3K), protein kinase B (Akt) and p38 mitogen-activated protein kinase (p38 MAPK) in platelets. Results: LUFE significantly reduced thrombus mass and vascular damage, inhibited the increase in serum sTM, E-selectin, TF, TNF-α, IL-6 and PAF levels in the thrombus model rats, and decreased the expression of platelet CD62P. LUFE could in vitro reduce the levels of P-selectin, β-TG and GPIIbIIIa in collagen induced platelet-rich plasma, and down-regulated the phosphorylation levels of platelet PI3K, Akt and p38 MAPK protein. Conclusion: LUFE has an anti-thrombotic effect, and its mechanism may be related to the protection of vascular endothelial cells and the inhibition of platelet activation.

Key words: Lyophyllum ulmarium; thrombolytic enzyme; antithrombotic effect; vascular endothelial cells; platelets

中图分类号:

R285.5

李芳芳，卫莹，刘雪，沈明花. 榆干离褶伞溶栓酶对大鼠动脉血栓的抑制作用[J]. 食品科学, 2021, 42(1): 180-186.

LI Fangfang, WEI Ying, LIU Xue, SHEN Minghua. Antithrombotic Effect of Fibrinolytic Enzyme from Lyophyllum ulmarium on Arterial Thrombosis in Rats[J]. FOOD SCIENCE, 2021, 42(1): 180-186.

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榆干离褶伞溶栓酶对大鼠动脉血栓的抑制作用

Antithrombotic Effect of Fibrinolytic Enzyme from Lyophyllum ulmarium on Arterial Thrombosis in Rats

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