灯盏花素对急性肺栓塞的血管内皮保护作用及抗炎机制研究OA
Study on the Protective Effect of Breviscapine on Vascular Endothelium and Its Anti-inflammatory Mechanism in Acute Pulmonary Embolism
目的:探究灯盏花素(breviscapine,Bre)对急性肺栓塞(acute pulmonary embolism,APE)血管内皮保护作用及抗炎机制.方法:将SD大鼠随机分为假手术(Sham)组、APE组、APE+低分子肝素钙注射液(APE+LMWH)组、APE+灯盏花素低剂量(APE+Bre-L)组和APE+灯盏花素高剂量(APE+Bre-H)组(n=6).采用自体血栓构建APE大鼠模型.评估平均肺动脉压(mPAP)、右心室压力(RVP)、动脉血氧分压(PaO2)与动脉血二氧化碳分压(PaCO2),HE染色观察肺组织病理损伤.此外,采用凝血酶刺激人肺动脉内皮细胞(human pul-monary artery endothelial cells,HPAECs)构建体外细胞模型,并给予Bre和TLR-4激活剂LPS干预.CCK8法测定细胞活力.ELISA法测定大鼠血清D-二聚体(D-D)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、单核细胞趋化蛋白-1(MCP-1)、内皮素-1(ET-1)、血栓素B2(TXB2)水平,以及细胞血管性血友病因子(vWF)、血管内皮细胞黏附分子-1(VCAM-1)和P选择素含量.免疫荧光法测定HPAECs中NF-κB p65的核易位情况.Western blot法检测TLR-4/MyD88/NF-κB通路蛋白表达.结果:相较于Sham组,APE组大鼠肺组织出现严重病理损伤,mPAP、RVP、D-D、TNF-α、IL-1β、MCP-1、ET-1、TXB2、TLR-4、MyD88与p-NF-κB/NF-κB的表达均升高,而PaO2水平降低.相较于APE组大鼠,APE+LMWH组和APE+Bre-H组的上述指标得到改善.在凝血酶刺激的HPAECs中,Bre提高细胞活力,降低vWF、VCAM-1和P选择素含量,抑制NF-κB p65的核易位,并下调TLR-4、MyD88与p-NF-κB/NF-κB的表达.此外,TLR-4激活剂LPS削弱了Bre的以上效应.结论:Bre可通过改善APE的炎症反应和血管内皮功能障碍,改善肺血气功能和肺血流动力学,缓解肺动脉高压和低氧血症以减缓APE的发生进展,这一过程可能与TLR-4/MyD88/NF-κB信号通路的抑制密切相关.本研究为Bre干预在APE治疗中的临床应用提供了坚实的理论基础.
Objective:To investigate the protective effect and anti-inflammatory mechanism of breviscapine(Bre)on vascular endothelium in acute pulmonary embolism(APE).Methods:SD rats were randomly divided into Sham group,APE group,APE+low molecular weight heparin calcium injection(APE+LMWH)group,APE+breviscapine low-dose(APE+Bre-L)group,and APE+breviscapine high-dose(APE+Bre-H)group.APE rat models were constructed using autologous thrombus.Mean pulmonary artery pressure(mPAP)and right ventricular pressure(RVP)were measured using an electrocardiogram monitor,and arterial oxygen partial pressure(PaO2)and arterial carbon dioxide partial pressure(PaCO2)were measured.Lung tissue pathologi-cal damage was examined by HE staining.Human pulmonary artery endothelial cells(HPAECs)were stimu-lated with thrombin to construct an in vitro cell model,followed by intervention with Bre and TLR4 agonist LPS.Cell viability,serum dimer(DD),interleukin-1 β(IL-1 β),tumor necrosis factor-α(TNF-α),monocyte chemoattractant protein-1(MCP-1),endothelin-1(ET-1),thromboxane B2(TXB2)levels,and von Willebrand factor(vWF),vascular endothelial cell adhesion molecule-1(VCAM-1),and P-selectin levels in HPAECs were assessed using CCK8 and ELISA.Nuclear translocation of NF-κ B p65 in HPAECs was determined by immunofluorescence assay.The expression of TLR-4/MyD88/NF-κB pathway proteins was de-termined by Western blot.Results:Compared with the Sham group,APE rats showed severe pathological damage,with increased mPAP,RVP,DD,TNF-α,IL-1 β,MCP-1,ET-1,TXB2,TLR-4,MyD88,and p-NF-κ B/NF-κ B expression,while decreased PaO2 levels.LMWH and Bre-H treatments in APE rats improved the above-mentioned indicators.In HPAECs stimulated by thrombin,Bre increased cell viability,reduced vWF,VCAM-1,and P-selectin levels,inhibited nuclear translocation of NF-κ B p65,and down-regulated TLR-4,MyD88,and p-NF-κ B/NF-κ B expression.In addition,LPS weakened the above ef-fects.Conclusion:Bre may alleviate the inflammatory response and endothelial dysfunction to improve APE,which may be closely related to the inhibition of TLR-4/MyD88/NF-κ B pathway.
张言涛;吴丽丽;周黎明
杭州市中医院 浙江 杭州 310007杭州市中医院 浙江 杭州 310007杭州市中医院 浙江 杭州 310007
灯盏花素急性肺栓塞炎症反应内皮功能障碍TLR-4/MyD88/NF-κB通路
breviscapineacute pulmonary embolisminflammatory responseendothelial dysfunctionTLR-4/MyD88/NF-κ B pathway
《浙江中医杂志》 2026 (3)
189-194,6
浙江省中医药科技计划项目(2024ZL661)
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