雷公藤甲素对血管内皮细胞炎症反应的影响及机制研究  

The mechanisms for the effect of triptolide on inflammatory response in endothelial cells

作　　者：张诗雨 张静[1] 高攀 刘素晓 崔琳[2] 闫凤娜 高原[2] 谢世阳[2] 朱明军[2] 王幼平[2] ZHANG Shi-yu;ZHANG Jing;GAO Pan;LIU Su-xiao;CUI Lin;YAN Feng-na;GAO Yuan;XIE Shi-yang;ZHU Ming-jun;WANG You-ping(Henan University of Chinese Medicine,Zhengzhou,Henan,450000,China;First Affiliated Hospital,Henan University of Chinese Medicine,Zhengzhou,Henan,450000,China)

机构地区：[1]河南中医药大学,河南郑州450000 [2]河南中医药大学第一附属医院,河南郑州450000

出　　处：《时珍国医国药》2021年第7期1537-1541,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81673734,82074194)。

摘　　要：目的通过人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVECs)培养模型,探明雷公藤甲素对血管内皮细胞在内毒素(lipopolysaccharide, LPS)刺激下产生炎性介质的影响,以及在该病理条件下对血管内皮细胞核转录因子nuclear factor-κB(NF-κB)的影响,从而阐明雷公藤甲素抑制炎症反应的分子生物学机制。方法通过HUVECs培养模型,利用ELISA、细胞酶联反应法、免疫荧光标记的单核细胞粘附实验及核转录因子NF-κB活性测定等方法,在LPS刺激条件下,观察雷公藤甲素对血管内皮细胞释放炎性介质、单核细胞粘附于血管内皮细胞及核转录因子NF-κB激活程度的影响。结果与正常对照组相比,LPS明显促进血管内皮细胞产生释放细胞因子TNF-α[(170.1±17.3) pg·mL^(-1)]和IL-6 [(58.1±6.7) pg·mL^(-1)]、趋化因子MCP-1 [(2152.8±211.5) pg·mL^(-1)]及粘附因子ICAM-1 (4.00±0.51)和VCAM-1 (3.56±0.33)的表达(P<0.05),同时LPS显著促进单核细胞THP-1粘附于血管内皮细胞(409.3±42.8)(P<0.05),以及血管内皮细胞IκBα的磷酸化(3.39±0.27)和核转录因子NF-κB的激活(6.96±0.94)(P<0.05),而雷公藤甲素(25 nmol·L^(-1),50 nmol·L^(-1),100 nmol·L^(-1))明显地抑制LPS所诱发的上述实验结果 (P<0.05),并呈剂量依赖性。结论雷公藤甲素通过抑制NF-κB信号通路减轻和缓解血管内皮细胞炎症反应,从而实现其抑制炎症反应的作用。Objective This study was designed to determine the effects and mechanisms of triptolide on lipopolysaccharide(LPS)-induced release of inflammatory mediators and activation of NF-κB signaling in human umbilical vein endothelial cells(HUVECs). Methods In cultured HUVECs, we observed the effects of triptolide on LPS-mediated release of inflammatory mediators, monocyte adhesion, and activated NF-κB signaling by the use of ELISA, cell-based ELISA, fluorescently labeled monocyte adhesion, and NF-κB activity assay. Results In HUVECs, treatment with LPS increased the release of cytokine TNF-α [(170.1 ± 17.3) pg·mL^(-1)], IL-6 [(58.1 ± 6.7) pg·mL^(-1)], chemokine MCP-1 [(2152.8 ± 211.5) pg·mL^(-1)], and the expression of adhesion molecule ICAM-1(4.00 ± 0.51) and VCAM-1(3.56 ± 0.33)(P<0.05). These were accompanied by increased THP-1 adhesion(409.3 ± 42.8)(P<0.05), and the phosphorylation of IκBα(3.39 ± 0.27) and activation of NF-κB(6.96 ± 0.94)(P<0.05). In addition, we found that these effects were attenuated by triptolide(25 nmol·L^(-1), 50 nmol·L^(-1), 100 nmol·L^(-1)) in a dose-dependent manner(P<0.05). Conclusion Our data indicate that triptolide attenuates the LPS-induced inflammatory responses of endothelial cells via suppression of NF-κB signaling pathway, leading to its anti-inflammation.

关 键 词：雷公藤甲素 内毒素 血管内皮细胞 炎症 核转录因子NF-ΚB 

分 类 号：R285.5[医药卫生—中药学]