TLR4mAb抑制mmLDL损伤小鼠肠系膜动脉内皮依赖性舒张功能

陈根, 林杰, 秦旭平, 周楠, 李洁

中国药学杂志 ›› 2019, Vol. 54 ›› Issue (4) : 274-283.

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中国药学杂志
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中国药学杂志 ›› 2019, Vol. 54 ›› Issue (4) : 274-283. DOI: 10.11669/cpj.2019.04.005
论著

TLR4mAb抑制mmLDL损伤小鼠肠系膜动脉内皮依赖性舒张功能

  • 陈根1,2, 林杰1, 秦旭平2, 周楠3, 李洁1,2*
作者信息 +

Inhibition Effect of TLR4mAb on mmLDL Impaired Endothelium-dependent Vasodilatation in Mouse Mesenteric Artery

  • CHEN Gen1,2, LIN Jie1, QIN Xu-ping2, ZHOU Nan3, LI Jie1,2*
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文章历史 +

摘要

目的 探讨预先使用TLR4单克隆抗体(TLR4mAb)对轻度氧化修饰低密度脂蛋白(mmLDL)诱发小鼠肠系膜动脉内皮依赖性舒张功能损伤的影响及作用机制。方法 实验分为:空白对照组、mmLDL处理组、TLR4mAb干预组。采用ELISA方法测定血浆中白细胞介素(IL)-1β和肿瘤坏死因子(TNF)-α的浓度水平,微血管张力描记技术测定血管内皮依赖性舒张功能,Western blot技术和RT-PCR技术分别考察血管组织蛋白表达量和mRNA表达水平,透射电镜观察肠系膜动脉内皮细胞超显微结构。结果 TLR4mAb剂量依赖性改善mmLDL损伤血管内皮依赖性舒张功能的损伤作用,显著上调KCa3.1-通道、KCa2.3-通道蛋白表达和下调炎症因子TNF-α和IL-1β表达。TLR4mAb改善mmLDL损伤血管内皮细胞及内皮依赖性舒张功能,可能与其竞争性拮抗mmLDL激活TLR4信号转导通路及其下游的NF-κBp65和p-38MAPK通路有关。结论 预先使用TLR4mAb可以减轻mmLDL所诱发的内皮细胞损伤和内皮依赖性舒张功能降低,以及抑制炎症因子的过度表达,调控TLR4通路及其下游的NF-κBp65和p-38MAPK通路可能是预防治疗心血管疾病的有效靶点。

Abstract

OBJECTIVE To investigate the effect and mechanism of TLR4 monoclonal antibody (TLR4mAb) on mmLDL induced impairment of endothelium-dependent vasodilatation in mouse mesenteric artery. METHODS The experiment established three groups of normal saline group, mmLDL treatment group and TLR4mAb intervention group. The concentration of IL-1β and TNF-α in plasma was determined by enzyme-linked immunosorbent assay (ELISA). Measurement of endothelium-dependent vasodilatation was achieved by microvascular tension mapping. Western blot and RT-PCR were used to investigate the expression level of protein and mRNA expressions in vascular tissues. In addition, ultra-structure of mesenteric artery endothelial cells was observed by transmission electron microscope. RESULTS TLR4mAb could improve the damage of mmLDL induced impairment of endothelium-dependent vasodilatation in a dose-dependent manner. Besides, TLR4mAb obviously up-regulated protein expressions in KCa3.1-channel and KCa2.3-channel, and down-regulated the expression of inflammatory factors TNF-α and IL-1β. Furthermore, the improvement of mmLDL impaired vascular endothelial cells and endothelium-dependent vasodilatation might be correlated with its competitive antagonism of mmLDL-activated TLR4 signal transduction pathway and its downstream NF-κBp65 and p-38 MAPK pathway. CONCLUSION Administration of TLR4mAb in advance can alleviate the impairment of endothelial cells and the decrease of endothelium-dependent vasodilatation induced by mmLDL, and inhibit the overexpression of inflammatory factors. Regulation of TLR4 pathway as well as its downstream NF-κBp65 and P-38 MAPK pathways may be effective targets for the prevention and treatment of cardiovascular diseases.

关键词

TLR4单克隆抗体 / 轻度氧化修饰低密度脂蛋白 / 炎症 / 肠系膜动脉 / 内皮依赖性舒张

Key words

toll like receptor4 monoclonal antibodies / minimally modified low density lipoprotein / inflammation / mesenteric artery / endothelium dependent relaxation

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陈根, 林杰, 秦旭平, 周楠, 李洁. TLR4mAb抑制mmLDL损伤小鼠肠系膜动脉内皮依赖性舒张功能[J]. 中国药学杂志, 2019, 54(4): 274-283 https://doi.org/10.11669/cpj.2019.04.005
CHEN Gen, LIN Jie, QIN Xu-ping, ZHOU Nan, LI Jie. Inhibition Effect of TLR4mAb on mmLDL Impaired Endothelium-dependent Vasodilatation in Mouse Mesenteric Artery[J]. Chinese Pharmaceutical Journal, 2019, 54(4): 274-283 https://doi.org/10.11669/cpj.2019.04.005
中图分类号： R965   

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基金

国家自然科学基金项目资助(81202535);湖南省科技厅自然科学基金项目资助(2018JJ6097);湖南省卫生计生委课题资助(B2017183);郴州市科技局项目资助(jsyf2017024,jsyf2017025)
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