消溶稳斑方对ox-LDL诱导的人脐静脉内皮细胞炎症损伤的作用研究-《西部中医药》

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Title:: Study on the Effects of Plaque Dissolving and Stablilizing Prescription on Inflammatory Injury of Human Umbilical Vein Endothelial Cells Induced by ox-LDL

作者:: 蒋静¹, 邵静²; 1.河南中医药大学，河南郑州 450000
2.河南中医药大学第一临床医学院

Author(s):: JIANG Jing¹, SHAO Jing²; 1.Henan University of TCM, Zhengzhou 450000, China
2.The First Clinical Medical College, Henan University of TCM

关键词:: 消溶稳斑方; 人脐静脉内皮细胞; 动脉粥样硬化; 炎症反应; AMPK/SIRT1通路

Keywords:: plaque dissolving and stabilizing prescription; human umbilical vein endothelial cells; atherosclerosis; inflammatory reaction; AMPK/SIRT1 pathway

分类号:: R543.1+2

DOI:: 10.12174/j.issn.2096-9600.2021.10.05

摘要:: 目的通过探讨消溶稳斑方对氧化低密度脂蛋白（oxidation low lipoprotein，ox-LDL）诱导的人脐静脉内皮细胞炎症反应的作用，进一步阐述消溶稳斑方对炎症反应诱发的动脉粥样硬化的治疗作用与机制。方法选择培养的第三代人脐静脉内皮细胞，将其分为空白组，模型组，他汀组，中药低、中、高浓度组，中药中浓度+抵抗素组，给予对应药物干预。采用蛋白质印迹法（Western blot）测定各组单磷酸腺苷酶活化蛋白激酶（amp-activated protein kinas，p-AMPK）、沉默信息调节因子1（silent information regulator 1，SIRT1）水平，采用逆转录-聚合酶链反应（reverse transcriptase-polymerase chain reaction，RT-PCR）技术检测抑制蛋白激酶β（（inhibitor kappa B kinaseβ，IKKβ）mRNA、基质金属蛋白酶9（matrix metalloproteinase-9，MMP-9）mRNA表达，比较各组之间的差异。结果与空白组比较，模型组，他汀组，中药低、中浓度组，中药中浓度+抵抗素组p-AMPK、SIRT1水平均不同程度降低（P＜0.05），IKKβ mRNA、MMP-9 mRNA表达呈上升趋势（P＜0.05）。与模型组比较，他汀组，中药低、中、高浓度组p-AMPK、SIRT1水平均升高（P＜0.05），IKKβ mRNA、MMP-9 mRNA表达呈下降趋势（P＜0.05）。与他汀组比较，中药低浓度组p-AMPK、SIRT1水平降低（P＜0.05），中药中、高浓度组p-AMPK、SIRT1水平升高，但差异无统计学意义（P＞0.05），IKKβ mRNA、MMP-9 mRNA表达呈下降趋势（P＜0.05）。结论消溶稳斑方可通过作用于AMPK/SIRT1信号通路，上调p-AMPK、SIRT1水平，抑制IKKβ mRNA、MMP-9 mRNA表达，阻断炎症因子释放，抑制炎症反应造成的血管内皮细胞损伤，发挥保护血管内皮细胞、防治动脉粥样硬化的作用。

Abstract:: ObjectiveTo further elucidate therapeutic effects and the mechanism of plaque dissolving and stabilizing prescription for inflammation-induced atherosclerosis through exploring the effects of the prescription on inflammatory reaction of human umbilical vein endothelial cells induced by ox-LDL. MethodsThe third generation of human umbilical vein endothelial cells were chosen, cultured and divided into the blank group, the model group, statin group, low, moderate and high concentrations groups of herbs, moderate concentrations of herbs+resistin group, which were intervened with the corresponding medicine. Western blot method was used to detect the levels of p-AMPK and SIRT1 in different groups, RT-PCR was used to measure the expressions of IKKβ mRNA and MMP-9 mRNA, and compare the difference between different groups. ResultsCompared with the blank group, the levels of p-AMPK and SIRT1 lowered in the model group, statin group, low and moderate concentrations groups of herbs, moderate concentrations of herbs+resistin group (P<0.05), the expressions of IKKβ mRNA and MMP-9 mRNA were on the rise (P<0.05). Compared with the model group, the levels of p-AMPK and SIRT1 increased in statin group, low, moderate and high concentrations groups of herbs (P<0.05), the expressions of IKKβ mRNA and MMP-9 mRNA presented a downward trend (P<0.05). Compared with statin group, the levels of p-AMPK and SIRT1 reduced in low concentration group of herbs (P<0.05), and rose in moderate and high concentrations groups of herbs, while the difference had no statistical meaning (P>0.05), the expressions of IKKβ mRNA and MMP-9 mRNA showed a downward trend (P<0.05). ConclusionPlaque dissolving and stabilizing prescription could protect vascular endothelial cell, prevent and treat atherosclerosis through acting on AMPK/SIRT1 signaling pathway, up regulating the levels of p-AMPK and SIRT1, inhibiting the expressions of IKKβ mRNA and MMP-9 mRNA, blocking the release of inflammatory factors and restraining vascular endothelial cell injury caused by inflammatory reaction.

《西部中医药》[ISSN:2096-9600/CN:62-1204/R]

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