藁本内酯上调miR⁃181b 减轻氧化低密度脂蛋白诱导人脐静脉内皮细胞炎症损伤
作者:
作者单位:

(1. 南方医科大学第三附属医院耳鼻咽喉头颈外科,广州 510630; 2. 南方医科大学中医药学院,广州 510515; 3. 广东省人民医院中医科,广州 510080)

中图分类号:

R-33

基金项目:

国家自然科学基金项目(编号:81774213,81403339);广东省中管局课题(编号:81373810)?


Ligustilide alleviates inflammatory reactions induced by ox⁃LDL in HUVECs through upregulation of miR⁃181b
Author:
Affiliation:

(1. Department of E. N. T. , the Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, China. 2. College of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515 3. Department of Traditional Chinese Medicine, Guangdong General Hospital, Guangzhou 510080)

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    摘要:

    目的 研究藁本内酯(ligustilide,LIG)减轻氧化低密度脂蛋白(oxidative low density lipoprotein,ox?LDL)诱导人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)损伤作用机制?方法 将细胞随机分为5 组,对照组(正常培养细胞不加干预)?模型组(100 μmol/ L ox?LDL 干预24 h)?LIG 低浓度组?LIG 中浓度组?LIG 高浓度组?其中,LIG 低?中?高浓度组细胞分别以10?20?40 μmol/ L 预先孵育2 h,再用100 μmol/ L ox?LDL 干预24 h?CCK8 法检测各组细胞活力,ELISA 检测细胞培养液中肿瘤坏死因子(TNF?α)?白细胞介素6(IL?6)?细胞间黏附分子1(ICAM?1)和血管细胞粘附分子?1(VCAM?1)的含量?qRT?PCR 检测细胞内TNF?α?IL?6?ICAM?1?VCAM?1?NF?κB 及miR?181b 的mRNA 水平;Western blotting 测定细胞NF?κB 的表达量?结果 与对照组相比,模型组细胞存活率明显下降,凋亡率增高,细胞上清中TNF?α?IL?6?ICAM?1 及VCAM?1 的浓度升高?LIG 能显著提高细胞存活率,抑制细胞凋亡,并且能降低细胞内炎性因子水平,尤其是中?高浓度组,与模型组相比差异有显著性( P < 0.01)?此外,LIG 能明显降低细胞内NF?κB 转录及翻译水平,上调miR?181b 的表达,与模型组相比差异有显著性( P < 0.01)?激活或抑制细胞内miR?181b 表达,能有效调节细胞内炎性因子水平?结论 藁本内酯明显减轻ox?LDL 诱导HUVEC 损伤,其机制可能是通过上调miR?181b 抑制NF?κB 表达,从而减轻ox?LDL 诱导的细胞炎症损伤?

    Abstract:

    Objective To study the molecular mechanism of ligustilide (LIG) in HUVEC injury induced by ox?LDL. Methods HUVECs were randomly divided into five groups: control group (untreated cells), model group (cells treated with 100 μmol/ L ox?LDL for 24 h), and LIG groups (cells pre?treated with 10, 20, or 40 μmol/ L LIG for 2 h and then exposed to 100 μmol/ L ox?LDL for 24 h). Cell viability was measured by CCK8 assays. Levels of inflammatory cytokines TNF?α and IL?6, and ICAM?1 and VCAM?1 were assayed by ELISAs. mRNA levels of TNF?α, IL?6, ICAM?1, VCAM?1, and miR?181b were determined by qRT?PCR and the protein level of NF?κB by Western blotting. Results Compared with the control group, ox?LDL induced apoptosis and inflammatory cytokine secretion in HUVECs ( P <0.01). LIG protected cells and reduced apoptosis. Moreover, LIG reduced the secretion of inflammatory cytokines compared with the model group ( P < 0.01). LIG also upregulated miR?181b expression and downregulated its target gene, NF?κB. Inhibition of miR?181b mostly reduced the effect of LIG on inflammation in HUVECs. Conclusions LIG reduces damage by ox?LDL in HUVECs, partly through inhibiting expression of NF?κB via miR?181b to reduce the inflammatory reaction induced by ox?LDL.

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黄志勇,田广永,曹玲,詹坚宏,赵丹丹,于云红,周凤华.藁本内酯上调miR⁃181b 减轻氧化低密度脂蛋白诱导人脐静脉内皮细胞炎症损伤[J].中国比较医学杂志,2018,28(7):18~23.

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  • 收稿日期:2018-04-12
  • 在线发布日期: 2018-08-08
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