Effect of Essential Oil of Fructus  in Inhibiting Inflammatory Injury of Human Aortic Endothelial Cells Induced by Lipopolysaccharide by Regulating NF-B Signaling

LIN Dan; SHI Ting-yu; YANG Hong; WU Guo-ping; FU Ling-yun; TAO Ling; SHEN Xiang-chun

doi:10.13422/j.cnki.syfjx.20181929

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Effect of Essential Oil of Fructus in Inhibiting Inflammatory Injury of Human Aortic Endothelial Cells Induced by Lipopolysaccharide by Regulating NF-B Signaling

更新时间：2024-01-04

- Effect of Essential Oil of Fructus in Inhibiting Inflammatory Injury of Human Aortic Endothelial Cells Induced by Lipopolysaccharide by Regulating NF-B Signaling
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 19, Pages: 139-144(2018)
- 作者机构：
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- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20181929
  CLC： R285.5
- Published：2018
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LIN Dan, SHI Ting-yu, YANG Hong, et al. Effect of Essential Oil of Fructus in Inhibiting Inflammatory Injury of Human Aortic Endothelial Cells Induced by Lipopolysaccharide by Regulating NF-B Signaling[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(19): 139-144.
DOI：

LIN Dan, SHI Ting-yu, YANG Hong, et al. Effect of Essential Oil of Fructus in Inhibiting Inflammatory Injury of Human Aortic Endothelial Cells Induced by Lipopolysaccharide by Regulating NF-B Signaling[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(19): 139-144. DOI： 10.13422/j.cnki.syfjx.20181929.

摘要

目的：研究艳山姜挥发油（essential oil of Alpinia zerumber fructus，EOAZF）对脂多糖（lipopolysaccharide，LPS）诱导人主动脉内皮细胞（human aortic endothelial cells，HAECs）炎性损伤的保护作用。方法：体外传代培养HAECs，先用不同剂量EOAZF（0.05，0.1 mg·L-1）和阿司匹林（0.5 mmol·L-1）预处理1 h，另设空白组，再与LPS（5 mg·L-1）共同作用12 h。采用噻唑蓝（MTT）比色法检测细胞活性；苏木素-伊红染色进行细胞形态学观察；细胞划痕修复实验分析HAECs划痕修复能力；蛋白免疫印迹法（Western blot）检测血管细胞黏附分子-1（vascular cell adhesion molecule-1，VCAM-1），半胱氨酸天冬氨酸蛋白酶-3（Caspase-3），Toll样受体4（Toll-like receptor 4，TLR4），核转录因子-κB抑制蛋白激酶（inhibitor of nuclear factor kappa-B kinase，IKK），磷酸化NF-κB抑制蛋白激酶（phospho-inhibitor of nuclear factor kappa-B kinase，p-IKK）蛋白的表达；实时荧光定量聚合酶链式反应（Real-time PCR）检测细胞中TLR4和VCAM-1 mRNA的表达；免疫荧光法观察NF-κB p65的核转位情况。结果：与空白组比较，LPS能诱导HAECs形态学损伤，降低细胞存活率及划痕修复能力，同时上调VCAM-1，Caspase-3，TLR4，IKK及P-IKK的表达。免疫荧光实验结果分析表明LPS能够诱导NF-κB p65的核转位。与模型组比较，EOAZF预处理后，能够改善LPS诱导的HAECs形态学损伤，提高细胞存活率及划痕修复能力，抑制VCAM-1，Caspase-3，TLR4，IKK及P-IKK的表达及NF-κB p65的核转位。结论：EOAZF对LPS诱导的HAECs炎性损伤具有保护作用，其机制与调控TLR4/IKK/NF-κB-p65信号有关。

Abstract

Objective: To study the protective effect of essential oil of Alpinia zerumber fructus(EOAZF) on the inflammatory injury of human aortic endothelial cells (HAECs) induced by lipopolysaccharide (LPS). Method: HAECs were cultured in vitro. HAECs were pre-treated with different doses of EOAZF (0.05

0.1 mg·L-1) and aspirin (0.5 mmol·L-1) for 1 h

then incubated with lipopolysaccharide(LPS) (5 mg·L-1) for 12 h. The effect of EOAZF on the cell viability of HAECs induced by LPS was detected by methyl thiazolyl tetrazolium (MTT) assay

hematoxylin eosin(HE) staining was used to observe the cell morphology

and wound healing assay was used to analyze the ability of HAECs wound healing ability. Western blot was used to detect the protein expressions of vascular cell adhesion molecule-1 (VCAM-1)

cysteine aspartase-3(Caspase-3)

Toll like receptor 4 (TLR4)

nuclear factor kappa B inhibitory protein kinase (IKK) and phosphorylated nuclear factor kappa inhibitory protein kinase (p-IKK)

and real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expressions of TLR4 and VCAM-1.The nuclear translocation of nuclear factor kappa-B p65(NF-κB p65) was observed by immunofluorescence. Result: Compared with control group

LPS induced HAECs morphological damage

decreased cell viability and wound healing ability

and increased the expressions of VCAM-1

Caspase-3

TLR4

IKK and p-IKK. Immunofluorescence results showed that LPS induced nuclear translocation of NF-κB p65. Compared with model group

the pre-treatment of EOAZF improved the morphological damage of HAECs induced by LPS

increased cell viability and wound healing ability

inhibited the expressions of VCAM-1

Caspase-3

TLR4

IKK and p-IKK

as well as significantly inhibited the nuclear transposition of NF-κB p65. Conclusion: EOAZF has the protective effect on the inflammatory injury of HAECs induced by LPS

and its mechanism may be related to the regulation of TLR4/IKK/NF-κB p65 signaling.

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