Full Text:   <3202>

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CLC number: O629.9

On-line Access: 2016-06-03

Received: 2015-09-04

Revision Accepted: 2015-12-18

Crosschecked: 2016-05-12

Cited: 4

Clicked: 5976

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Jiu-liang Zhang

http://orcid.org/0000-0002-1745-846X

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Journal of Zhejiang University SCIENCE B 2016 Vol.17 No.6 P.425-436

http://doi.org/10.1631/jzus.B1500213


Inhibitory effect of Gardenblue blueberry (Vaccinium ashei Reade) anthocyanin extracts on lipopolysaccharide-stimulated inflammatory response in RAW 264.7 cells


Author(s):  Wei Xu, Qing Zhou, Yong Yao, Xing Li, Jiu-liang Zhang, Guan-hua Su, Ai-ping Deng

Affiliation(s):  College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China; more

Corresponding email(s):   zjl_ljz@mail.hzau.edu.cn

Key Words:  Gardenblue blueberry (Vaccinium ashei Reade) anthocyanin extracts (GBBAEs), Anti-inflammatory, RAW 264.7, Cyclooxygenase 2 (COX-2), Nuclear factor-κ, B p65 (NF-κ, Bp65)


Wei Xu, Qing Zhou, Yong Yao, Xing Li, Jiu-liang Zhang, Guan-hua Su, Ai-ping Deng. Inhibitory effect of Gardenblue blueberry (Vaccinium ashei Reade) anthocyanin extracts on lipopolysaccharide-stimulated inflammatory response in RAW 264.7 cells[J]. Journal of Zhejiang University Science B, 2016, 17(6): 425-436.

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author="Wei Xu, Qing Zhou, Yong Yao, Xing Li, Jiu-liang Zhang, Guan-hua Su, Ai-ping Deng",
journal="Journal of Zhejiang University Science B",
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pages="425-436",
year="2016",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1500213"
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%T Inhibitory effect of Gardenblue blueberry (Vaccinium ashei Reade) anthocyanin extracts on lipopolysaccharide-stimulated inflammatory response in RAW 264.7 cells
%A Wei Xu
%A Qing Zhou
%A Yong Yao
%A Xing Li
%A Jiu-liang Zhang
%A Guan-hua Su
%A Ai-ping Deng
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A1 - Wei Xu
A1 - Qing Zhou
A1 - Yong Yao
A1 - Xing Li
A1 - Jiu-liang Zhang
A1 - Guan-hua Su
A1 - Ai-ping Deng
J0 - Journal of Zhejiang University Science B
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SP - 425
EP - 436
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PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.B1500213


Abstract: 
Blueberries are a rich source of anthocyanins, which are associated with health benefits contributing to a reduced risk for many diseases. The present study identified the functional gardenblue blueberry (Vaccinium ashei Reade) anthocyanin extracts (GBBAEs) and evaluated their capacity and underlying mechanisms in protecting murine RAW 264.7 cells from lipopolysaccharide (LPS)-stimulated inflammation in vitro. Enzyme-linked immunosorbent assay (ELISA) kit results showed that GBBAEs significantly inhibited the production of nitric oxide (NO), prostaglandin E2 (PGE2), interleukin-6 (IL-6), IL-1β, and interferon-γ (INF-γ). Real-time polymerase chain reaction (PCR) analysis indicated that the mRNA expression levels of IL-6, IL-1β, tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1), and cyclooxygenase 2 (COX-2) were suppressed in LPS-stimulated RAW 264.7 cells. Additionally, Western blot analysis was used to evaluate the relative protein expression levels of COX-2 and nuclear factor-κ;b p65 (NF-κ;bp65). All these results suggested the potential use of GBBAEs as a functional food for the treatment of inflammatory diseases.

蓝莓花青素对RAW 264.7细胞的抗炎活性研究及其机理初探

目的:以园蓝为研究材料,鉴定园蓝花青素提取物(GBBAEs)中的功能性成分的结构,建立脂多糖(LPS)诱导的体外炎症模型,并评价其抗炎作用和初步机制。
创新点:首次探究了蓝莓花青素对建立的LPS诱导体外炎症模型的营养干预作用,并初步探究了发挥抗炎机制的作用通路。
方法:将RAW 264.7细胞分为对照组(不作处理)和实验组(1 µg/ml LPS刺激建模)。实验组进一步分为3个不同浓度组:400 µg/ml GBBAEs组、800 µg/ml GBBAEs组和1200 µg/ml GBBAEs组。用酶联免疫吸附测定(ELISA)试剂盒检测一氧化氮(NO)、前列腺素E2(PGE2)、白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、干扰素γ(INF-γ)等炎症因子的释放量;用实时定量聚合酶链反应(RT-PCR)分析IL-1β、IL-6、TNF-α、环氧合酶-2(COX-2)及单核细胞趋化蛋白-1(MCP-1)的炎症相关基因mRNA的表达水平;用蛋白质印迹法(Western blot法)测定相关炎症蛋白COX-2和NF-κBp65表达水平。
结论:试验结果表明,通过ELISA法测定GBBAEs可以显著性抑制NO、PGE2、IL-1β、IL-6、INF-γ等炎症因子的释放;RT-PCR分析阐明在LPS诱导的单核-巨噬细胞RAW 264.7中,GBBAEs可以显著性抑制IL-1β、IL-6、TNF-α、COX-2及MCP-1的炎症相关基因mRNA的表达水平。此外,Western blot法进一步显示GBBAEs对相关炎症蛋白COX-2和NF-κBp65表达具有明显抑制作用,进一步证实GBBAEs通过NF-κB机制通路来发挥抗炎作用。

关键词:园蓝花青素;抗炎;单核-巨噬细胞RAW 264.7细胞;环氧合酶-2(COX-2);NF-κBp65

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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