Full Text:   <3030>

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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: 5757

Citations:  Bibtex RefMan EndNote GB/T7714


Jiu-liang Zhang


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


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.

@article{title="Inhibitory effect of Gardenblue blueberry (Vaccinium ashei Reade) anthocyanin extracts on lipopolysaccharide-stimulated inflammatory response in RAW 264.7 cells",
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",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%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
%J Journal of Zhejiang University SCIENCE B
%V 17
%N 6
%P 425-436
%@ 1673-1581
%D 2016
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1500213

T1 - Inhibitory effect of Gardenblue blueberry (Vaccinium ashei Reade) anthocyanin extracts on lipopolysaccharide-stimulated inflammatory response in RAW 264.7 cells
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
VL - 17
IS - 6
SP - 425
EP - 436
%@ 1673-1581
Y1 - 2016
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1500213

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

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


[1]Aggarwal, B.B., 2004. Nuclear factor-κB: the enemy within. Cancer Cell, 6(3):203-208.

[2]Almeida, J., D'Amico, E., Preuss, A., et al., 2007. Characterization of major enzymes and genes involved in flavonoid and proanthocyanidin biosynthesis during fruit development in strawberry (Fragaria × ananassa). Arch. Biochem., 465(1):61-71.

[3]Berliner, J.A., Navab, M., Fogelman, A.M., et al., 1995. Atherosclerosis: basic mechanisms: oxidation, inflammation and genetics. Circulation, 91(9):2488-2496.

[4]Cheng, A.W., Yan, H.Q., Han, C.J., et al., 2014. Polyphenols from blueberries modulate inflammation cytokines in LPS-induced RAW 264.7 macrophages. Int. J. Biol. Macromol., 69:382-387.

[5]Dobrovolskaia, M.A., Vogel, S.N., 2002. Toll receptors, CD14, and macrophage activation and deactivation by LPS. Microbes Infect., 4(9):903-914.

[6]Esposito, D., Chen, A., Grace, M.H., et al., 2014. Inhibitory effects of wild blueberry anthocyanins and other flavonoids on biomarkers of acute and chronic inflammation in vitro. J. Agric. Food Chem., 62(29):7022-7028.

[7]Heinrich, P.C., Behrmann, I., Haan, S., et al., 2003. Principles of interleukin (IL)-6-type cytokine signalling and its regulation. Biochem. J., 374(1):1-20.

[8]Hsu, C.L., Fang, S.C., Liu, C.W., et al., 2013. Inhibitory effects of new varieties of bitter melon on lipopolysaccharide-stimulated inflammatory response in RAW 264.7 cells. J. Funct. Foods, 5(4):1829-1837.

[9]Iqbal, M., Cohen, R.I., Marzouk, K., et al., 2002. Time course of nitric oxide, peroxynitrite and antioxidants in the endotoxemic heart. Crit. Care Med., 30(6):1291-1296.

[10]Johnson, M.H., Mejia, E.G., Fan, J., et al., 2013. Anthocyanins and proanthocyanidins from blueberry-blackberry fermented beverages inhibit markers of inflammation in macrophages and carbohydrate-utilizing enzymes in vitro. Mol. Nutr. Food Res., 57(7):1182-1197.

[11]Kim, K.N., Heo, S.J., Yoon, W.J., et al., 2010. Fucoxanthin inhibits the inflammatory response by suppressing the activation of NF-κB and MAPKs in lipopolysaccharide-induced RAW 264.7 macrophages. Eur. J. Pharmacol., 649(1-3):369-375.

[12]Kim, K.N., Ko, Y.J., Yang, H.M., et al., 2013. Anti-inflammatory effect of essential oil and its constituents from fingered citron (Citrus medica L. var. sarcodactylis) through blocking JNK, ERK and NF-κB signaling pathways in LPS-activated RAW 264.7 cells. Food Chem. Toxicol., 57:126-131.

[13]Korhonen, R., Lahti, A., Kankaanranta, H., et al., 2005. Nitric oxide production and signaling in inflammation. Curr. Drug Target Inflamm. Allergy, 4(4):471-479.

[14]Lau, F.C., Bielinski, D.F., Joseph, J.A., 2007. Inhibitory effects of blueberry extract on the production of inflammatory mediators in lipopolysaccharide-activated BV2 microglia. J. Neurosci. Res., 85(5):1010-1017.

[15]Lee, A.K., Sung, S.H., Kim, Y.C., et al., 2003. Inhibition of lipopolysaccharide-inducible nitric oxide synthase, TNF-α and COX-2 expression by suchinone effects on I-κBα phosphorylation, C/EBP and AP-1 activation. Br. J. Phamacol., 139(1):11-20.

[16]Li, C.Y., Feng, J., Huang, W.Y., et al., 2013. Composition of polyphenols and antioxidant activity of rabbiteye blueberry (Vaccinium ashei) in Nanjing. J. Agric. Food Chem., 61(3):523-531.

[17]Lopes, G., Sousa, C., Silva, L.R., et al., 2012. Can phlorotannins purified extracts constitute a novel pharmacological alternative for microbial infections with associated inflammatory conditions? PLoS ONE, 7(2):e31145.

[18]Mann, J.R., Backlund, M.G., DuBois, R.N., 2005. Mechanisms of disease: inflammatory mediators and cancer prevention. Nat. Clin. Pract. Oncol., 2(4):202-210.

[19]Okamoto, M., Liu, W., Luo, Y., et al., 2010. Constitutively active inflammasome in human melanoma cells mediating autoinflammation via caspase-1 processing and secretion of interleukin-1β. J. Biol. Chem., 285(9):6477-6488.

[20]Prior, R.L., Cao, G., Martin, A., et al., 1998. Antioxidant capacity as influenced by total phenolic and anthocyanin content, maturity and variety of Vaccinium species. J. Agric. Food Chem., 46(7):2686-2693.

[21]Shi, L., Liu, Y.E., Tan, D.H., et al., 2014. Blueberry anthocyanins ameliorate cyclophosphamide-induced liver damage in rats by reducing inflammation and apoptosis. J. Funct. Foods, 11:71-81.

[22]Southan, G.J., Szabo, C., 1996. Selective pharmacological inhibition of distinct nitric oxide synthase isoforms. Biochem. Pharmacol., 51(4):383-394.

[23]Sun, L., Ding, X., Qi, J., et al., 2012. Antioxidant anthocyanins screening through spectrum–effect relationships and DPPH-HPLC-DAD analysis on nine cultivars of introduced rabbiteye blueberry in China. Food Chem., 132(2):759-765.

[24]Wang, L., Nie, Z.K., Zhou, Q., et al., 2014. Antitumor efficacy in H22 tumor bearing mice and immunoregulatory activity on RAW 264.7 macrophages of polysaccharides from Talinum triangulare. Food Funct., 5(9):1979-1980.

[25]Wang, T.Y., Wu, F.H., Jin, Z.G., et al., 2014. Plumbagin inhibits LPS-induced inflammation through the inactivation of the nuclear factor-kappa B and mitogen activated protein kinase signaling pathways in RAW 264.7 cells. Food Chem. Toxicol., 64:177-183.

[26]Wu, X., Beecher, G.R., Holden, J.M., et al., 2006. Concentrations of anthocyanins in common foods in the United States and estimation of normal consumption. J. Agric. Food Chem., 54(11):4069-4075.

[27]Yan, H.Q., Ma, Z.C., Peng, S.A., et al., 2013. Anti-inflammatory effect of auraptene extracted from trifoliate orange (Poncirus trifoliate) on LPS-stimulated RAW 264.7 cells. Inflammation, 36(6):1525-1532.

[28]Yousef, G.G., Brown, A.F., Funakoshi, Y., et al., 2013. Efficient quantification of the health-relevant anthocyanin and phenolic acid profiles in commercial cultivars and breeding selections of blueberries (Vaccinium spp.). J. Agric. Food Chem., 61(20):4806-4815.

[29]Zhang, Z.C., Su, G.H., Luo, C.L., et al., 2015. Effects of anthocyanins from purple sweet potato (Ipomoea batatas L. cultivar Eshu No. 8) on the serum uric acid level and xanthine oxidase activity in hyperuricemic mice. Food Funct., 6(9):3045-3055.

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