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

Journal of Zhejiang University-SCIENCE B, Jun 2016

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-κB p65 (NF-κBp65). All these results suggested the potential use of GBBAEs as a functional food for the treatment of inflammatory diseases.

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

J Zhejiang Univ-Sci B (Biomed & Biotechnol) 1673-1581 Inhibitory effect of Gardenblue blueberry (Vaccinium ashei Reade) anthocyanin extracts on lipopolysaccharide-stimulated Wei XU 0 2 3 5 Qing ZHOU 0 2 4 Yong YAO 0 2 5 Xing LI 0 2 5 Jiu-liang ZHANG 0 2 5 7 Guan-hua SU 0 2 6 Ai-ping DENG 0 2 4 0 Huazhong University of Science and Technology , Wuhan 430014 , China) 1 Project supported by the Fundamental Research Funds for the Central Universities of China (No. 2013PY095), the Clinical Research Project of Health and Family Planning Commission of Wuhan Municipality (No. WX13A05), the Research Project of Wuhan City Central Hospital (No. YQ15A04), and the Grant from Key Laboratory of Biological Targeted Therapy of Hubei Province of China (No. 02.03.2014-10) ORCID: Jiu-liang ZHANG 2 School of Agriculture & Biology, Shanghai Jiao Tong University , Shanghai 200240 , China) 3 Research Center for Food Safety and Nutrition, Key Lab of Urban Agriculture (South), Bor S. Luh Food Safety Research Center 4 Department of Pharmacy, Wuhan City Central Hospital, Tongji Medical College 5 College of Food Science and Technology, Huazhong Agricultural University , Wuhan 430070 , China) 6 Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology , Wuhan 430022 , China) 7 Key Laboratory of Environment Correlative Dietology (Huazhong Agricultural University), Ministry of Education , Wuhan 430070 , China) 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-κB p65 (NF-κBp65). All these results suggested the potential use of GBBAEs as a functional food for the treatment of inflammatory diseases. Gardenblue blueberry (Vaccinium ashei Reade) anthocyanin extracts (GBBAEs); Anti-inflammatory; RAW 264; 7; Cyclooxygenase 2 (COX-2); Nuclear factor-κB p65 (NF-κBp65) http; //dx; doi; org/10; 1631/jzus; B1500213 CLC number; O629; 9 1 Introduction Inflammation response is a complex and crucial process mediated by both several inflammatory cytokines (interferon-γ (INF-γ), interleukin-1β (IL-1β), and IL-6) and related inflammatory mediators such as prostaglandin E2 (PGE2) and nitric oxide (NO), which are produced by cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS), respectively (Kim et al., 2013; Cheng et al., 2014) . COX-2 and nuclear factor-κB (NF-κB) activations are well-known biological markers for inflammatory responses. Inflammatory cells such as monocytes and macrophages can produce a large repertoire of cytokines and participate in the pathogenesis of diseases (Yan et al., 2013). So it is valuable to recognize antiinflammatory mechanisms in order to find treatments and adjuvant treatments of many diseases. Lipopolysaccharide (LPS) is an endotoxin, an integral outer membrane component used to assess Gram-negative bacteria, and triggers the most potent microbial initiators of inflammatory response, including septic shock, fever, and microbial invasion (Dobrovolskaia and Vogel, 2002) . LPS induces NF-κB activation via several signal transduction pathways, all of which inhibit the phosphorylation of NF-κB (inhibitor of NF-κB (IκB)) (Kim et al., 2010) . The LPS-stimulated RAW 264.7 cell model is one of the typical virus-induced inflammation models in vitro. Blueberries are rich in a variety of functional components, such as flavone, organic acid, anthocyanin, superoxide dismutase (SOD), polysaccharides, and mineral elements, contributing to the treatments for many diseases (Almeida et al., 2007) . Although Gardenblue blueberry (Vaccinium ashei Reade) anthocyanin extracts (GBBAEs) are best recognized for their antioxidant properties with their mechanisms being studied thoroughly (Prior et al., 1998; Sun et al., 2012; Li et al., 2013) , recent studies indicated that it is the anti-inflammatory aspect that is responsible for treating cancer and cardiovascular diseases (Johnson et al., 2013; Esposito et al., 2014) . Varieties of polysaccharides, glycosides, and alkaloids have proven to have the best anti-inflamma (...truncated)


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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, Journal of Zhejiang University-SCIENCE B, 2016, pp. 425-436, Volume 17, Issue 6, DOI: 10.1631/jzus.B1500213