Department of Oriental Medicine Resources and Institute of Korean Herbal Medicine Industry, Mokpo National University, Jeonnam 58554, South Korea.
College of Pharmacy, Mokpo National University, Jeonnam 58554, South Korea.
J Integr Med. 2020 May;18(3):253-264. doi: 10.1016/j.joim.2020.02.001. Epub 2020 Feb 7.
To examine whether Caulerpa okamurae ethanolic extract (COE) could inhibit obesity-mediated inflammation, improve glucose metabolism and increase insulin sensitivity, using in vitro cell models of RAW 264.7 macrophages and 3T3-L1 adipocytes.
We cocultured 3T3-L1 adipocytes in direct contact with lipopolysaccharide-stimulated RAW 264.7 macrophages and induced insulin resistance in 3T3-L1 adipocytes with tumor necrosis factor-α (TNF-α) in the presence or absence of 250 µg/mL of COE. We investigated various markers of inflammation, glucose regulation and insulin sensitivity in these models using Griess reagent to measure nitric oxide (NO) production, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxyglucose to measure glucose uptake, Western blot analysis to quantify protein expression and reverse transcriptase-polymerase chain reaction to evaluate mRNA expression.
We found that COE (250 µg/mL) significantly inhibited the lipopolysaccharide-induced inflammatory response in RAW 264.7 macrophages by downregulating NO production, nitric oxide synthase 2 expression and nuclear translocation of nuclear factor-κB. COE also showed similar anti-inflammatory activity in coculture, along with decreased TNF-α, interleukin-6 and monocyte chemoattractant protein mRNA expression. In addition, COE also improved glucose uptake in coculture by upregulating glucose transporter-4 (GLUT-4) and adiponectin and reducing serine phosphorylation of insulin receptor substrate-1 (IRS1). In the TNF-α-induced insulin resistance model of 3T3-L1 adipocytes, COE significantly improved both basal and insulin-stimulated glucose uptake, accompanied by phosphorylation of IRS1 at tyrosine 632, phospho-5' adenosine monophosphate-activated protein kinase α and glycogen synthase kinase-3β (Ser9) as well as upregulation of GLUT-4.
Together, these findings suggest that COE has potential to treat or prevent obesity-induced metabolic disorders.
使用 RAW 264.7 巨噬细胞和 3T3-L1 脂肪细胞体外细胞模型，研究龟甲龙乙醇提取物（COE）是否能抑制肥胖介导的炎症、改善葡萄糖代谢并增加胰岛素敏感性。
我们将 3T3-L1 脂肪细胞与脂多糖刺激的 RAW 264.7 巨噬细胞共培养，并在存在或不存在 250μg/ml COE 的情况下，用肿瘤坏死因子-α（TNF-α）诱导 3T3-L1 脂肪细胞产生胰岛素抵抗。我们使用 Griess 试剂测量一氧化氮（NO）的产生、2-[[N-(7-硝基苯并-2-氧代-1,3-二唑-4-基)氨基]-2-脱氧-D-葡萄糖测量葡萄糖摄取、Western blot 分析定量蛋白质表达和逆转录-聚合酶链反应评估 mRNA 表达，以此来检测这些模型中的各种炎症、葡萄糖调节和胰岛素敏感性标志物。
我们发现 COE（250μg/ml）通过下调一氧化氮合酶 2 表达和核因子-κB 核易位，显著抑制了脂多糖诱导的 RAW 264.7 巨噬细胞的炎症反应。COE 在共培养物中也表现出类似的抗炎活性，同时降低了 TNF-α、白细胞介素-6 和单核细胞趋化蛋白-1 的 mRNA 表达。此外，COE 还通过上调葡萄糖转运蛋白-4（GLUT-4）和脂联素并减少胰岛素受体底物-1（IRS1）丝氨酸磷酸化，改善了共培养物中的葡萄糖摄取。在 TNF-α诱导的 3T3-L1 脂肪细胞胰岛素抵抗模型中，COE 显著改善了基础和胰岛素刺激的葡萄糖摄取，同时 IRS1 酪氨酸 632、磷酸化 5' 腺苷单磷酸激活蛋白激酶α和糖原合酶激酶-3β（Ser9）磷酸化以及 GLUT-4 上调。
总之，这些发现表明 COE 具有治疗或预防肥胖引起的代谢紊乱的潜力。
