岭南山竹子提取物oblongifolinsA的体外抗炎作用及机制研究
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篇名: 岭南山竹子提取物oblongifolinsA的体外抗炎作用及机制研究
TITLE: Study on in vitro anti-inflammatory effects and mechanisms of oblongifolins A extracted from Garcinia oblon-gifolia
摘要: 目的 研究岭南山竹子提取物oblongifolinsA(OA)的体外抗炎作用及机制。方法以RAW264.7细胞为研究对象,将细胞分为对照组(0.5%二甲基亚砜),脂多糖(LPS)组(1μg/mL),地塞米松(DEX)组(10µmol/LDEX+1μg/mLLPS),OA低、中、高浓度组(7.5、15、30µmol/LOA+1μg/mLLPS)。除对照组外,其余各组细胞先用LPS刺激1h后,再与药物混合培养24h。观察各组细胞形态变化,检测各组细胞中一氧化氮(NO)、活性氧(ROS)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-1β、IL-4和IL-10含量,TNF-α、IL-6、IL-1βmRNA表达水平,核因子κB(NF-κB)和核转录因子红系2相关因子2(Nrf2)通路相关蛋白的表达水平以及对照组、LPS组、OA高浓度组细胞中NF-κBp65和Nrf2蛋白的核易位情况。结果与LPS组比较,OA各浓度组纺锤形和不规则细胞逐渐减少,细胞中NO、ROS(OA低浓度组除外)、TNF-α、IL-6、IL-1β含量,TNF-α、IL-6(OA低浓度组除外)、IL-1βmRNA表达水平,磷酸化NF-κBp65(p-NF-κBp65)、磷酸化核因子κB抑制蛋白α(p-IκBα)、Kelch样环氧氯丙烷相关蛋白1(Keap1)蛋白表达水平均显著降低(P<0.05);IL-4、IL-10含量,IκBα、Nrf2(OA低、中浓度组除外)、血红素加氧酶1(HO-1)(OA低浓度组除外)、醌NADH脱氢酶1(NQO1)蛋白表达水平均显著升高(P<0.05);高浓度OA可抑制NF-κBp65蛋白核易位,促进Nrf2蛋白核易位。结论OA能够抑制LPS诱导的RAW264.7细胞炎症,其作用机制可能与抑制NF-κB通路、激活Nrf2通路、减少ROS和炎症因子的过度释放有关。
ABSTRACT: OBJECTIVE To investigate the in vitro anti-inflammatory effects and mechanisms of oblongifolins A (OA) extracted from Garcinia oblongifolia. METHODS RAW264.7 cells were used as the research subject and divided into control group (0.5% DMSO), lipopolysaccharide (LPS) group (1 μg/mL), DEX group (10 µmol/L DEX+1 μg/mL LPS), and low-, medium-, and high-concentration groups of OA (7.5, 15, 30 µmol/L OA+1 μg/mL LPS). Except for the control group, the remaining groups were first stimulated with LPS for 1 hour and then mixed with drugs for 24 hours. The morphological changes of cells were observed in each group. The contents of nitric oxide (NO), reactive oxygen species (ROS), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1β, IL-4 and IL-10 were detected in cells of each group; mRNA expression levels of TNF-α, IL-6 and IL-1β were measured. The expression of key proteins in the nuclear factor κB (NF-κB) and nuclear factor-erythroid 2-related factor 2 (Nrf2) signaling pathways in each group, as well as the nuclear translocation of NF-κB p65 and Nrf2 proteins in control group, LPS group and OA high-concentration group, were detected. RESULTS Compared to the LPS group, the number of spindle-shaped and irregular cells gradually decreased in OA groups, the contents of NO, ROS (except for OA low-concentration group), TNF-α, IL-6 and IL-1β, the mRNA expressions of TNF-α, IL-6 (except for OA low-concentration group) and IL-1β as well as the protein expressions of phosphorylated NF-κB p65 (p-NF-κB p65), p-IκBα, and Kelch-like ECH-associated protein 1 (Keap1) were decreased significantly (P<0.05). The contents of IL-4 and IL-10, protein expressions of IκBα, Nrf2 (except for OA low- and medium-concentration groups), HO-1 (except for OA low-concentration group) and NQO1 were all increased significantly (P<0.05). OA of high concentration could inhibit NF-κB p65 protein nuclear translocation and promote Nrf2 protein nuclear translocation. CONCLUSIONS OA can suppress LPS-induced inflammation in RAW264.7 macrophages. The underlying molecular mechanism likely entails the inhibition of the NF-κB signaling pathway, the activation of the Nrf2 signaling pathway and the reduction of ROS and inflammatory factor release.
期刊: 2024年第35卷第10期
作者: 李雪珊;覃癸铭;石慧颖;邹小琴;冯洁;钟小斌
AUTHORS: LI Xueshan,QIN Guiming,SHI Huiying,ZOU Xiaoqin,FENG Jie,ZHONG Xiaobin
关键字: oblongifolins A;炎症;NF-κB通路;Nrf2通路
KEYWORDS: oblongifolins A; inflammation; NF-κB signaling pathway; Nrf2 signaling pathway
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