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岭南山竹子提取物oblongifolins A的体外抗炎作用及机制研究
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李雪珊 ,覃癸铭 ,石慧颖 ,邹小琴 ,冯 洁 ,钟小斌 (1.广西医科大学第二附属医院药学部,南宁 530007;
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2.南宁市第二人民医院药物临床试验机构办公室,南宁 530031;3.广西医科大学第一附属医院科研部,南宁
530021;4.广西医科大学药学院中药学教研室,南宁 530021)
中图分类号 R965 文献标志码 A 文章编号 1001-0408(2024)10-1209-06
DOI 10.6039/j.issn.1001-0408.2024.10.10
摘 要 目的 研究岭南山竹子提取物oblongifolins A(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(1 μg/mL),地塞米松(DEX)组(10 µmol/L DEX+1 μg/mL LPS),OA低、中、高浓
度组(7.5、15、30 µmol/L OA+1 μg/mL LPS)。除对照组外,其余各组细胞先用LPS刺激1 h后,再与药物混合培养24 h。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-1β mRNA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-κB p65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-1β mRNA表达
水平,磷酸化NF-κB p65(p-NF-κB p65)、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-κB p65 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。
关键词 oblongifolins A;炎症;NF-κB通路;Nrf2通路
Study on in vitro anti-inflammatory effects and mechanisms of oblongifolins A extracted from Garcinia
oblongifolia
LI Xueshan ,QIN Guiming ,SHI Huiying ,ZOU Xiaoqin ,FENG Jie ,ZHONG Xiaobin(1. Dept. of Pharmacy,
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the Second Affiliated Hospital of Guangxi Medical University, Nanning 530007, China;2. Drug Clinical Trial
Facility Office, the Second Nanning People’s Hospital, Nanning 530031, China;3. Dept. of Scientific
Research, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China;4. TCM
Teaching and Research Section, School of Pharmaceutical Sciences, Guangxi Medical University, Nanning
530021, China)
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),
Δ 基金项目 广西自然科学基金项目(No.2023GXNSFAA026233);
p-IκBα, and Kelch-like ECH-associated protein 1 (Keap1)
广西中医药适宜技术开发与推广项目(No.GZSY22-64) were decreased significantly (P<0.05). The contents of IL-4
*第一作者 硕士研究生。研究方向:中药新药研发。E-mail:
1029492357@qq.com and IL-10, protein expressions of IκBα, Nrf2 (except for OA
# 通信作者 主任药师,硕士生导师,硕士。研究方向:中药新药研 low- and medium-concentration groups), HO-1 (except for
发。E-mail:1878125675@qq.com OA low-concentration group) and NQO1 were all increased
中国药房 2024年第35卷第10期 China Pharmacy 2024 Vol. 35 No. 10 · 1209 ·