摘要
目的探讨斑籽木乙酸乙酯萃取部位(BSEA)的体内外抗炎作用及机制。方法将48只雌性小鼠随机分为正常对照组、模型组、阳性对照组及BSEA高、中、低剂量组。阳性对照组给予阿司匹林400 mg/kg灌胃,BSEA高、中、低剂量组分别给予176、88、44 mg/kg浸膏灌胃,模型组给予等体积0.5%羧甲基纤维素钠溶液灌胃,均每天1次,连续灌胃5 d。用角叉菜胶制备小鼠足跖肿胀模型,以足跖肿胀度、足跖肿胀抑制率为观察指标,分析BSEA的体内抗炎作用;对BSEA进行分离纯化,根据波谱数据鉴定化合物结构,观察化合物对脂多糖(LPS)诱导的小鼠RAW264.7巨噬细胞内一氧化氮(NO)生成的抑制作用,评价单体成分的体外抗炎活性;用分子对接技术分析活性成分的抗炎作用机制。结果连续灌胃给药5 d,小鼠未出现异常,表明受试样品在实验所设定的剂量条件下对小鼠的正常生命活动无影响。与模型组比较,阳性对照组、BSEA高剂量组、BSEA中剂量组足肿胀度低(P均<0.05);与阳性对照组比较,BSEA中、低剂量组足肿胀度高(P均<0.05),而高剂量组足肿胀度与阳性对照组比较差异无统计学意义(P>0.05)。阳性对照组及BSEA高、中、低剂量组肿胀抑制率分别为45.56%、35.57%、27.24%、14.19%。从斑籽木BSEA中分离得到4个化合物,分别鉴定为2,3-反式-6,7,4′-三甲氧基黄烷醇(1)、α-亚麻酸(2)、亚麻酸甲酯(3)、甘油亚麻酸酯(4);化合物2对LPS诱导的RAW264.7细胞内NO的生成有一定抑制作用,半数抑制浓度为(22.79±1.48)μmol/L,且其与炎症相关靶标诱导型一氧化氮合酶(iNOS)、过氧化物酶体增长物激活受体α(PPARα)、核转录因子-κB(NF-κB)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)具有较好的亲和力,结合能分别为-7.3、-6.7、-6.4、-6.2、-5.9 kCal/mol。结论BSEA具有较好的体内外抗炎作用,首次从中分离得到化合物1~4,化合物2具有潜在的抗炎作用,其可能通过iNOS、PPARα、NF-κB、IL-1β、TNF-α发挥抗炎作用。
Objective To explore the anti-inflammatory effect and mechanism of the ethyl acetate extracted fraction of baliospermum solanifolium(bsea) in vitro and in vivo. Methods Forty-eight female mice were randomly divided into the normal group, model group, positive control group, and the high-, medium-, and low-dose BSEA groups, respectively. Mice in the positive control group were treated with aspirin(400 mg/kg) by gavage. BSEA(176, 88, and 44 mg/kg, respectively) was given to mice in the high-, medium-, and low-dose groups, and equal volume of 0. 5% sodium carboxymethyl cellulose was given to the mice in the normal group by gavage. All of the above administration groups were given relevant medicine once a day, for consecutive 5 d. Except for the normal group, the acute inflammatory models were established. The carrageenan-induced paw edema in mice was used to explore the anti-inflammatory effects of the BSEA, and the corresponding activity was evaluated via the degree of the foot swelling and the inhibition rate. Chemical constituents were isolated and purified by chromatographic techniques, such as silica gel, MCI, Sephadex LH-20, and semipreparative HPLC, and their structures were identified by analysis of spectral data;Compound 2 was tested for the inhibitory activity against nitric oxide (NO) production in lipopolysaccharide(LPS)-stimulated RAW264. 7 cells and the mechanism of action was investigated by the molecular docking technology. Results After intragastrical administration for consecutive 5 d, no anomaly was observed for the mice, indicating that the tested sample had no effect on the normal vital activity of mice in the corresponding dosage conditions. The swelling degree of feet in the positive control group, and the high-and mediumdose BSEA groups was lower than that in the model group(P<0. 05). Compared with the positive control group, the swelling degree of feet in the medium-and low-dose BSEA groups was higher(P<0. 05). There were no significant differences in the swelling degree of feet between the positive control group and the high-dose BSEA group(P>0. 05). The swelling inhibition rates of the positive control group, and the high-, medium-, and low-dose BSEA groups were 45. 56%, 35. 57%, 27. 24% and 14. 19%, respectively. Four compounds were isolated and identified as 2,3-trans-6,7,4′-trimethoxyflavan-3-ol(1), α-linolenic acid(2), methyl linolenate(3), and 1-linolenoyl glycerol(4). Compound 2 exhibited moderate inhibitory activity against LPS-induced NO production with half-maximal inhibitory concentration(IC50) value of(22. 79±1. 48) μmol/L, and had good affinity to five targets relating to inflammation, inducible nitric oxide synthase(iNOS), peroxisome proliferator-activated receptor α(PPARα), nuclear factor-κB(NF-κB), interleukin-1β(IL-1β), and tumour necrosis factor-α(TNF-α), with the docking binding energy of-7. 3,-6. 7,-6. 4,-6. 2, and-5. 9 kCal/mol, respectively. Conclusion BSEA exhibits anti-inflammatory activity;Compounds 1-4 are isolated from Baliospermum solanifolium for the first time;Compound 2 exhibits potential anti-inflammatory activity, and it may exert the effect by acting on inflammation-related targets such as iNOS, PPARα, NF-κB, IL-1β, and TNF-α.
作者
杨锐
卢光衣
张倩茹
陈帅
何红平
蒋华夷
YANG Rui;LU Guangyi;ZHANG Qianru;CHEN Shuai;HE Hongping;JIANG Huayi(College of Traditional Chinese Medicine,Yunnan University of Chinese Medicine,Kunming 650500,China)
出处
《山东医药》
CAS
2023年第7期42-46,共5页
Shandong Medical Journal
基金
云南省科技厅中医药基础研究联合专项(202101AZ070001-291)
云南省科技人才和平台计划项目(202105AG070012ZD2207)
云南省重大科技专项(202002AA100007)。
关键词
斑籽木
乙酸乙酯萃取部位
体内抗炎作用
体外抗炎作用
Baliospermum solanifolium
ethyl acetate extracted fraction
anti-inflammation in vivo
anti-inflamma‐tion in vitro
