[关键词]
[摘要]
目的 对桂枝Cinnamomi Ramulus中非挥发性化学成分及其抗肿瘤、抗炎活性进行研究。方法 运用D101大孔吸附树脂、正反相硅胶、Sephadex LH-20以及半制备HPLC等多种色谱方法进行分离纯化,并根据其波谱数据鉴定出化合物的结构;通过测定化合物对脂多糖(lipopolysaccharide,LPS)诱导小鼠RAW 264.7巨噬细胞释放一氧化氮(NO)的抑制能力,并采用MTT法考察其对人乳腺癌MDA-MB-231细胞的体外抑制能力,进而评价化合物的抗炎和抗肿瘤活性。结果 从桂枝甲醇提取物中共分离得到21个化合物,分别鉴定为表儿茶素(1)、2-(3,4-二羟苯基)乙基-β-D-吡喃葡萄糖苷(2)、3,5-二羟苯基-乙醇-3-O-β-吡喃葡萄糖苷(3)、staphylionoside D(4)、glochidionionoside C(5)、柑橘苷A(6)、dendranthemoside A(7)、euodionoside G(8)、euodionoside F(9)、3,4,5-三甲氧苯基-1-O-β-D-呋喃芹糖-(1'→6')-β-D-吡喃葡萄糖苷(10)、淫羊藿次苷D1(11)、淫羊藿次苷F2(12)、(+)-lyonirenisol-3a-O-β-D-glucopyranoside(13)、l-threo-guaiacylglycerol-8-O-β- glucopyranoside(14)、邻甲氧基肉桂醛(15)、肉桂醛(16)、肉桂酸(17)、4-羟基-3-甲氧基肉桂醛(18)、邻甲氧基苯甲酸(19)、(2R,3R) -5,7-二甲氧基-3',4'-亚甲二氧基黄烷醇(20)、苯甲酸(21)。其中,化合物1、20为黄酮类化合物,2、3、10~12为芳香苷类化合物,4~9为降倍半萜苷类化合物,13、14为苯丙素苷类化合物,15~18为苯丙素类化合物,19、21为苯甲酸类化合物。抗炎、抗肿瘤活性筛选结果表明,化合物15可以有效降低MDA-MB-231细胞的存活能力,其半数抑制浓度(median inhibition concentration,IC50)为(34.11±3.42)μmol/L。化合物15、16、18、20对LPS诱导RAW264.7细胞释放NO具有一定的抑制作用,其IC50分别为(5.27±0.62)、(7.98±2.41)、(7.91±0.81)、(70.34±3.07)μmol/L。结论 化合物2~10、12、14首次从桂枝中分离得到,其中化合物3、5~9、14首次从樟科植物中分离得到,化合物4首次从樟属植物中分离得到;化合物15能有效抑制人乳腺癌MDA-MB-231细胞的生长;化合物15、16、18、20可以显著抑制LPS诱导的RAW264.7细胞的NO释放,具有潜在的抗炎作用;桂枝的化学成分及活性研究为其物质基础研究、质量标准建立以及临床应用提供理论依据。
[Key word]
[Abstract]
Objective To investigate the chemical constituents, antitumor and anti-inflammatory activities of extracts from the non-volatile component of Guizhi (Cinnamomi Ramulus). Methods The compounds were isolated and purified by various chromatographic techniques, including D101, silica gel, Sephadex LH-20 and semi-preparative HPLC, and their structures were identified based on combining spectral analysis with chemical evidence. Their potential anti-tumor and anti-inflammatory effects were evaluated on human breast cell line (MDA-MB-231) and murine macrophage cell line (Raw264.7), respectively. Results A total of 21 compounds were isolated from the methanol extract of C. Ramulus and identified as epicatechin (1), 2-(3,4-dihydroxyphenyl)ethyl-β-D-glucopyranoside (2), 3,5-dihydroxyphenethyl alcohol 3-O-β-glucopyranoside (3), staphylionoside D (4), glochidionionoside C (5), citroside A (6), dendranthemoside A (7), euodionoside G (8), euodionoside F (9), 3,4,5-trimethoxyphenyl-1-O-β-D-apiofuranosyl-(1'→6')glucopyranoside (10), icariside D1 (11), icariside F2 (12), (+)-lyonirenisol-3a-O-β-D-glucopyranoside (13), l-threo-guaiacylglycerol-8-O-β-glucopyranoside (14), O-methoxy cinnamaldehyde (15), cinnamaldehyde (16), cinnamic acid (17), 4-hydroxy-3-methoxycinnamaldehyde (18), O-anisic acid (19), (2R,3R)-5,7- dimethoxy-3',4'-methylenedioxyflavan-3-ol (20), and benzoic acid (21). Compounds 1 and 20 were flavonoids, 2—3 and 10—12 were aromatic glycosides, 4—9 were norsesquiterpenoid glycosides, 13—14 were phenylpropanoid glycosides, 15—18 were phenylpropanoids, and 19 and 21 were phenolic acids. Anti-inflammatory and anti-tumor assays in vitro showed that compound 15 effectively reduced the viability of MDA-MB-231 cells with IC50 of (34.11 ±3.42) μmol/L. Compounds 15—16, 18 and 20 inhibited releases of NO induced by LPS from RAW 264.7 cells with IC50 of (5.27 ±0.62), (7.98 ±2.41), (7.91 ±0.81), and (70.34 ±3.07) μmol/L respectively. Conclusion Compounds 2—10, 12, and 14 are isolated from this plant for the first time. In addition, compounds 3, 5—9, and 14 are found in the Lauraceae for the first time, while compound 4 is isolated from Cinnamomum for the first time. Compound 15 can inhibit the proliferation of human breast cancer cells MDA-MB-231. Compounds 15—16, 18, and 20 can significantly inhibit LPS-induced NO release from RAW264.7, indicating their potential anti-inflammatory effects. The study on the chemical composition, as well as the activity of C. Ramulus may shed new light on its material basis, quality standard establishment and clinical application.
[中图分类号]
[基金项目]
广东省农业科技创新及推广体系建设项目(2021KJ268,2021KJ142,2020KJ142);广东省重点领域研发计划(2020B020221002,2020B1111110003)