蛇含委陵菜的木脂素类成分及其细胞毒活性研究

Lignans from Potentilla kleiniana and their cytotoxicity

关键词:  蛇含委陵菜, 化学成分, 分离纯化, 结构鉴定, 木脂素, 细胞毒性, 抗肿瘤活性

Key words:  Potentilla kleiniana, chemical constituents, isolation and purification, structural identification, lignans, cytotoxicity, antitumor activity

摘要:

为了研究蛇含委陵菜(Potentilla kleiniana)的化学成分及其肿瘤细胞毒活性,该研究综合运用D-101大孔树脂、硅胶、Sephadex LH-20、Toyopearl HW-40F及半制备高效液相等现代色谱分离技术对蛇含委陵菜60%乙醇提取物进行分离纯化,根据化合物的理化性质结合核磁共振波谱(NMR)、高分辨质谱(HR-ESI-MS)鉴定化合物的结构,并采用MTT法测定各化合物对人宫颈癌细胞株Hela的细胞毒活性。结果表明:(1)从蛇含委陵菜中分离鉴定了13个木脂素类化合物,分别为(+)-松脂素(1)、(+)-8-羟基松脂素(2)、(+)-丁香脂素(3)、(+)-杜仲树脂酚(4)、(+)-松脂素-4-O-β-D-吡喃葡萄糖苷(5)、(+)-8'-羟基松脂素-4-O-β-D-吡喃葡萄糖苷(6)、(+)-8'-羟基松脂素-4'-O-β-D-吡喃葡萄糖苷(7)、(+)-松脂素-8'-O-β-D-吡喃葡萄糖苷(8)、schilignan F(9)、(+)-松脂素-4, 4'-O-双吡喃葡萄糖苷(10)、(+)-落叶松脂素-4'-O-β-D-吡喃葡萄糖苷(11)、neoolivil-4-O-β-D-glucopyranoside(12)、3,3'-bis [3,4-dihydro-4-hydroxy-6-methoxy-2H-1-benzopyran](13)。其中,化合物1-4、7、8、10、12、13为首次从委陵菜属植物中分离得到,化合物5、6、9、11为首次从蛇含委陵菜中分离得到。(2)细胞毒试验结果显示,化合物1、3、4对Hela细胞具有较好的抑制活性,其半数抑制浓度IC₅₀值分别为(69.94±1.89)、(66.25±2.11)、(59.81±1.73)μmol·L^-1。该研究结果进一步丰富了蛇含委陵菜的化学成分,为抗宫颈癌药物的研发提供物质基础。

Abstract:

Potentilla kleiniana belongs to the family Rosaceae, which distributes in Central Asia, East Asia and Southeast Asia. In China, this plant is mainly found in east, south and southwest provinces. P. kleiniana has been prescribed for the treatment of various diseases in the field of traditional Chinese medicine, such as cough, fever, tuberculosis, mastitis, rheumatoid arthritis. Our previous study found that P. kleiniana had a certain cytotoxicity on tumor cells. The purpose of this paper was to investigate the chemical constituents of P. kleiniana and their cytotoxicity on tumor cells. The 60% ethanol extract of P. kleiniana were isolated by D-101 macroporous adsorptive resins, silica gel, Sephadex LH-20, Toyopearl HW-40F, semi-preparative high performance liquid chromatography and other methods, and their chemical structures were elucidated on the basis of physicochemical properties, NMR and HR-ESI-MS analysis. Meanwhile, all these compounds were evaluated for cytotoxicity against human cervical cancer cell line Hela. The results were as follows:(1)Thirteen lignans were isolated and identified as(+)-pionresinol(1),(+)-8-hydroxypinoresinol(2),(+)-syringaresinol(3),(+)-medioresinol(4),(+)-pionresinol-4-O-β-D-glucopyranoside(5),(+)-8'-hydroxypinoresinol-4-O-β-D-glucopyranoside(6),(+)-8'-hydroxypinoresinol-4'-O-β-D-glucopyranoside(7),(+)-pinoresinol-8'-O-β-D-glucopyranoside(8), schilignan F(9),(+)-pionresinol-4, 4'-O-bisglucopyranoside(10),(+)-lariciresinol-4'-O-β-D-glucopyranoside(11), neoolivil-4-O-β-D-glucopyranoside(12), 3,3'-bis [3,4-dihydro-4-hydroxy-6-methoxy-2H-1-benzopyran](13). Among them, compounds 1-4, 7, 8, 10, 12, 13 were isolated from genus Potentilla for the first time, and compounds 5, 6, 9, 11 were isolated from P. kleiniana for the first time.(2)Cytotoxicity studies showed that compounds 1, 3 and 4 display certain inhibitory activities against Hela cells with IC₅₀ values of(69.94 ± 1.89),(66.25 ± 2.11),(59.81 ± 1.73)μmol·L^-1, respectively. Therefore, the study enriches the chemical constituents of P. kleiniana, and provides a material basis for the development of anti-cervical cancer drugs.