升麻的化学成分研究

发布时间：2018-05-10 20:23

本文选题：升麻 + cycloartane型三萜　；参考：《浙江大学》2017年硕士论文

【摘要】：升麻(Cinicifugae Rhizoma)为毛茛科(Ranunculaceae)金莲花亚科(Subfam.Helleboroideae)升麻族(Cimicifugeae)升麻属(Cimicifuga L.)多年生草本植物,主要以其地下根茎入药。在我国的传统中医理论中升麻有主解百毒,辟瘟疾瘴邪毒蛊和升阳举陷等功效。临床上主要用于治疗风热头痛,风热表征,齿痛,口疮,咽喉肿痛,麻疹不透,阳毒发斑;脱肛,子宫脱垂等病症。利用现代药理研究方法发现升麻还具有治疗妇女更年期绝经的症状、抗肿瘤、抗病毒、抗骨质疏松、抗炎、抗过敏、保肝、抑制核苷转运、降血脂等功能。本研究首先对产自中国东北的升麻(Cimicifuga foetida L.)根茎的95%乙醇提取物的化学成分和体外药理活性进行了研究。发现了两个吲哚生物碱类的新骨架化合物,通过体外药理活性筛选发现它们具有良好的抗肿瘤细胞活性,因此,我们又进一步研究了产自中国东北的升麻C.foetida(东北)和产自北朝鲜的升麻C.foetida(北朝),利用酸碱萃取的方法富集升麻中的碱性成分。结合现代色谱手段进行系统分离,并对其光谱波谱数据进行分析,两次实验共分离鉴定了 19个化合物,分别为:升麻酮碱甲(1);升麻酮碱乙(2);7,8-didehydrocimigenol(3);acerinol(4);24-epi-24-O-acetyl-7,8-didehydrohydroshengmanol(5);cimigenol-3-O-β-D-xylopyranoside(6);24-epi-7,8-didehydrocimigenol-3-O-β-D-xylopyranoside(7);24-epi-24-O-acetyl-7,8-didehydrohydroshengmanol-3-O-β-D-xylopyranoside(8);daucosterol(9);cimifugin(10);cimicifugamide(11);kellol(12);3-xylosyl-24-O-acetylhydroshengmanol-15-glucoside(13);cimigenol(14);24-O-acetyl-25-anhyd roshengmanol-3-β-D-xylopyranoside(15);25-acetylcimigenol-3-O-β-D-xyloside(16);25-anhydrocimigenol-3-O-β-xyloside(17);acetyl-shengmanol-3-O-β-xyloside(18);β-sitosterol(19)。其中化合物1和化合物2为吲哚生物碱类的新骨架化合物,化合物11为酚类酰胺化合物,化合物10和化合物12为两个色原酮类化合物,化合物9和19为甾体类化合物,其余12个为cycloartane型三萜类化合物。通过酸碱萃取的方法并没有从升麻C.foetida(东北)和升麻C.foetida(北朝)中找到升麻酮碱甲和升麻酮碱乙以及它们的同系物,因此我们又将升麻C.f oetida(东北)萃取后剩下的浸膏通过大孔树脂(D101)富集95%的小极性流分,再通过Sephadex LH-20、制备薄层TLC等方法进行分离,最终成功分离得到了升麻酮碱甲和升麻酮碱乙。随后我们将升麻酮碱甲和升麻酮碱乙进行了体外抗肿瘤活性的研究,结果表明两个化合物对 HL-60、A549、NCI-H1975、Colo-205、A375、MKN7、GSU 这七种肿瘤细胞系均有显著的抑制作用,IC50在1.36～21.09μM之间。其中升麻酮碱乙对人白血病HL-60细胞的抑制作用尤为明显,因此,我们进一步利用流式细胞术和分子蛋白印迹等方法对升麻酮碱乙抑制HL-60细胞增殖的作用机制进行了研究,发现细胞内活化的caspase-3,caspase-8和caspase-9的表达增多且DNA修复酶(PARP)的裂解也随之增多,此外细胞色素C和Bax蛋白的表达增多而Bc 1-2,Bcl-xL蛋白表达下降,表明其作用机制与死亡受体介导的外源性细胞凋亡通路和线粒体介导的内源性细胞凋亡通路有关。另外我们将分离得到的Cycloartane型三萜类化合物进行了体外保肝活性的筛选,但并未发现明显的保肝活性。
[Abstract]:Cohosh (Cinicifugae Rhizoma) is a perennial herbaceous plant (Cimicifuga L.) perennial herb (Cimicifuga L.) of the family of Ranunculus (Ranunculaceae) subfamily Subfam.Helleboroideae (Cimicifugeae). It is mainly used as an underground rhizome. It is mainly used in the treatment of wind heat headache, wind heat characterization, toothache, stomatitis, sore throat, sore throat, measles, anus, uterine prolapse and so on. Using modern pharmacological research methods, it is found that cohosh also has the symptoms of menopause for women, anti tumor, anti disease, anti osteoporosis, anti-inflammatory, anti allergic, liver preservation, inhibition of nucleoside transport, In this study, the chemical composition and in vitro pharmacological activity of the 95% ethanol extracts from the rhizomes of Cimicifuga foetida L. from Northeast China were studied. A new skeleton compound of two indole alkaloids was found. The results showed that they had good antitumor cell activity by screening in vitro pharmacological activity. Therefore, we further studied the C.foetida (Northeastern) and the cohosh C.foetida (North Korea), produced from North Korea and North Korea, and enriched the basic components in the cohosh by acid base extraction. The system was separated by modern chromatography, and the spectral data of its spectrum were analyzed, and 19 of the two experiments were separated and identified. Compounds are: cohosh ketone (1); cohosh base B (2); 7,8-didehydrocimigenol (3); acerinol (4); 24-epi-24-O-acetyl-7,8-didehydrohydroshengmanol (5); cimigenol-3-O- beta -D-xylopyranoside (6); 24-epi-7,8-didehydrocimigenol-3-O- beta -D-xylopyranoside (7); 24-epi-24-O-acetyl-7,8-didehydrohydroshengmanol-3-O- beta. D-xylopyranoside (8); daucosterol (9); cimifugin (10); cimicifugamide (11); kellol (12); 3-xylosyl-24-O-acetylhydroshengmanol-15-glucoside (13); cimigenol (14); 24-O-acetyl-25-anhyd roshengmanol-3- beta -D-xylopyranoside (15); 25-acetylcimigenol-3-O- beta -D-xyloside (16); 17; 17; Gmanol-3-O- beta -xyloside (18); beta -sitosterol (19). Compound 1 and compound 2 are new cytoskeleton compounds of indole alkaloids, compound 11 is phenolic amide compound, compound 10 and compound 12 are two chromoone compounds, compounds 9 and 19 are steroids, and the rest 12 are cycloartane three terpenoids. The method of acid and alkali extraction did not find cohosh ketone and cohosh ketone B and their homologues from C.foetida (Northeastern) and cohosh C.foetida (North Korea), so we also enriched 95% small polar flow through macroporous resin (D101) after the extraction of cohosh C.f oetida (northeast), and then prepared by Sephadex LH-20. TLC TLC and other methods were used to separate the cohosh ketone base and cohosh base B. Then we studied the antitumor activity of cohemone and cohosh B in vitro. The results showed that two compounds had significant inhibitory effects on the seven kinds of tumor cell lines, such as HL-60, A549, NCI-H1975, Colo-205, A375, MKN7, GSU. The effect of IC50 is between 1.36 and 21.09 mu M. The inhibitory effect of coimone B on human leukemia HL-60 cells is particularly obvious. Therefore, we have further studied the mechanism of the inhibition of HL-60 cell proliferation by the method of flow cytometry and molecular imprinting, and found the intracellular activation of Caspase-3, caspas. The expression of E-8 and caspase-9 increased and the fragmentation of DNA repair enzyme (PARP) increased. In addition, the expression of cytochrome C and Bax protein increased while the expression of Bc 1-2 and Bcl-xL protein decreased, indicating that the mechanism of action is related to the exogenous apoptosis pathway mediated by death receptor and the mitochondrial mediated endogenous apoptosis pathway. The isolated Cycloartane type three terpenes were screened for hepatoprotective activity in vitro, but no significant hepatoprotective activity was found.

【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R284.1

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