發(fā)布時(shí)間：2018-05-10 20:23

  本文選題：升麻 + cycloartane型三萜　； 參考：《浙江大學(xué)》2017年碩士論文

【摘要】：升麻(Cinicifugae Rhizoma)為毛茛科(Ranunculaceae)金蓮花亞科(Subfam.Helleboroideae)升麻族(Cimicifugeae)升麻屬(Cimicifuga L.)多年生草本植物,主要以其地下根莖入藥。在我國(guó)的傳統(tǒng)中醫(yī)理論中升麻有主解百毒,辟瘟疾瘴邪毒蠱和升陽(yáng)舉陷等功效。臨床上主要用于治療風(fēng)熱頭痛,風(fēng)熱表征,齒痛,口瘡,咽喉腫痛,麻疹不透,陽(yáng)毒發(fā)斑;脫肛,子宮脫垂等病癥。利用現(xiàn)代藥理研究方法發(fā)現(xiàn)升麻還具有治療婦女更年期絕經(jīng)的癥狀、抗腫瘤、抗病毒、抗骨質(zhì)疏松、抗炎、抗過(guò)敏、保肝、抑制核苷轉(zhuǎn)運(yùn)、降血脂等功能。本研究首先對(duì)產(chǎn)自中國(guó)東北的升麻(Cimicifuga foetida L.)根莖的95%乙醇提取物的化學(xué)成分和體外藥理活性進(jìn)行了研究。發(fā)現(xiàn)了兩個(gè)吲哚生物堿類(lèi)的新骨架化合物,通過(guò)體外藥理活性篩選發(fā)現(xiàn)它們具有良好的抗腫瘤細(xì)胞活性,因此,我們又進(jìn)一步研究了產(chǎn)自中國(guó)東北的升麻C.foetida(東北)和產(chǎn)自北朝鮮的升麻C.foetida(北朝),利用酸堿萃取的方法富集升麻中的堿性成分。結(jié)合現(xiàn)代色譜手段進(jìn)行系統(tǒng)分離,并對(duì)其光譜波譜數(shù)據(jù)進(jìn)行分析,兩次實(shí)驗(yàn)共分離鑒定了 19個(gè)化合物,分別為:升麻酮堿甲(1);升麻酮堿乙(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為吲哚生物堿類(lèi)的新骨架化合物,化合物11為酚類(lèi)酰胺化合物,化合物10和化合物12為兩個(gè)色原酮類(lèi)化合物,化合物9和19為甾體類(lèi)化合物,其余12個(gè)為cycloartane型三萜類(lèi)化合物。通過(guò)酸堿萃取的方法并沒(méi)有從升麻C.foetida(東北)和升麻C.foetida(北朝)中找到升麻酮堿甲和升麻酮堿乙以及它們的同系物,因此我們又將升麻C.f oetida(東北)萃取后剩下的浸膏通過(guò)大孔樹(shù)脂(D101)富集95%的小極性流分,再通過(guò)Sephadex LH-20、制備薄層TLC等方法進(jìn)行分離,最終成功分離得到了升麻酮堿甲和升麻酮堿乙。隨后我們將升麻酮堿甲和升麻酮堿乙進(jìn)行了體外抗腫瘤活性的研究,結(jié)果表明兩個(gè)化合物對(duì) HL-60、A549、NCI-H1975、Colo-205、A375、MKN7、GSU 這七種腫瘤細(xì)胞系均有顯著的抑制作用,IC50在1.36～21.09μM之間。其中升麻酮堿乙對(duì)人白血病HL-60細(xì)胞的抑制作用尤為明顯,因此,我們進(jìn)一步利用流式細(xì)胞術(shù)和分子蛋白印跡等方法對(duì)升麻酮堿乙抑制HL-60細(xì)胞增殖的作用機(jī)制進(jìn)行了研究,發(fā)現(xiàn)細(xì)胞內(nèi)活化的caspase-3,caspase-8和caspase-9的表達(dá)增多且DNA修復(fù)酶(PARP)的裂解也隨之增多,此外細(xì)胞色素C和Bax蛋白的表達(dá)增多而B(niǎo)c 1-2,Bcl-xL蛋白表達(dá)下降,表明其作用機(jī)制與死亡受體介導(dǎo)的外源性細(xì)胞凋亡通路和線粒體介導(dǎo)的內(nèi)源性細(xì)胞凋亡通路有關(guān)。另外我們將分離得到的Cycloartane型三萜類(lèi)化合物進(jìn)行了體外保肝活性的篩選,但并未發(fā)現(xiàn)明顯的保肝活性。
[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.

【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R284.1

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本文編號(hào)：1870724