【摘要】：目的:壯藥棵矮瓤系菊科植物馥芳艾納香Blumea aromatica(Wall.)DC.的干燥全草,別名黃藥子、山風(fēng)、香艾、大開(kāi)門(mén)、艾納香等,具有活血止癢、祛風(fēng)消腫的功效,主要用于濕疹,皮膚瘙癢,風(fēng)濕性關(guān)節(jié)炎等癥。壯藥棵矮瓤的化學(xué)成分主要有黃酮類(lèi)、揮發(fā)油類(lèi)、萜類(lèi)、甾體類(lèi)化合物等,目前只有少量文獻(xiàn)對(duì)其化學(xué)成分和藥理活性進(jìn)行研究和報(bào)道。課題組前期對(duì)壯藥棵矮瓤組分進(jìn)行了活性篩選,發(fā)現(xiàn)乙酸乙酯部分具有較好抗肝癌活性,因此,本課題對(duì)其乙酸乙酯部位的化學(xué)成分進(jìn)行了研究,并對(duì)分離出來(lái)的新化合物進(jìn)行了體外抗肝癌活性的初步篩選研究。方法:稱(chēng)取壯藥棵矮瓤藥材50 kg粉粗粉,用500 L 80%乙醇浸泡7 d,過(guò)濾,藥渣再用400 L 80%的乙醇浸提2次,每次7 d,合并3次乙醇提取液,旋蒸回收乙醇,得到3.12 kg壯藥棵矮瓤乙醇提取物浸膏。熱水混懸所得乙醇浸膏后用石油醚和乙酸乙酯依次萃取,回收有機(jī)相,得到602.78 g乙酸乙酯提取部位浸膏。多次反復(fù)采用硅膠色譜法、葡聚糖凝膠色譜法、ODS、Sephadex LH-20、MCI等柱層析以及重結(jié)晶法等現(xiàn)代分離方法和精制純化技術(shù)對(duì)壯藥棵矮瓤乙酸乙酯部位提取物進(jìn)行分離純化,利用~1H-NMR、~(13)C-NMR、MS及文獻(xiàn)參考等方法對(duì)分離提純的單體化合物進(jìn)行結(jié)構(gòu)鑒定。采用MTT方法檢測(cè)新化合物4和14對(duì)肝癌細(xì)胞株(Hep3B和HepG2)的增殖作用�；衔�4和14設(shè)置9個(gè)藥物組(1、5、10、30、70、100、150,200μmol/L)、順鉑組(75μmol/L)以及空白對(duì)照組,分別于給藥24 h、48 h、72 h后,根據(jù)試劑說(shuō)明說(shuō)操作,用酶標(biāo)儀檢測(cè)OD值,并計(jì)算肝癌細(xì)胞增殖抑制率和IC50值。結(jié)果:壯藥棵矮瓤乙酸乙酯部位共分離純化得到16個(gè)單體化合物,分別鑒定為山奈酚⑴、5,7-二羥基-4',6-二甲氧基黃酮醇⑵、木犀草素⑶、(20R)-2α,3α,20-三羥基-16β-乙酰孕甾烷⑷(新)、(20R)-5α-2α,3α,16β,20-四羥基孕甾烷⑸、3,5,7,4’-四羥基-6-甲氧基黃酮醇⑹、β-谷甾醇⑺、β-胡蘿卜苷⑻、3,5,3',4'-O-四甲基槲皮素⑼、3',4',7-三甲氧基槲皮素⑽、艾納香素⑾、沒(méi)食子酸乙酯⑿、沒(méi)食子酸⒀、7-hydroxyspruceanol⒁(新)、芹菜素⒂、香豆酸⒃�；衔�4對(duì)Hep3B和HepG2在48 h的IC50分別為78.15、151.24μmol·L-1,72h分別為55.27μmol·L-1和126.64μmol·L-1。化合物14對(duì)Hep3B和HepG2在48 h的IC50分別為135.80μmol·L-1和223.78μmol·L-1,72 h分別為121.44μmol·L-1和193.82μmol·L-1。結(jié)論:分離鑒定的16個(gè)化合物中,有10個(gè)化合物是首次從棵矮瓤中分離得到,它們分別是5,7-二羥基-4',6-二甲氧基黃酮醇⑵、(20R)-2α,3α,20-三羥基-16β-乙酰孕甾烷⑷(新)、(20R)-5α-2α,3α,16β,20-四羥基孕甾烷⑸、3,5,7,4-四羥基-6-甲氧基黃酮醇⑹、3,5,3',4'-O-四甲基槲皮素⑼、3',4',7-三甲氧基槲皮素⑽、沒(méi)食子酸乙酯⑿、7-hydroxyspruceanol⒁(新)、芹菜素⒂、香豆酸⒃。其中(20R)-2α,3α,20-三羥基-16β-乙酰孕甾烷⑷和7-hydroxyspruceanol⒁為新化合物;在一定濃度梯度下化合物4和14對(duì)肝癌細(xì)胞(Hep3B、HepG2)的增殖均有一定程度的抑制作用,并且呈現(xiàn)濃度依賴(lài)性抑制作用。
[Abstract]:Objective: strong medicine plant dwarf flesh of the Compositae Blumea aromatica (Wall.) DC. dry whole grass, alias xanthate, mountain wind, fragrant Artemisia, big door, aromania and so on, has the effect of activating blood to stop itching, dispelling wind and swelling, mainly used in eczema, skin itching, rheumatic arthritis and other diseases. Oil, terpenoids, steroid compounds and so on, at present only a small amount of literature on the chemical composition and pharmacological activities of the research and report. A preliminary screening study was carried out on the anti hepatoma activity of the isolated new compounds in vitro. Methods: the crude powder of 50 kg powder of strong medicinal herbs was called, and 7 d was soaked with 500 L and 80% ethanol. The residue was filtered by 400 L 80% ethanol for 2 times, 7 d each time, combined with 3 ethanol extracts, and distilled to recover ethanol, and 3.12 kg strong herbs were obtained. The low pulp of 3.12 kg strong medicine was obtained. The extract of ethanol extract was extracted with petroleum ether and ethyl acetate in order to recover organic phase and get the extract of 602.78 g ethyl acetate extract. Many modern separation methods and refining methods, such as silica gel chromatography, dextran gel chromatography, ODS, Sephadex LH-20, MCI and other column chromatography and recrystallization, were repeatedly used. The purification technology was used to isolate and purify the extract of ethyl acetate from the dwarf pulp of Zhuang medicine. The structure identification of the isolated monomers was carried out by ~1H-NMR, ~ (13) C-NMR, MS and literature reference. The proliferation effect of new compound 4 and 14 on the hepatocellular carcinoma cell lines (Hep3B and HepG2) was detected by MTT method. Compound 4 and 14 set up 9 drugs. Group (1,5,10,30,70100150200 mu mol/L), cisplatin group (75 mu mol/L) and blank control group, after administration of 24 h, 48 h, 72 h respectively, according to the reagent description operation, using enzyme labeling instrument to detect the OD value, and calculate the proliferation inhibition rate and IC50 value of liver cancer cells. Results: 16 monomer compounds were isolated and purified from the ethyl acetate fraction of the low flesh of the strong medicine tree, respectively. 5,7- two hydroxyl -4', 6- two methoxy flavonol (2), luteolin 3, (20R) -2 a, 3 alpha, three hydroxyl -16 beta acetosterosterane (New), (20R) -5 alpha -2 a, 3 alpha, 16 beta, 20- four hydroxy gestosterane, 3,5,7,4' - four hydroxy methyl methoxy flavonol, beta carotene, beta carotene, four methyl quercetin 3', 4', 7- trimethoxy quercetin, artemisin, gallic acid, 7-hydroxyspruceanol (New), apigenin, and coumaric acid. Compound 4 to Hep3B and HepG2 in 48 h IC50 are 78.15151.24 micron mol / L-1,72h, respectively, 55.27 Mu mol. C50 was 135.80 Mu mol. L-1 and 223.78 mol. L-1,72 h respectively 121.44 Mu mol. L-1 and 193.82 micron L-1. respectively. Of the 16 compounds identified, 10 compounds were isolated from the low pulp for the first time. They were 5,7- two hydroxyl -4', two methoxy, 3 A and three hydroxy three hydroxy beta acetyl gestopanes. (New) (New) (20R) -5 alpha -2 alpha, 3 alpha, 16 beta, 20- four hydroxyl sterane, 3,5,7,4- four hydroxy -6- methoxy flavonol, 3,5,3', 4'-O- four methyl quercetin, 3', 4', 7- trimethoxy quercetin, ethyl gallate, 7-hydroxyspruceanol (New), apigenin, 3 alpha, three hydroxyl, beta acetyl gestophanus And 7-hydroxyspruceanol as a new compound; under a certain concentration gradient compound 4 and 14 have a certain inhibitory effect on the proliferation of liver cancer cells (Hep3B, HepG2), and present a concentration dependent inhibitory effect.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R29

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 姬志強(qiáng);唐娜娜;張建民;石磊;;山風(fēng)化學(xué)成分研究[J];中國(guó)藥師;2015年12期

2 胡佳;王澤宇;康敏;張兵;鄧峗;;江南紫金牛的化學(xué)成分研究[J];中草藥;2015年14期

3 姜京植;孫倩;許東元;張默函;樸麗花;蔡英蘭;金政;;Forskolin抑制大鼠胃竇環(huán)行平滑肌自發(fā)性收縮運(yùn)動(dòng)[J];生理學(xué)報(bào);2013年02期

4 藍(lán)鳴生;羅超;譚昌恒;陳路;韋松;朱大元;;壯藥山風(fēng)的化學(xué)成分研究[J];中草藥;2012年09期

5 藍(lán)鳴生;羅超;譚昌恒;陳路;韋松;朱大元;;山風(fēng)乙酸乙酯部位的化學(xué)成分研究[J];中藥材;2012年02期

6 楊怡;邵瑋娟;于志文;;高糖條件下forskolin誘導(dǎo)胰島系INS-1細(xì)胞血紅素氧合酶1的表達(dá)[J];中國(guó)現(xiàn)代藥物應(yīng)用;2012年01期

7 張鵬;陸政峰;范志海;李柳炳;沈憶新;;條件培養(yǎng)基、Forskolin與bFGF對(duì)嗅鞘細(xì)胞增殖效應(yīng)的研究[J];臨床醫(yī)學(xué)工程;2011年12期

8 郭海濤;于富敏;李晶超;楊柳;;毛喉萜對(duì)tau蛋白磷酸化及聚集的影響[J];中國(guó)臨床神經(jīng)科學(xué);2011年02期

9 楊博宇;王鋒;王瑋;;牛垂體提取物、堿性成纖維細(xì)胞生長(zhǎng)因子及Forskolin影響嗅鞘細(xì)胞增殖的量效關(guān)系[J];中國(guó)組織工程研究與臨床康復(fù);2010年40期

10 蔣才武;陳毅飛;黃曉燕;盧凱琳;覃青云;孟憲麗;;壯藥牙膏的研制及藥效研究[J];中國(guó)民族民間醫(yī)藥;2010年12期

相關(guān)碩士學(xué)位論文 前2條

1 李芳芳;冰片與抗菌藥物體內(nèi)外聯(lián)合抗菌作用的研究[D];廣東藥學(xué)院;2014年

2 宋愛(ài)華;毛喉鞘蕊花抗腫瘤活性成分及其指紋圖譜研究[D];湖北中醫(yī)學(xué)院;2008年

，

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