3-吲哚甲醇的抗腫瘤作用

華西藥學(xué)雜志2009,23(1):04-047有較好的抑制生長(cháng)作用,而對正常細胞人胚肺成纖具有較好的體外抗腫瘤活性,但粗提物的C∞值偏維細胞MRC-05的細胞毒作用較弱,只在高濃度時(shí)大。經(jīng)過(guò)分離純化得到白花丹醌,實(shí)驗進(jìn)一步研究(0.273gL)顯示較強的抑制生長(cháng)作用了白花丹醌對乳腺癌細胞株(mda-mb-231)的抑表1M法減得自花丹服對md-mb-x及MRC-05的細制作用。眾多抗腫瘤藥物在殺傷腫瘤細胞的同時(shí)胞的體外抑制效應(=5對正常細胞毒性也較大,實(shí)驗以人胚肺成纖維細胞Tabe1 Inhibitory effect of plumbagin on mda-mb-231and(MRC-05)作為正常細胞對照,擬探討白花丹醌對MRC-05 by MTT(n=5)正常細胞的細胞毒作用。結果顯示,白花丹醌對MRC -0Smda-mb-231細胞有較好的抑制生長(cháng)作用,而對inhibitory/% ICso/g L"inhibitory/% ICso/g0.27372.560.014MRC-05細胞毒作用弱。提示白花丹醌的細胞毒作7l.71用可能具有一定的選擇性,其機制值得進(jìn)一步探討0.06970.3969.590.01823魯支[1]劉圓鐘熠高澤文等.白花丹藥材的研究進(jìn)展[門(mén)].華西藥衰2白花丹醌對mda-mb-231克隆形成的抑制作用(x±,=5)學(xué)雜志,2006,21(1):71-73Tabe2 hibitory effect of plumbagin on mda-mb-231bydo[2]劉圓劉超顏曉燕等白花丹不同提取部位體外抗腫瘤及急毒研究[.中國藥理學(xué)通報,2007,23(1):257-258Inhibitory/%1C/gL[3]黃,王建偉張古榮胰島素-5-氟尿啶偶聯(lián)物在體外Control38.67±764的細胞毒性和細胞親和性[J].華西藥學(xué)雜志,2007,22(3):2.31×10-23.67±1.5390.515.13x10-3245-2471.16×10-211.00±7.2171.5[4]曲巍,王立明朱有華等曲古霉素A增強抗腫瘤藥物對膀5.80×10-317.33±4.1655.8胱癌T24細胞的殺傷作用[].中國藥理學(xué)通報,2007,232.90×10-327.00±8.1930.181.45×10-331.33±8.[5]陳奇。中藥藥理研究方法學(xué)[M].第2版.北京:人民衛生出版社,200610152討論收稿日期:2007=0在前期研究中,白花丹的氯仿和石油醚提取物3-吲哚甲醇的抗腫瘤作用羅麗娜,余龍江,郭鳴,敖明章(華中科技大學(xué)生命科學(xué)與技術(shù)學(xué)院湖北武漢430074)摘要:目的探討硫代葡萄糖苷的降解產(chǎn)物3-吲哚甲醇(C)在體內、外的抗癌活性,為十字花科蔬菜的防癌作用以及從蔬菜中篩選藥用成分提供理論依據。方法體內試驗以荷瘤小鼠實(shí)體瘤重、抑瘤率及對脾臟、胸腺指數的影響為指標,評價(jià)C對S移植性肉瘤的抑制作用;長(cháng)期實(shí)驗在飲水中加入化學(xué)致癌劑甲硝基亞硝基胍(MNNG),誘發(fā)小鼠前胃鱗癌及癌前病變,同時(shí)給予不同劑量C進(jìn)行預防觀(guān)察癌前病變和癌變發(fā)生率;體外采用MTT法,觀(guān)察C對Hela細胞增殖的抑制作用。結果LC在體內能明顯抑制實(shí)體瘤重使荷瘤小鼠的脾臟和胸腺指數下降,表現出一定的免疫抑制作用;長(cháng)期試驗能明顯降低MNNG誘發(fā)的癌前病變和癌變發(fā)生率;在體外對Hea細胞有明顯的生長(cháng)抑制作用,且作用隨藥物濃度增加而加大。結論IC具有一定的抗腫瘤活性。關(guān)鍵詞:3-吲哚甲醇;抗腫瘤;硫苷中圖分類(lèi)號:R96文獻標識碼:A文章編號:1006-0103(2008)01-0044-04Experimental study on antitumor activity of indolearoinLUO Li-na, YU Long-jiang, GUO Ming, AO Ming-zhang中國煤化工College of Life Science and Technology, Huazhong University of ScienceCNMHGh作者簡(jiǎn)介:羅麗娜(1981-)女,正攻讀天然產(chǎn)物化學(xué)專(zhuān)業(yè)的碩士學(xué)位。E-mal:lamina0122@163.com通訊作者(Correspondentauthor),E-mil:am25@163.com第1期羅麗娜,等。3-吲哚甲醇的抗腫瘤作用Abstract: OBJECTIVE To investigate the antitumor activity of Indole-3-carbinol (IC), the hydrolytic products of glucosilates, in wioo and in vitro, and to screen medicine materials from diet vegetables and provide theory support for the anticarcinogenicityof crucifers. METHODS The inhibition effect of IC to Suso cells in vivo was evaluated by the tumor weight of the mice bearing tumorand the tumor inhibition rate. The effects on spleen and thymus were also observed. Long term carcinogenesis model was induced byN-methyl-N-nitro-N-nitrosoguanidine( MNNG). The in vitro anticancer effect was studied by MTT assay. RESULTSobtained that HeLa cells were obviously inhibited by the influence of IC in vitro. And there was positive correlation between thetory effect and IC concentration. Furthermore, the IC also had the anticancer activity to the Suso cells of the mice bearing tumor. Thedeclining of spleen and thymus index indicated that IC had the effect of immunosuppressant. IC can reduce the precancerous affectionsand cancer ratio in long term carcinogenesis experiments. coNclusion IC has some effect on inhibition of tumor.Key words: Indole-3-carbinol; Antitumor; GlucosinolateCLC number: R96Document code: AArticle n:1006-0103(2008)01-004硫代葡萄糖苷(硫苷)是一類(lèi)廣泛存在于十字素(深圳華藥南方制藥有限公司,批號:NO60502);花科植物根、莖、果實(shí)中的糖苷類(lèi)混合物,特別是蕓甲硝基亞硝基胍(MNNG,日本東京化成試劑);小鼠苔屬 brassica植物,包括各種卷心菜、椰菜、花椰菜肉瘤細胞株(S3,華中科技大學(xué)同濟醫學(xué)院藥學(xué)和布魯塞爾苗芽。至今已發(fā)現120多種硫苷,在十院);人宮頸癌HeIa細胞(華中科技大學(xué)光電國家字花科植物中已鑒定出27種。根據側鏈基團的重點(diǎn)實(shí)驗室傳代保株)。SF級昆明種小鼠[湖北省氨基酸來(lái)源不同,可將硫苷分為脂肪族、芳香族、吲實(shí)驗研究中心,許可證號:SCXK(鄂)2003-0007哚族硫苷。硫苷本身是一種較為穩定的化合物,但♀♂兼用,體重18-22g。在芥子酶或胃腸道中的細菌酶的催化作用下,會(huì )發(fā)1.2方法與結果生降解并生成多種降解產(chǎn)物2。文獻報道吲哚1.2.1IC對同種移植性腫瘤的作用取接種于昆族硫苷中的3-吲哚甲基硫苷的降解產(chǎn)物3-吲哚明小鼠約7-10d的S1m腫瘤,無(wú)菌條件操作,剔除甲醇(C)可預防癌癥的發(fā)生(圖1)。作者研究了包膜和壞死部分,選取完好的瘤塊,置玻璃勻漿器IC的體內體外的抗腫瘤作用以探討其用于治療內加入適量生理鹽水配制成瘤細胞懸液細胞數癌癥的可能性。為1×107-2×10·ml-,接種于同種異體小鼠前肢/ s-b-D glucosecH。-c腋皮下,每只小鼠注射0.2ml。之后動(dòng)物隨機分組s一 B-D glucose分籠飼養。制成實(shí)體型荷瘤模型。治療組謳供試N-OSO3Indole-3-ylmethyl glucosinolate液,qd;陰性對照組用水;陽(yáng)性對照組用環(huán)磷酰胺(CⅨX),隔日p給藥1次。S1荷瘤小鼠連續給藥10d,停藥次日,脫頸處死動(dòng)物,剝取瘤體組織、脾、胸腺稱(chēng)重,計算腫瘤系數、抑瘤率及臟器系數。采用組間t檢驗及方差分析,應用SPSS12.0統圖1化合物的結構計軟件包進(jìn)行統計學(xué)分析。對S1颱荷瘤小鼠的抑瘤Fig 1 Structures of compounds作用和脾臟、胸腺重量的影響見(jiàn)表1實(shí)驗結果顯示,IC對♂小鼠S1有顯著(zhù)抑制作1實(shí)驗部分用,大小劑量的抑瘤率分別為36.92%34.73%;陽(yáng)1.1儀器、試藥與動(dòng)物性藥物環(huán)磷酰胺可明顯抑制腫瘤的生長(cháng),抑瘤率為二氧化碳培養箱(美國 NuAire);倒置顯微鏡51.5%。環(huán)磷酰胺具有免疫抑制作用,與陰性對照(cOIC);酶標儀(芬蘭 Wallac)。3-吲哚甲醇(LC,組相比,可使脾臟和胸腺系數降低,而大小劑量的Sigma公司,純度99%,批號:122190050);環(huán)磷酰胺C也能使臟器系數降低,其中大劑量C對脾臟的江蘇恒瑞醫藥股份有限公司,批號:06090521);溴抑制作用稍強于環(huán)磷酰胺。由于♀♂個(gè)體差異,在化二甲噻唑二苯四氮唑(MT, Amresco公司分裝);♀荷瘤中國煤化工相差較大,大劑DMEM培養基(Gb公司);新生牛血清(杭州四季量的抑♀小鼠免疫器青生物工程材料有限公司,批號:00201);二甲基亞官具有CNMH腺也有極顯著(zhù)砜(DMSO,天津市大茂化學(xué)試劑廠(chǎng));青霉素鈉(華的抑制作用。北制藥股份有限公司,批號:S0509301);硫酸鏈霉L.22對化學(xué)物質(zhì)誘發(fā)性動(dòng)物腫瘤的作用取華西藥學(xué)雜志第23卷豪1IC對小鼠S1m腫瘤和免疫器官的影響(mgkg1,舞=8Table 1 Influence of IC on 8Sm sarcom mouse and immune organs( mg.kg", m=)aSIa sarcom mouse♀S1 RAnCOmGroupsmg.kg-1 Tc/) Ir/% Oe spleenControl14.4±3.0-12.481±6.821.954±0.4819.3±4.712.173±0.933.568±0.7294±2.6·34.739.861±2.221.881±1.0916.0±4.317.229.958±2.443.160±0.12928.403±1.531.897±04±4.8·41.1311.852±2.06L.841±0.56Compared with control group: P<0. 05,. P<0. 01: Te means tumor coefficient: Ir means inhibition ate Oe means organ coethdear 21.7.0±3.0·51.58.491±1.841.551±0.8511.7±4.1·39.787.791±1.43··1.220±020只小鼠,隨機均分為MNNG組;lC大、中、小三IC的6個(gè)濃度對HeIa細胞作用48h后,與陰性對個(gè)劑量組。取 I g MNNG加1L蒸餾水,避光用磁力照組相比,對腫瘤細胞都有顯著(zhù)的抑制作用,其中攪拌器攪拌約4h,使之溶解成MNNG溶液,置4℃200、250μmol·L劑量的生長(cháng)抑制率分別達到冰箱里避光冷藏。臨用時(shí)配成100μg·m-稀釋6.52%和78.42%;與環(huán)磷酰胺組相比其抑制作用液,置棕色瓶中(外包以鋁箔避光)由小鼠自由飲明顯較強。IC為52.38μmol·L。取,連續12個(gè)月。MNNG組于12個(gè)月后停藥,給自表3IC對HeLa細胞生長(cháng)的影響(48h)(x±,=6)來(lái)水4個(gè)月;C大、中、小劑量組飲水中MNNG用量Tb3 ENect of IC on Hela grow由(48b)(xt,m=6)和時(shí)間與MNNG組相同,分別按100、30、10mgkgig給藥每周6次共16個(gè)月。除實(shí)驗過(guò)程中cmwbInhibitionAmolL-lμnolL-1自然死亡外,其余為有效動(dòng)物,均于16個(gè)月全部處 Control0.236±0.035死。處死動(dòng)物,全面檢查胃腸道,重點(diǎn)觀(guān)察胃。取全cC100.127±0.052·46.26%52.38胃于大彎側剪開(kāi),繪圖或照相后,按0.2~0.4cm寬500.135±0.024·442.81%度全部取材編號,10%甲醛固定,石蠟包埋,間斷連0.152±0.039·35.90%續切片,配E染色。MNNG誘發(fā)小鼠前胃鱗癌的過(guò)0.142±0.03539.84%0.055±0.021·4476.52%程中所出現的增生性病變(角化亢進(jìn)、單純性增生、乳頭狀增生)、癌前病變(乳頭狀瘤、不典型增生)、cx9gm-10173+0.018…2695%癌變(鱗狀細胞癌)與文獻一致。治療組的增生mpared with control group:.P<0. 05,+ P<0. 01; compared with性病變、癌前病變、癌變數均低于MNG組,差異有 CTX甲:△P<0.05,△△P<0.o1顯著(zhù)性(P<0.05)(表2)。表2IC劉MNNG誘發(fā)小鼠胃癌前病變和鱗癌的抑制作用2討論Table 2 Inhibition of carcinogenesis by IC on mice在體內實(shí)驗中,大劑量3-吲哚甲醇(IC)對♀Groups A Dose/mg kg" Hyperplasia/% Precancerous/% Cand%小鼠Sm腫瘤都表現出一定的抑制作用,且大、小Control 28100.042.3·26.9·11.5劑量都具有免疫抑制作用。體外MT實(shí)驗顯示IC2730對HeLa細胞具有明顯的生長(cháng)抑制作用,且抑制作用與藥物濃度呈正相關(guān)。Compared with control group: P<0. 05,.* P<0. 01IC可抑制致癌物與DNA的結合67,除表現出1.2.3MTT比色法測定LC在體外對HeLa細胞的抗機體誘導突變作用、抑制某些酶的活性外,還有抗作用參照文獻,采用培養的對數生長(cháng)期的HeLa激素樣活性及抗細胞增生的作用。細胞,消化、收集細胞,計數,調細胞濃度5x10ml-,接種細胞190在9孔板內。每孔加藥量參考文獻10μ,每種劑量6個(gè)平行孔。藥物作用48h后,每[1] Troyer JK, Stephenson KK, Fahey JW. Analysis of glucosinolates孔加入10μMT,37℃培養箱中培養4h,棄上清,from broccoli and other ouciferous vegetable by hydrophilic inter每孔加150山二甲基亞砜,微量振蕩器振蕩5min,.f 1 1 Chromatogr A,2001,919中國煤化工自動(dòng)酶標讀數儀比色(波長(cháng)570mm)。求出生長(cháng)抑制率(IR)和半數抑制濃度(LCs)。CN MHGohiocyanates: measurementmechanism of action[J]. J Mutation Re采用組間t檢驗及方差分析,應用SPSS12.0統search,2004,555:173-190.計軟件包進(jìn)行統計學(xué)分析(表3)。實(shí)驗結果顯示[3] Nuon- Baudet l, Rabot S,wlJM,da. Actions of the intesti-華西藥學(xué)雜志2008,23(1):047-049Wnal microflora with glucosinolates in rapeseed meal toxicity: first [6] Hudson EA, Howells L, Ball HW. Mechanisms of action of in-evidence of an intestinal lactobacillus possessing a myrosinase likedole-3-carbinol as a chemopreventive agent [J]. Biochem Socactivity in vio[ J].J Sci Food Agric, 1990,52: 547-559Trans,1998,26:S370-371[4] Su Q, Lao ZY, Huo X. Investigation of carcinogenesis induced [7] Kojima T, Tanaka Tby MNNG[ J]. Trancaction of Hengyang Medical college, 1994trial cancer in female donryu rats by indole-3-carbinol[J][5]薛慶善體外培養的原理與技術(shù)[M].北京:科學(xué)出版社,收稿日期:2007-02光源對金絲桃素光動(dòng)力體外抗腫瘤作用的影響王曉利,劉金釧2,張俊松,楊汝德2(1深圳職業(yè)技術(shù)學(xué)院,廣東深圳518055;2.華南理工大學(xué),廣東廣州510641)摘要:目的研究金絲桃素和貫葉金絲桃中金絲桃素提取物對人肝癌細胞株HepG2的體外殺傷效應及光源對其抗腫瘤作用的影響。方法分析金絲桃素及金絲桃素提取物的吸收光譜,用不同波長(cháng)的光源激發(fā)進(jìn)行體外細胞毒試驗,采用顯微鏡觀(guān)察、MTT法分析和DNA電泳等方法評價(jià)金絲桃素及金絲桃素提取物的抗腫瘤效果。結果590m的黃光激發(fā)金絲桃素和金絲桃素提取物對HepG2細胞有顯著(zhù)的體外殺傷效應,生長(cháng)抑制率分別為51.5%、83.1%,其他波長(cháng)光源的作用不顯著(zhù)。結論金絲桃素的光動(dòng)活性依賴(lài)于光源的波長(cháng)受適當光源激發(fā)的金絲桃素及金絲桃素提取物能顯著(zhù)抑制HepC2細胞的生長(cháng),顯示金絲桃素有良好的光動(dòng)力治療肝癌的開(kāi)發(fā)前景。關(guān)鍵詞:金絲桃素;光動(dòng)力治療;肝癌;光波長(cháng)中圖分類(lèi)號:R96文獻標識碼:A文章編號:1006-0103(2008)01-0047-03Effect of light wavelength on photodynamic therapy of hypericin and extract from HypericumL. on HepG2 cancer cell line of human liver in vitroWANG Xiao-li, LIU Jin-chuan, ZHANG Jun-song,YANG Ru-de'(1. Shenzhen Polytechnic, Shenzhen 518055, China; 2.China University of Technology, Guangzhou 510641, China)Abstract: OBJECTTE To investigate the antitumoral effect in mitro and light wavelength effect of hypericin and extract from Hype-ricum perforatum L on HepG2 cancer cell line of human liver. METHODS To detect the absorption spectra of hypericin and ex-tract,use light irradiation of different wavelength for photocytotoxicity test, and assess antitumor effect of hypericin and extract by microscopy, MTT assay and DNA ladder gel electrophoresis. RESULTS The irradiation of 590 nm( yellow)to hypericin and extractad obvious effect on killing HepG2 cell in wuro. The rates of growth inhibition were 51.5 and 83. 1%. CONCLUSION Our studydemonstrates that the photodynamic activity of hypericin and extract depends on light wavelength. Hypericin and extract can obviouslydecrease the growth of HepG2 cancer celL. Hypericin is a powerful naturally photosensitizer and should have a good prospect for furtherKey words: Hypericin; Photodynamic therapy Liver cancer; Light wavelengthCLC number: R96Document code:AArticle id:1006-0103(2008)01-0047-03金絲桃素( hypericin,HY)是貫葉金絲桃 hyper腫瘤方面的報道。文中初步研究了貫葉金絲桃中金cum perforatum L中最具生物活性的物質(zhì),結構為絲桃素提取物受不同光源激發(fā)后對肝癌細胞的作萘駢二蒽酮類(lèi)化合物,可發(fā)出明亮的橘紅色熒光,是用。種良好的光敏劑。研究表明,HY具有抗抑郁、抗病毒(包括Hv)及抗腫瘤等活性。國外在金絲實(shí)驗部分桃素腫瘤光動(dòng)力治疔(PDT)方面的研究開(kāi)發(fā)十分活1.1躍,但未見(jiàn)有關(guān)貫葉金絲桃中金絲桃素提取物在抗YHa中國煤化工度熒光檢測儀CNMHG基金項目:深圳市科技局科技項目(03KB19)作者簡(jiǎn)介:王曉利(1964-),女,碩土,副教授主要從事醫藥生物的科研與教學(xué)工作。E-mal:xiaoli@yahoa.com通訊作者(Correspondentauthor),E-mail:junsongzhangt@yahoo.com.