第17卷第1期「論著Antagonistic Effect of Propolis蜂膠乙醇o(jì)n Mutation induced提取物基因oy Thiotepa Adriacin抗突變and Acridine Orange作用的研究WANG Ju-xiang, PEI Shi-geng, men Jin-e,et王菊香丿裘士庚們門金娥3田珂(1. Deparment of Hygien, Hebei engineering College鄭海萍3王延臣2/段曉輝2Handan 056029, China; 2. Derparment of oto Laryngology(1.河北工程學(xué)院衛(wèi)生教研室河北邯鄲the Affiliated Hospital of Hebei Engineering College, Handan 056000056029;2.河北工程學(xué)院附屬醫(yī)院耳鼻喉科China; 3. Experimental Center of the Medical Faculty,河北邯鄲0560003.河北工程學(xué)Hebei Engineering College, Handan 056000, China院醫(yī)學(xué)部中心【摘要】背景與目的探討蜂膠乙醇提取物對噻替哌等誘變劑誘發(fā)的基因突變的抑制作用。材料與方法:用Aπes試驗檢測蜂膠乙醇提取物的誘變性及對噻替哌、阿霉素、吖啶橙誘發(fā)的∏和TAˉ菌株回復(fù)突變后的抑制作用。結(jié)果:在10-1000μ4g/皿蜂膠乙醇提取物對TA灬和'A菌株未岀現(xiàn)回變菌落數(shù)的增高。10~2501g/皿蜂膠濃度出現(xiàn)中等強(qiáng)度或以下強(qiáng)度抑制(抑制率<75%);】000g/皿蜂膠濃度出現(xiàn)高強(qiáng)度回變菌落數(shù)抑制且對試驗所設(shè)3種誘變劑誘發(fā)的基因突變的抑制作用均有劑量-效應(yīng)關(guān)系。結(jié)論:蜂膠對噻替哌等誘變劑所誘發(fā)的不同類型的基因突變均有不同程度的抑制作用。【關(guān)鍵詞】蜂膠;誘變劑;Ames試驗;基因突變中圖分類號:R394.6文獻(xiàn)標(biāo)識碼:A文章編號:1004-616X2005)1-0039-02(ABSTRACT BACKGROUND& AIM: To study the antagonistic effect of propolis on genand other strong mutagens. MATERIAL AND METHODS: Using Ames-test, the antagonistic effect of propopropolis inhibited all the mutations induced by the three kind of mutagens and showed a ,were assessed. RESULTS: Themutation of strains TAlo and TAgg, which induced by thiotepa, adriacin and acridine orangese-enect re10-250 Pg/plate of propolis could inhibit mutations by the middle intensity or below (restrain rate< 75 %). At dose of1 000 Ag/plate of propolis could strongly inhibit mutations induced by thiotepa, adriacin, and acridine orange. There wereno mutant character of the propolis. CONCLUSION: There were antagonistic effect of propolis to gene-mutation induced bydifferent kinds of mutagen. such as adriacinI KEY WORDS propolis: mutagen: Ames test; gene mutation蜂膠是蜜蜂從樹皮的裂縫和植物的幼芽上采集植1材料與方法物的樹脂,加入唾液中的酶和助消化物質(zhì)以及蜂蠟,經(jīng)1.1蜂膠從邯鄲縣某養(yǎng)蜂廠采集新鮮蜂膠,冰咀嚼調(diào)制而成的一種樹脂狀物質(zhì)。研究表明蜂膠中含凍粉碎后以95%乙醇浸泡提取7,h,過濾,濾液于40有多種具有抗菌、抗氧化、抗病毒、調(diào)節(jié)機(jī)體免疫功能及℃減壓揮干二甲基亞砜(DMsO)溶解定容。防癌抗癌功效的生物活性物質(zhì)"·,一些國家作為民間1.2Ames試驗采用平板摻入法。選用TA和用藥有悠久的歷史有文獻(xiàn)報道蜂膠乙醇提取物對環(huán)TAm菌株,菌種引自美國Ames試驗室,經(jīng)中國藥品生磷酰胺等基因突變具有抑制作用,本文用Aπes試驗檢物制品檢定所鑒定菌種特性符合試驗要求。均設(shè)加與不測蜂膠對噻替哌等誘變劑誘發(fā)基因突變的抑制作用。加Ss(由中國藥品生物制品檢定所提供)兩種情況菌株用噻替哌作誘變劑,TA不加S時用阿霉素作誘收稿日期:2004-04-12:修訂日期:2004-08基金項目:邯鄲市科學(xué)技術(shù)研究與發(fā)展項目(1作者簡介:王菊香(1963-),女河北省邯鄲市人副教授研究方向:衛(wèi)生檢驗。Tel:0310-3107382Ⅴol.17No.1變劑,TA加S時用吖啶橙作誘變劑。蜂膠提取物設(shè)42.2蜂膠乙醇提取物抑制誘變試驗當(dāng)蜂膠濃度個濃度組,將45℃融化表層培養(yǎng)基、菌液、陽性物、各為10μg/皿時對吖啶橙引起的TA(+S菌株回復(fù)突變濃度蜂膠提取物、S混合液在滅菌試管中混勻后平鋪到率和噻替哌引起的'A(-Sυ)菌株回復(fù)突變率均無抑制底層培養(yǎng)基上37℃培養(yǎng)48h記數(shù)回變菌落數(shù)并觀察作用;對阿霉素引起的TA(-S)菌株回復(fù)突變率表現(xiàn)菌苔生長情況每組設(shè)3個平行樣重復(fù)兩次。誘變試驗為弱抑制,對噻替哌引起的TA+S)菌株回復(fù)突率表結(jié)果用每皿回變菌落數(shù)和MR值(MR=誘發(fā)回變菌落現(xiàn)為中度抑制。當(dāng)蜂膠濃度為50g/皿時除對噻替哌引數(shù)/自發(fā)回變菌落數(shù))表示MR≥2并有劑量效應(yīng)關(guān)系起的TAx(+S)菌株回復(fù)突率表現(xiàn)為中度抑制外,對其時為誘變陽性。抗誘變試驗結(jié)果用誘變抑制率表示:它陽性物引起的菌株的回復(fù)突變率均表現(xiàn)為弱抑制。當(dāng)抑制率=[(陽性物誘發(fā)回變菌落數(shù)-受試物加陽性物誘發(fā)回變菌蜂膠濃度為250g/皿時,對吖啶橙引起的TA(+S)菌落數(shù))/(陽性物誘發(fā)回變菌落數(shù)-自發(fā)回變菌落數(shù))]×100%。株的回復(fù)突變?nèi)员憩F(xiàn)為弱抑制,對其它陽性物引起的菌抑制率<25%為無抑制,抑制率為25%~50%株的回復(fù)突變率均表現(xiàn)為中度抑制。當(dāng)蜂膠濃度為100(不包括50%)為弱抑制抑制率為50%-75%(不g/m時,4個試驗組均產(chǎn)生強(qiáng)抑制最高一組(噻替哌引包括75%)為中度抑制抑制率≥75%為強(qiáng)抑制。起的TAm(+S組)抑制率達(dá)到978%。直線相關(guān)分析13統(tǒng)計分析采用SAS軟件對測定結(jié)果的劑顯示,四個試驗組均有劑量效應(yīng)關(guān)系(相關(guān)系數(shù)A量-效應(yīng)關(guān)系進(jìn)行直線相關(guān)性分析。S9=0.9684,TAw+S=0.9717,rTA10-S=0.9664,rTA1m+S=0.9655均為P<0.05)。見表22結(jié)果2.1蜂膠乙醇提取物致突變試驗結(jié)果蜂膠濃3討論度在10~1000g/皿范圍內(nèi)未引起突變陽性反應(yīng)(MR許多植物分泌的樹脂類物質(zhì)具有很好的抗菌、防<2)同時也未發(fā)現(xiàn)抑菌現(xiàn)象。見表1。腐、防水、隔熱等作用可以保護(hù)葉、花、果實、幼芽。蜜蜂表1蜂膠對TA和TA1菌株誘發(fā)回變試驗結(jié)果(±x)Table 1 Inducing mutation of propolis on strans TAlon and TAssTAss +SyConcentrationRate ofRate ofg platemutationmutation(×10-2)29.7±2.727.8±31.2434.5±3.31.2157.3±23.31.30161.6±20.71.0146.3±3.032.8±4.2121.0±12.21.00130.8±9.138.0±5.31.2837,8±4,0200.9±16.91.66111.6±11.30.69187.6±17.61.55116.9±14.10.73Adriacin431,0±21.61093.5±52.69.041040.3±50.16.49488.5±38.51.36表2蜂膠抑制TA1和TA誘發(fā)回變試驗結(jié)果(又±s)Table 2 Inhibition of propolis on reverse mutation of strains TAlon and TAysTAoy-SpAx +STAI-S,TAua+SyRate ofMg plateAdriacinAdriacin(×10-221.0±20.3±Propolis280.9±65.5377,9±47.624.0854.3±66,724.6592.9±81.36.5±51.3±61199.8±34.657.6264.6±39.048.6558.6±44.255.0386.5±62.274.31000113.9±11,979,0102,0±27164.8±2195.562,3±22.599,8Thiotep101093.5±52.69.01040.3±50431.0±21.614,51Acridine orangeCompared with group of different propolis concentration, *P<0. 05(下轉(zhuǎn)第44頁)Ⅴol.17No.1化的機(jī)率。因此鎘本身存在的弱遺傳毒性加上它引起「η] Dally h, Hartwig a. Induction and repair inhibition of oxidative的DNA修復(fù)系統(tǒng)功能改變可能是鎘致癌機(jī)制的一個重Dna damage by nickel( II )and cadmium( Il ) in mammalian要方面。cells [J Carcinogenesis, 1997. 18(5): 1 021-1 026[8] Porter DWYakushi參考文獻(xiàn)Escherichia coli(Mutlhuman(MTH1)8-oxo-dGTPases toin vitro inhibition by the carcinogenic metals, nickel( I),co[1] Hartmann A, Speit G Effect of arsenic and cadmium on the(I), cobalt(II ) and cadmium( ll)[J]. Carcinogenesispersistence of mutagen-induced DNA lesions in human cell1997,18(9):1785-1791[J. Environ Mol Mutagen, 1996, 27:98-104[91 Coleman JE. Zinc proteins: enzymes, storage proteins, transcription[21 Hartwig A. Role of DNA repair inhibition in lead- andfactors. and replication proteins[J|. Ann Rew Biochem. 1992admiuminduced genotoxicity: a review[J]. Envir HealthPerspect,194,102(3):45[10 Hartwig A, Asmuss M, Ehleben 1, et al. Interference by toxic13 Fatur T, Lah TT, Filipic M Cadmium inhibits repair of UV-metal ions with DNA repair processes and cell cycle controlmethylmethane-sulfonate and N-methyl-N-nitrosourea-induced DNAmolecular mechanisms[J]. Environ Health Perspect,2002damage in Chinese hamster ovary cells[J. Mutat Res,2003110(5):797-799529(1-2):109-1164]蔡永庭,鄒介智,瞿永華.簡化人全血細(xì)胞DNA非程序合成T11 Hartwig A. Zinc finger proteins as potential targets for toxicnetal ions: differential effects on structure and functionLJI檢測化學(xué)致癌物研究[.腫瘤,1988(2):77-79Antioxid Redox Signal, 2001, 3(4):625-6345]沈芳蘭,張秀珍,李玨聲,等.?；撬釋π∈罂寡趸虳NA[12 Asmuss M. Mullenders LH, Eker A, et al. Differential修復(fù)能力的作用[J」.營養(yǎng)學(xué)報,1996,18(3):258-261effects of toxie metal compounds on the activities of Fpg and[6 Pruski AM, Dixon DR. Effects of cadmium on nuclear integrityXPA, two zinc finger proteins involved in DNA repair[JInd DNA repair efficiells of Mytilus edulis Ieis,2000,21(11):2097-2104Aquat Toxicol,2002,57(3):127-37(上接第40頁)來誘變中間體,或者抑制癌細(xì)胞增殖信號,更重要的是采集樹脂并調(diào)制成蜂膠后,用來加固巢、填補(bǔ)蜂巢縫隙某些抗癌活性可能是直接阻斷接受體或接受體信號旳與空間、涂抹巢房,還用蜂膠密封已被蟄死而不能清除蜂膠抗突變杋制有待進(jìn)一步深λ研究,以便充分開發(fā)這岀巢外的敵害防止其腐爛?？梢姺淠z是一種夭然抗菌種天然抗誘變物質(zhì)的臨床應(yīng)用價值。防腐物質(zhì)。國內(nèi)外大量研究已經(jīng)從世界各地蜂膠中鑒定出參考文獻(xiàn):300多種化合物多種微量元素和維生素2對其中的某1盧志強(qiáng),婁紅祥.植物活性成分與癌癥的化學(xué)預(yù)防[]中草些化合物單項研究證明蜂膠中的多種物質(zhì)具有抗突變藥,2002,33(6:56-565抗癌作用1.5-,根據(jù)蜂膠具有的多種生物活性組分的2郭伽,周立東.蜂膠的化學(xué)成分研究進(jìn)展J.中國養(yǎng)蜂特點,本文用Ames試驗測定了蜂膠的誘變和抗誘變作2000,51(2):17-18用結(jié)果顯示蜂膠無致突變作用能抑制多種誘變劑誘3韓連堂,王志萍,李佩賢,等.蜂膠對環(huán)磷酰胺等四種誘發(fā)的不同類型的基因突變且無論是否經(jīng)肝微粒體酶活變劑誘發(fā)突變的扣制作用[.中國公共衛(wèi)生2001,17(5)化,對陽性物誘發(fā)的突變均有拮抗作用,所以可初步認(rèn)406-4074]周宗燦,傅娟齡,王瑩,等.57種中草藥甲醇提取物體為蜂峰膠具有廣譜的抗誘變作用。外抗突變作用的篩選[J.衛(wèi)生毒理學(xué)雜志,1989,3(4):由于蜂膠抗突變作用的化學(xué)成分非常復(fù)雜,如酚212-214類、香豆素、黃酮類、萜類、維生素AE植物固醇及微量5蔡蕾.食物中非營養(yǎng)成分的生物學(xué)效應(yīng)研究進(jìn)展.國元素等蜂膠抑制基因突變的作用機(jī)制也應(yīng)是一個多方外醫(yī)學(xué)衛(wèi)生學(xué)分冊,1995,22(5):277-281面作用的綜合過程。如蜂膠具有很好的抗氧化作用可6徐厚銓.環(huán)境致突變物和致癌物的抑制劑及其應(yīng)用們J.國以清除氧自由基。其含有的鋅可以摻入DNA聚合酶的外醫(yī)學(xué)胂瘤學(xué)分冊,1989,16(1):26-29合成促進(jìn)DNA損傷的無誤修復(fù),蜂膠中含有的香豆11劉向前,細(xì)胞程序死亡和生物黃酮素,中章草藥,202素、黃酮及酚類可以提高酶系統(tǒng)的活性,促進(jìn)致癌劑滅33(2):190活特別是生物黃酮素可能通過阻止異源物質(zhì)的酶81周土新超氧化物歧化酶及其金屬輔基與惡性腫瘤的關(guān)系J.國外醫(yī)學(xué)衛(wèi)生學(xué)分冊,1995,22(4):228-23