【摘要】：目的:開展SD大鼠3天重復(fù)經(jīng)口灌胃給藥毒性實(shí)驗(yàn),聯(lián)合肝、腎和外周血堿性彗星電泳試驗(yàn)和骨髓微核試驗(yàn),并對(duì)遺傳毒性試驗(yàn)常用的溶媒及陽性化合物進(jìn)行檢測(cè)和比較。通過對(duì)比相同實(shí)驗(yàn)條件下開展的以染色體損傷和DNA斷裂為檢測(cè)終點(diǎn)的組合遺傳毒性試驗(yàn)結(jié)果,為給藥和取材時(shí)間點(diǎn)以及陽性劑的選擇提供參考依據(jù)。方法:雄性SD大鼠隨機(jī)分為去離子水組、0.9%氯化鈉注射液組、玉米油組、環(huán)磷酰胺(CPA)10 mg·kg~(-1)組、環(huán)磷酰胺(CPA)40 mg·kg~(-1)組、N-乙基-N-亞硝基脲(ENU)40 mg·kg~(-1)組和甲磺酸乙酯(EMS)200 mg·kg~(-1)組,每組5只。連續(xù)3 d灌胃給藥,實(shí)驗(yàn)期間記錄動(dòng)物臨床癥狀和體重變化。末次給藥后3 h處死大鼠。稱量肝、腎臟器重量,并收集肝、腎及外周血制備單細(xì)胞懸液開展多臟器堿性彗星試驗(yàn)。樣本制片后分別經(jīng)裂解、解旋、電泳、染色等步驟,使用Komet 6.0軟件進(jìn)行圖像分析;取材同時(shí)收集骨髓細(xì)胞制作涂片,計(jì)數(shù)2 000個(gè)嗜多染紅細(xì)胞(PCE)的含微核細(xì)胞發(fā)生率。結(jié)果:1)CPA、ENU和EMS均未對(duì)動(dòng)物整體產(chǎn)生嚴(yán)重的毒性作用。2)只有EMS組的肝細(xì)胞、腎細(xì)胞和淋巴細(xì)胞的刺猬細(xì)胞百分率明顯升高,CPA及ENU對(duì)刺猬細(xì)胞百分率無明顯影響。ENU組在3種組織的彗尾DNA含量百分率及Olive尾矩與溶媒對(duì)照組比較為弱致DNA斷裂作用;而EMS則在3種組織中均顯示了強(qiáng)致DNA斷裂作用。3)CPA作為經(jīng)典的骨髓微核試驗(yàn)陽性劑,其微核結(jié)果顯示了一定的劑量相關(guān)性;ENU的致微核作用相對(duì)較弱。結(jié)論:將多臟器彗星試驗(yàn)與微核試驗(yàn)與3天重復(fù)給藥毒性實(shí)驗(yàn)整合是可行的;實(shí)驗(yàn)流程的標(biāo)準(zhǔn)化、陽性劑的選擇及實(shí)驗(yàn)設(shè)計(jì)等需根據(jù)具體情況來設(shè)計(jì)。
[Abstract]:Aim: to carry out a 3-day repeated oral administration of drugs in SD rats, combined with alkaline comet electrophoresis in liver, kidney and peripheral blood, and micronucleus test in bone marrow, and to detect and compare the common solutes and positive compounds in genotoxicity test. By comparing the results of combinatorial genotoxicity test with chromosome damage and DNA breakage as the end point under the same experimental conditions, this paper provides a reference for the selection of drug administration and sampling time points as well as the selection of positive agents. Methods: male Sprague-Dawley rats were randomly divided into three groups: 0.9% sodium chloride injection group, corn oil group, cyclophosphamide (CPA) 10 mg kg ~ (-1) group, cyclophosphamide (CPA) 40 mg kg ~ (-1) group and (ENU) 40 mg kg ~ (-1) group and (EMS) 200 mg kg ~ (-1) group. The clinical symptoms and body weight of the animals were recorded during the experiment. Rats were killed 3 hours after the last administration. To weigh the weight of liver and kidney, and to collect single cell suspensions from liver, kidney and peripheral blood to carry out alkaline comet assay of multiple viscera. The samples were analyzed by image analysis with Komet 6.0 software, and bone marrow cells were collected to make smears, and the incidence of micronucleus cells in (PCE) of 2000 polychromatic erythrocytes was counted. Results: (1) neither CPA nor EMS had serious toxicity on the whole of animals. 2) there were only hepatocytes in EMS group. The percentage of Hedgehog cells in renal cells and lymphocytes was significantly increased. The percentage of DNA in comet tail and Olive tail moment in three tissues were not significantly affected by CPA and ENU. However, EMS showed strong DNA cleavage (3. 3) CPA as a classic micronucleus positive agent in three tissues. The results of micronucleus showed that the micronucleus effect of CPA was relatively weak in a dose-dependent manner. Conclusion: it is feasible to integrate multiple viscera comet assay with micronucleus test with 3-day repeated administration toxicity test, and the standardization of experimental procedure, the selection of positive agents and the design of experiment should be designed according to the specific conditions.
【作者單位】： 中國食品藥品檢定研究院國家藥物安全評(píng)價(jià)監(jiān)測(cè)中心藥物非臨床安全評(píng)價(jià)研究北京市重點(diǎn)實(shí)驗(yàn)室;中山大學(xué)藥學(xué)院藥物安全評(píng)價(jià)中心;
【基金】：中國食品藥品檢定研究院中青年發(fā)展研究基金課題(2014C1) 國家“重大新藥創(chuàng)制”科技重大專項(xiàng)(2015ZX09501004-002)
【分類號(hào)】：R965

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 文海若;宋捷;陳高峰;齊乃松;李琛;耿興超;王雪;;體外與體內(nèi)肝細(xì)胞微核檢測(cè)方法研究[J];中國新藥雜志;2016年07期

2 劉仕杰;方展強(qiáng);;流式細(xì)胞儀篩選環(huán)磷酰胺誘導(dǎo)骨髓嗜多染紅細(xì)胞微核的技術(shù)方法[J];中國比較醫(yī)學(xué)雜志;2015年03期

3 馬華智;施暢;石富江;孫艷麗;于永生;吳純啟;王全軍;丁日高;;多遺傳學(xué)終點(diǎn)的遺傳毒性試驗(yàn)組合的建立[J];癌變.畸變.突變;2013年06期

4 吳文斌;張超超;湯家銘;;微核試驗(yàn)和彗星試驗(yàn)檢測(cè)雄黃的遺傳毒性[J];中國藥理學(xué)與毒理學(xué)雜志;2012年04期

5 馬愛國,ＡｎｄｒｅｗＲ．Ｃｏｌｌｉｎｓ,ＳｕｓａｎＪ．Ｄｕｔｈｉｅ;不同細(xì)胞 DNA 氧化損傷及自身修復(fù)能力的分析[J];癌變.畸變.突變;1997年03期

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 文海若;毛志慧;陳高峰;宋捷;李琛;楊瑩;王偉凡;王雪;黃芝瑛;;SD大鼠連續(xù)灌胃給藥對(duì)堿性彗星電泳和骨髓微核的影響[J];藥物分析雜志;2017年06期

2 吳露露;汪岱迪;宗衛(wèi)峰;陸益紅;;木糖醇的小鼠骨髓細(xì)胞微核試驗(yàn)[J];海峽藥學(xué);2016年04期

3 馬寶良;魯克慶;;微核試驗(yàn)的改進(jìn)[J];中國比較醫(yī)學(xué)雜志;2016年03期

4 李維;孫開濟(jì);孫玉麗;季昀;周智旋;戴兆來;武振龍;伍國耀;;乳酸菌緩解腸道氧化應(yīng)激研究進(jìn)展[J];動(dòng)物營養(yǎng)學(xué)報(bào);2016年01期

5 韓佳寅;易艷;梁愛華;張宇實(shí);李春英;趙雍;王連嵋;李桂琴;;中藥遺傳毒性研究思路和方法[J];中國中藥雜志;2015年14期

6 張芳芳;洪雅青;朱勇;陳瓊姜;徐娟;陶核;顧劉金;黃雅麗;;無患子皂苷遺傳毒性的研究[J];浙江預(yù)防醫(yī)學(xué);2014年10期

7 惠長野;郭妍;高朝賢;王佃鵬;張文;楊新躍;李智民;;DNA氧化損傷對(duì)人外周血淋巴細(xì)胞微核率的影響[J];國際檢驗(yàn)醫(yī)學(xué)雜志;2014年14期

8 陳穎;李瑩;管雋;王忠輝;賈玉玲;周莉;孫祖越;;標(biāo)準(zhǔn)化Ames波動(dòng)試驗(yàn)最佳條件的優(yōu)選[J];癌變.畸變.突變;2014年03期

9 商慶華;黃盼華;吳文斌;湯家銘;;小鼠淋巴瘤tk基因突變?cè)囼?yàn)檢測(cè)雄黃的致突變性[J];中成藥;2014年05期

10 段長恩;公兵;楊秀濱;;聚丁二酸丁二醇酯浸提液對(duì)人外周血單核細(xì)胞的體外遺傳毒性[J];癌變·畸變·突變;2013年03期

【二級(jí)參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 文海若;淡墨;齊乃松;耿興超;王雪;;多細(xì)胞系胞質(zhì)分裂阻滯微核細(xì)胞組學(xué)試驗(yàn)法的建立與應(yīng)用[J];癌變·畸變·突變;2015年04期

2 Rolf Teschke;Albrecht Wolff;Christian Frenzel;Alexander Schwarzenboeck;Johannes Schulze;Axel Eickhoff;;Drug and herb induced liver injury: Council for International Organizations of Medical Sciences scale for causality assessment[J];World Journal of Hepatology;2014年01期

3 洪麗玲;范賓;黃永焯;劉二剛;王峰;田麗婷;;應(yīng)用胞質(zhì)分裂阻滯微核細(xì)胞組學(xué)法評(píng)價(jià)三溴乙酸的遺傳毒性[J];環(huán)境與職業(yè)醫(yī)學(xué);2013年12期

4 林勃;姜麗平;耿成燕;仲來福;;8-羥基喹啉酮對(duì)HepG2細(xì)胞氧化性DNA損傷的誘導(dǎo)作用(英文)[J];中國藥理學(xué)與毒理學(xué)雜志;2012年02期

5 蔣黎;王宇明;;HepaRG細(xì)胞的生物學(xué)特點(diǎn)及其應(yīng)用[J];肝臟;2010年05期

6 高娜娜;常青;陳莉;李學(xué)彬;楊旭;丁書茂;;氣態(tài)甲醛對(duì)小鼠外周血淋巴細(xì)胞和肝細(xì)胞微核率的影響[J];公共衛(wèi)生與預(yù)防醫(yī)學(xué);2008年03期

7 姚曉峰;仲來福;;人肝癌細(xì)胞系HepG2在遺傳毒物檢測(cè)中的應(yīng)用及其進(jìn)展[J];世界華人消化雜志;2007年02期

8 胡冰冰;楊錄軍;周燕虹;劉勝學(xué);劉晉;曹佳;;小鼠骨髓嗜多染紅細(xì)胞微核單激光流式細(xì)胞儀自動(dòng)化檢測(cè)[J];癌變.畸變.突變;2006年01期

9 孫立平,李德志,劉志謀,楊錄軍,劉晉yN,曹佳;人外周血網(wǎng)織紅細(xì)胞微核的流式細(xì)胞儀自動(dòng)化檢測(cè)[J];癌變.畸變.突變;2004年03期

10 賀爭(zhēng)鳴,李冠民,邢瑞昌;3R理論的形成、發(fā)展及在生命科學(xué)研究中的應(yīng)用[J];實(shí)驗(yàn)動(dòng)物科學(xué)與管理;2000年03期

，

本文編號(hào)：2170716