目的 梳理國(guó)家藥物安全評(píng)價(jià)監(jiān)測(cè)中心聯(lián)合開(kāi)展的大鼠多終點(diǎn)體內(nèi)遺傳毒性試驗(yàn)數(shù)據(jù)，比較大鼠肝彗星試驗(yàn)與骨髓微核試驗(yàn)結(jié)果的一致性和靈敏性。方法 試驗(yàn)分設(shè)陰性物質(zhì)組、作用機(jī)制明確的遺傳毒性陽(yáng)性物質(zhì)組、受試物組，陰性物質(zhì)包括超純水、0.9%氯化鈉注射液、玉米油、0.5%羧甲基纖維素鈉（CMC-Na）、5%蔗糖和聚山梨酯80，給藥體積為10 mL·kg^-1；遺傳毒性陽(yáng)性物質(zhì)包括200 mg·kg^-1甲磺酸乙酯（EMS）、40 mg·kg^-1N-乙基-N-亞硝基脲（ENU）、40 mg·kg^-1環(huán)磷酰胺、75 mg·kg^-1甲基芐肼、800 mg·kg^-1尿烷、75 mg·kg^-1對(duì)氯苯胺、40 mg·kg^-1 1，2-二溴-3-氯丙烷和10 mg·kg^-1秋水仙素；受試物包括100、300、1000 mg·kg^-1大黃素-8-O-β-D-葡萄糖苷，6.5、65.0、650.0 mg·kg^-1單蒽酮和6.5、65.0、650.0 mg·kg^-1大黃素甲醚。分別在實(shí)驗(yàn)0、24、45 hig給藥1次，給藥體積為10 mL·kg^-1。開(kāi)展大鼠肝彗星試驗(yàn)和骨髓微核試驗(yàn)，計(jì)算每只動(dòng)物的肝細(xì)胞刺猬細(xì)胞率和尾DNA百分含量（Tail% DNA），以及每只動(dòng)物的嗜多染紅細(xì)胞（PCE）/總紅細(xì)胞（ERY）比例和嗜多染紅細(xì)胞微核（MNPCE）率。結(jié)果 大鼠肝彗星試驗(yàn)可有效檢出DNA斷裂劑，對(duì)多種烷化劑（甲磺酸乙酯、甲基芐肼和尿烷等）有較好的預(yù)測(cè)性，但對(duì)環(huán)磷酰胺和多倍體誘導(dǎo)劑不靈敏。大黃素-8-O-β-D-葡萄糖苷、單蒽酮和大黃素甲醚骨髓微核試驗(yàn)結(jié)果均為陰性。大黃素-8-O-β-D-葡萄糖苷在1 000 mg·kg^-1劑量下導(dǎo)致的肝Tail% DNA與0.5% CMC-Na組比較顯著升高（P<0.05）；單蒽酮的肝彗星試驗(yàn)結(jié)果為明確陽(yáng)性，劑量為650 mg·kg^-1時(shí)，單蒽酮可導(dǎo)致大鼠肝Tail% DNA顯著升高（P<0.05），且作用存在劑量相關(guān)性；大黃素甲醚的肝彗星試驗(yàn)結(jié)果為陰性。結(jié)論 大鼠肝彗星試驗(yàn)可與骨髓微核試驗(yàn)互補(bǔ)，有效檢出主要作用于肝臟且親電子性較強(qiáng)的遺傳毒性化合物。

Objective The data of multi-endpoint in vivo genotoxicity tests in rats jointly carried out by the National Center for Safety Evaluation of Drugs was analyzed, and the consistency and sensitivity of rat liver comet assay and bone marrow micronucleus test results were compared.Methods The test was divided into negative substance, genotoxic positive substance with clear mechanism of action, and subject group.Negative substances included ultra-pure water, 0.9% sodium chloride injection, corn oil, 0.5% sodium carboxymethyl cellulose(CMC-Na), 5% sucrose and polyssorbide 80, with an administration volume of 10 mL·kg^-1.Genotoxic substances included 200 mg·kg^-1 ethyl mesylate(EMS), 40 mg·kg^-1 N-ethyl-N-nitrourea(ENU), 40 mg·kg^-1 cyclophosphamide, 75 mg·kg^-1 methyl benzyl hydrazine, 800 mg·kg^-1 urane, 75 mg·kg^-1 p-chloroaniline, 40 mg·kg^-1 1, 2-dibromo-3-chloropropane and 10 mg·kg^-1 colchicine.The subjects included 100, 300 and 1 000 mg·kg^-1 emodin-8-O-β-D-glucoside, 6.5, 65.0 and 650.0 mg·kg^-1monanthrone and 6.5, 65.0 and 650.0 mg·kg^-1 emodin methyl ether.At 0, 24 and 45 h of the experiment, the drug was given once ig in a volume of 10 mL·kg^-1.The rat liver comet test and bone marrow micronucleus test were performed.The hepatocyte hedgehog cell rate and Tail% DNA content(Tail% DNA), polychromatic red blood cell(PCE)/total red blood cell(ERY) ratio and polychromatic red blood cell micronucleus(MNPCE) rate of each animal were calculated.Results Rat liver comet assay could effectively detect chemicals inducing DNA breakage and various alkylating agents(ethyl methane sulfonate, methyl benzylhydrazine and urane, etc.), while it was not insensitive to cyclophosphamide and polyploid inducer.The results of bone marrow micronucleus test were negative for emodin-8-O-β-D-glucoside, single anthrone and emodin-methyl ether.The liver% Tail DNA induced by emodin 8-O-β-D-glucoside at 1 000 mg·kg^-1 was significantly increased compared with 0.5% CMC-Na group(P<0.05).The results of liver comet test of single ananthone were clearly positive.At a dose of 650 mg·kg^-1, single ananthone could significantly increase liver%Tail DNA(P<0.05), and there was a dose-dependent effect.The liver comet test for emodin was negative.Conclusion As the second in vivo genotoxicity test recommended by the testing guideline, the rat liver comet assay could complement with the bone marrow micronucleus test and effectively detect genotoxic compounds that mainly act on the liver and are highly electrophilic.

R965

國(guó)家十三五“重大新藥創(chuàng)制”專(zhuān)項(xiàng)課題（2018ZX09201017）；國(guó)家自然科學(xué)基金資助項(xiàng)目（81503347）