目的 對相同母核結(jié)構(gòu)的8種大黃素型蒽醌類化合物開展體外Pig-a基因突變試驗，分析不同大黃素型蒽醌結(jié)構(gòu)與致突變性的關(guān)聯(lián)。方法 L1578Y細(xì)胞分別與系列濃度的大黃素、蘆薈大黃素、大黃素甲醚、大黃酚、大黃酸、羥基大黃素、大黃素-8-O-β-D-葡萄糖苷和蘆薈大黃素-8-O-β-D-葡萄糖苷作用4 h（有S9）或24 h（無S9），給藥24 h后應(yīng)用細(xì)胞計數(shù)板進(jìn)行計數(shù)，計算細(xì)胞相對倍增速率（RPD）評價受試物細(xì)胞毒性；細(xì)胞表達(dá)8 d后經(jīng)APC-anti-CD45和PE-anti-CD90.2標(biāo)定后，使用流式細(xì)胞儀檢測突變細(xì)胞（CD45^＋CD90^－）發(fā)生率。結(jié)果 所有受試物在有或無S9代謝活化條件下所設(shè)濃度組RPD均大于50%，未見明顯細(xì)胞毒性作用，可排除試驗中假陽性結(jié)果。在非S9代謝活化條件下蘆薈大黃素25 μg·mL^-1組Pig-a基因突變率與溶媒對照組比較顯著升高（P<0.001）； S9代謝活化條件下，與溶劑對照組比較，大黃素50 μg·mL^-1組，羥基大黃素6.25、12.5、25 μg·mL^-1組，大黃酚25、50、100 μg·mL^-1組和大黃酸12.5、25、50 μg·mL^-1組Pig-a基因突變率顯著升高（P<0.05、0.01、0.001）。結(jié)論 羥基取代基的多寡及所在位點是蒽醌類化合物致突變性強(qiáng)弱的決定性因素，其體內(nèi)致突變性及致癌性作用仍需進(jìn)行大量體內(nèi)研究證實。

Objective To test eight emodin-type anthraquinone compounds with the same core structure using in vitro Pig-a gene mutation assay, and to analyze the relationship between different emodin-type anthraquinone structures and mutagenicity. Methods L1578Y cells were treated with different concentrations of emodin, aloe-emodin, emodin methyl ether, chrysophanol, rhein, hydroxyemodin, emodin-8-O-β-D-glucoside and aloe-emodin-8-O-β-D-glucoside treatment for 4 h (with S9) or 24 h (without S9). Cell counting plate was used to count 24 h after administration, and the relative cell multiplication rate (RPD) was calculated to evaluate the cytotoxicity of the tested samples. After eight days of expression period, the cells were labelled with APC-anti-CD45 and PE-anti-CD90.2, and the incidence of mutant cells (CD45⁺CD90^-) were detected using flow cytometry. Results The RPD of all tested substances in the concentration group with or without metabolic activation was greater than 50%, and no obvious cytotoxicity was observed, so false positive results in the test could be excluded. The mutation rate of Pig-a gene in aloe emodin 25 μg·mL^-1 group was significantly higher than that in the solvent control group under non-S9 metabolic activation condition (P<0.001). Under S9 metabolic activation condition, compared with solvent control group, the mutation rate of Pig-a gene in emodin 50 μg·mL^-1 group, hydroxyl emodin 6.25, 12.5, 25 μg·mL^-1 groups, chrysophanol 25, 50, 100 μg·mL^-1 groups and rhein 12.5, 25, 50 μg·mL^-1 groups was significantly increased (P<0.05, 0.01, 0.001). Conclusion The number and position of hydroxyl substituents are the decisive factors for the mutagenicity of anthraquinone compounds. However, in vivo mutagenic and carcinogenic effects of emodin-type anthraquinones still need to be confirmed by a large number of in vivo studies.

國家自然科學(xué)基金資助項目（81503347）；國家"十三五""重大新藥創(chuàng)制"專項（2018ZX09201017）