目的 對具有相同母核結(jié)構(gòu)的7種茜素型蒽醌化合物開展體外Pig-a基因突變試驗，分析不同茜素型蒽醌化合物取代基結(jié)構(gòu)與其致突變性的關(guān)聯(lián)。方法 小鼠淋巴瘤細(xì)胞L5178Y（tk^+/--3.7.2.C）分別與系列濃度的茜草素、異茜草素、甲基異茜草素、羥基茜草素、甲基異茜草素-1-甲醚、茜草素-1-甲醚和光澤汀作用4 h（有S9）或24 h（無S9），給藥24 h后應(yīng)用細(xì)胞計數(shù)儀進(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)胞培養(yǎng)8 d后經(jīng)APC-anti-CD45和PE-antiCD90.2抗體孵育后，使用流式細(xì)胞儀檢測細(xì)胞突變（CD45⁺CD90.2^-）率。結(jié)果 所有受試物在有或無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。在無S9代謝活化條件下，光澤汀（16 μg· mL^-1）組、羥基茜草素（10 μg· mL^-1）組、甲基異茜草素-1-甲醚（31.25 μg·mL^-1）組Pig-a基因突變率與溶媒對照組比較顯著升高（P<0.05、0.01、0.001）；在有S9代謝活化條件下，光澤?。?、8 μg· mL^-1）、羥基茜草素（10 μg· mL^-1）、茜草素-1-甲醚（3.75、15.00 μg· mL^-1）、甲基異茜草素（12.5、25.0、50.0、100.0 μ g · mL^-1）、甲基異茜草素-1-甲醚（15、30、60 μ g · mL^-1）、異茜草素（2.50、5.00 μg·mL^-1）組Pig-a基因突變率與溶媒對照組比較顯著升高（P<0.05、0.01、0.001）。結(jié)論 羥基取代基所在位點是茜素型蒽醌化合物致突變性強弱的決定性因素，其體內(nèi)致突變性有待進(jìn)一步確證。

Objective Pig-a gene mutation asay was carried out on seven alizarin type anthraquinone compounds with the same parent nuclear structure in vitro and the relationship between the substituent structure and the mutagenicity were analysed. Methods Alizarin, xanthopurpurin, rubiadin, purpurin, rubiadin -1-methyl, alizarin -1-methyl and lucidin were treated on mouse lymphoma cell L5178Y (tk^+/--3.7.2.C) for 4 hours (-S9) or 24 hours (+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. Cells were cultured for 8 days and APC anti-CD45 antibody/PE-anti-CD90.2 antibody were used to detected mutant cells (CD45⁺CD90^-) by 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. In the absence of S9, the mutant cells (CD45⁺CD90.2^-) were significantly increased by lucidin (16 μg·mL^-1), purpurin (10 μg·mL^-1) and rubiadin-1-methyl ether (31.25 μg·mL^-1) compared with solvent control group. In the presence of S9 metabolic activation, the mutation rate of Pig-a gene in lubutin (4, 8 μg·mL^-1), hydroxyl alizarin (10 μg·mL^-1), alizarin-1-methyl ether (3.75, 15.00 μg·mL^-1), methylisoalizarin (12.5, 25.0, 50.0, 100.0 μg·mL^-1), methylisomadicin-1-methyl ether (15, 30, 60 μg·mL^-1) and isomadicin (2.50, 5.00 μg·mL^-1) groups was significantly higher than that in solvent control group (P<0.05, 0.01, 0.001). Conclusion The location of hydroxyl substituents is a decisive factor in the mutagenicity of alizarin type anthraquinone compounds, while their in vivo mutagenicity need to be further investigated.

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）