三种有机磷阻燃剂对BEAS-2B细胞联合毒性的联合指数法测定

Determination of combined toxicity of three organophosphate flame retardants to BEAS-2B cells by combination index

目的:探讨3种有机磷阻燃剂(OPFR)的细胞毒性效应和联合作用类型。方法:将3种OPFR[磷酸三(2-正丁氧乙基)酯(TBOEP)、磷酸三(2-氯丙基)酯(TCIPP)、磷酸三(1,3-二氯-2-丙基)酯(TDCIPP)]以半数抑制浓度(IC_(50))比随机混合,采用CCK-8试剂盒检测其单独和联合对人支气管上皮细胞(BEAS-2B)的毒性作用,并用GraphPad Prism 9对单个化合物及混合物的剂量反应曲线进行非线性拟合。采用CompuSyn软件计算联合指数,判定联合作用的类型及强度。结果:3种OPFR单独对BEAS-2B细胞的毒性大小依次为TDCIPP>TBOEP>TCIPP,其IC_(50)分别是85.8、204.9和758.8μmol/L;联合指数法分析表明TBOEP、TCIPP和TDCIPP在低于IC_(50)的浓度范围内的联合作用类型为协同作用,且随着OPFR浓度的降低,协同作用增强。结论:在低浓度范围内,TBOEP、TCIPP和TDCIPP的联合作用方式为协同作用。

Aim:To test and analyze the cytotoxicity and the combined effects types of three organophosphate flame retardants(OPFR).Methods:Three OPFR[(Tris(2-butoxyethyl)phosphate,TBOEP),(Tris(1-chloro-2-propyl)phosphate,TCIPP),(Tris(1,3-dichloro-2-propyl)phosphate,TDCIPP)]were randomly mixed with the ration of the half maximal inhibitory concentration(IC_(50)).Individual or combined cytotoxicity was assessed using CCK-8 assay and the concentration-response curves(CRC)were fitted by the nonlinear regression model using the GraphPad Prism 9 software.The combination effects of 3 OPFP were assessed using combination index(CI)by the CompuSyn software.Results:The toxicity of the 3 OPFR to BEAS-2B cells decreased as follows:TDCIPP>TBOEP>TCIPP,and the IC_(50)of them were 85.8,204.9 and 758.8μmol/L,respectively.TBOEP,TDCIPP and TCIPP showed synergistic effects by CI analysis when the concentration was below IC_(50).Moreover,the synergistic effects increased with the decrease of concentration.Conclusion:TBOEP,TDCIPP and TCIPP exhibit synergistic effects at low concentration.