电子烟烟液体外毒理学评价用细胞筛选研究

Screening of cells for in vitro toxicology test of e-liquid

【目的】目前,国内外尚无电子烟制品体外毒理学评价的统一方法,本研究旨在确定适合于电子烟制品体外毒理学评价用细胞。【方法】以人口腔角质细胞(HOK)、人鼻腔上皮细胞(RPMI 2650)、人支气管上皮细胞(Beas-2b)、人肺成纤维上皮细胞(HPF)、人肺泡上皮细胞(HPAEpiC)为候选细胞,采用细胞毒性法和Annexin V-FITC/PI双染法细胞凋亡试验作为评价方法,综合细胞的耐受性、细胞增殖特性及试验操作性选取适合于电子烟体外毒理学评价用细胞。【结果】细胞毒性和细胞凋亡试验结果均显示,不同的受试细胞对相同的样品测试结果定性结果一致,样品间相对定量趋势一致。候选检测用细胞的细胞倍增时间显示,在24 h的测试周期内,RPMI 2650细胞和HPF细胞的增殖较快。【结论】综合细胞的耐受性、细胞增殖特性及试验操作性,建议选取上呼吸道细胞RPMI 2650和下呼吸道细胞HPF为电子烟体外毒理学测试用细胞。

At present,no standard method has been reported for in vitro toxicology evaluation of e-cigarette.The aim of this study was to find suitable cell lines for in vitro toxicological evaluation of e-cigarettes.Candidate cells were HOK,RPMI 2650,Beas-2b,HPF,HPAEpiC,and MTT.Cytotoxicity method and Annexin V-FITC/PI apoptosis detection method were used as the cell lines selection evaluation method.Cell tolerance,cell proliferation rate and test operability were taken into account to select suitable cell lines for in vitro toxicological evaluation.Diff erent test cell lines were used as test cell,and cytotoxicity and apoptosis test showed that the qualitative results and the relative quantitative trends are consistent.Cell doubling time of candidate cell lines showed that RPMI 2650 and HPF cell lines proliferated faster during the 24h test cycle.It was concluded that the upper respiratory tract RPMI 2650 cell line and lower respiratory tract HPF cell line were suitable as test cell lines for e-cigarette in vitro toxicological evaluation when taking cell tolerance,cell proliferation rate and test operability into account.