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电子烟烟液及其主要化学成分的体外细胞毒性评价

In vitro cytotoxicity evaluation of electronic cigarette liquids and their main chemical components

  • 摘要: 为了解电子烟烟液的细胞毒性以及其主要成分对细胞毒性的影响,通过比对乳酸脱氢酶活性实验(LDH)、溴脱氧尿嘧啶核苷掺入实验(BrdU)、WST-1实验和CCK-8实验检测电子烟烟液对中国仓鼠卵巢细胞(CHO细胞)和人肺癌细胞(A549细胞)的细胞毒性敏感程度,建立了适用于电子烟烟液细胞毒性评价方法,并对16种电子烟烟液进行了分析,通过线性回归分析和Pearson相关分析探讨了电子烟烟液中的烟碱、1,2-丙二醇和丙三醇等与电子烟烟液细胞毒性的相关性。结果表明:①不同实验评价电子烟烟液细胞毒性的IC50值大小为CCK-8 < WST-1 < BrdU < LDH。②电子烟烟液诱导CHO细胞产生的最低毒性效应浓度和半数抑制浓度均低于A549细胞。③16种电子烟烟液的细胞毒性最高相差达5.47倍,但是其细胞毒性与烟液中烟碱、1,2-丙二醇和丙三醇含量(质量分数)的线性回归系数及Pearson相关系数均显著低于线性相关所需水平(P < 0.05)。因此,CHO细胞和CCK-8实验更适合于电子烟烟液的细胞毒性评价,该实验范围内电子烟烟液的细胞毒性与其主要成分(烟碱、丙三醇和1,2-丙二醇)含量之间无明显线性相关性。

     

    Abstract: In order to investigate the cytotoxicity of e-cigarette liquids (e-liquids) and the effects of their main chemical components on the cytotoxicity, a method for evaluating the cytotoxicity of e-liquids was established by comparing responses and detection sensitivities using a range of cytotoxicity assays. These included lactate dehydrogenase assay (LDH), bromodeoxyuridine incorporation assay (BrdU), WST-1 assay, and CCK-8 assay for cytotoxicity of e-liquids to Chinese hamster ovary cells (CHO cells) and human lung cancer cells (A549 cells). A total of 16 e-liquids were analyzed to assess the responses from the market collection. Linear regression and Pearson correlation analyses were used to study the correlation between the cytotoxicity of the e-liquids and the contents of nicotine, 1, 2-propylene glycol and glycerol in the e-liquids. The results showed that:1) The IC50 values of the cytotoxicity of the e-liquids were in the sequence of CCK-8 < WST-1 < BrdU < LDH. 2) The lowest toxic effect concentration and half inhibition concentration of CHO cells induced by the e-liquids were lower than those of A549 cells. 3) Although the difference of the cytotoxicity among the 16 e-liquids was up to 5.47 times, the regression coefficients and Pearson correlation coefficients were significantly lower than those required by the linear correlation (P < 0.05) between the cytotoxicity of the e-liquids and the contents of nicotine, 1, 2-propylene glycol and glycerol. Therefore, CHO cells and CCK-8 assay are more suitable for evaluating the cytotoxicity of e-liquids. Meanwhile, there was no significant linear correlation between the cytotoxicity of the e-liquids and its main components (nicotine, 1, 2-propylene glycol and glycerol) in the experimental range.

     

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