基于THP-1细胞的皮肤致敏体外检测方法
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广东省科技计划项目(2015A030402005)

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    摘要:

    目的 建立基于人细胞系的替代皮肤致敏动物实验的体外检测方法(h-CLAT),并对化学品、日用化学产品和化妆品植物原料进行皮肤致敏性检测。方法 体外培养人急性单核细胞白血病细胞(THP-1),不同浓度受试物与细胞共孵育24 h,通过流式细胞仪检测细胞表面标志物CD86和CD54活化的差异,并对11种已知皮肤致敏化学物质和9种未知样品进行皮肤致敏性预测。同时对未知样品进行豚鼠局部封闭涂皮法验证。结果 使用h-CLAT方法准确区分了11种参考物质的皮肤致敏性,9种受试样品中,7种样品被判断为阴性皮肤致敏物质,2种植物提取物被确认为皮肤致敏疑似物质。9种样品的预测结果与动物实验一致。结论 h-CLAT体外检测方法可以代替部分动物测试,用于可溶性皮肤致敏物质的预测。

    Abstract:

    Objective To establish an in vitro skin sensitization test, human cell line activation test ( h-CLAT), based on THP-1 cell line (a human acute monocytic leukemia cell line), and to assess the sensitizing potency of plant raw materials of chemical and cosmetic products by this in vitro skin sensitization test. Method THP-1 cells were cultured in vitro and exposed to 11 reference skin sensitization chemicals and 9 samples, by monitoring the cell viability, cell surface marker CD54/CD86 and relative fluorescence intensity of cells surface after the cells was exposures to the substances, and to discover whether there is a positive reaction. At the same time, Buehler test was used to validate the results of samples tested by h-CLAT. Results 11 reference chemicals were distinguished correctly by h-CLAT. Among the 9 samples tested, 7 samples were recognized as negative sensitizer and 2 plant extracted substances were identified as suspicious skin sensitizer. The qualitative classification of the 9 samples by h-CLAT test was consistent with the results obtained by animal test. Conclusions The h-CLAT-in vitro test can be used to replace some animal tests for the prediction of soluble skin sensitizing substances.

    参考文献
    [1] 程树军, 焦红. 实验动物替代方法原理与应用[M]. 北京:科学出版社, 2010:285-304.
    [2] Zurlo J, Hutchinson E. The state of animal welfare in the context of refinement[J]. ALTEX, 2014, 31(1):4-10.
    [3] OECD. The adverse outcome pathway for skin sensitisation initiated by covalent binding to proteins. Part 2:Use of the AOP to develop chemical categories and integrated assessment and testing approaches. Series on testing and assessment No. 168. Paris, 2012. ENV/JM/MONO(2012)/PART2.
    [4] Van Meer PJ, Graham ML, Schuurman HJ. The safety, efficacy and regulatory triangle in drug development:Impact for animal models and the use of animals[J]. Eur J Pharmacol, 2015, 759:3-13.
    [5] Patlewicz G, Kuseva C, Kesova A, et al. Towards AOP application-implementation of an integrated approach to testing and assessment (IATA) into a pipeline tool for skin sensitization[J]. Regul Toxicol Pharmacol, 2014, 69(3):529-545.
    [6] Takenouchi O, Miyazawa M, Saito K, et al. Predictive performance of the human cell line activation test (h-CLAT) for lipophilic chemicals with high octanol-water partition coefficients[J]. J Toxicol Sci, 2013, 38(4):599-609.
    [7] Casati S, Aeby P, Basketter DA, et al. Dendritic cells as a tool for the predictive identification of skin sensitisation hazard[J]. Altern Lab Anim, 2005, 33(1):47-62.
    [8] Nukada Y, Ashikaga T, Miyazawa M, et al. Prediction of skin sensitization potency of chemicals by human Cell Line Activation Test (h-CLAT) and an attempt at classifying skin sensitization potency[J]. Toxicol in Vitro, 2012, 26(7):1150-1160.
    [9] Nukada Y, Ito Y, Miyazawa M. The relationship between CD86 and CD54 protein expression and cytotoxicity following stimulation with contact allergen in THP-1 cells[J]. J Toxicol Sci. 2011, 36(3):313-324.
    [10] Tsujita-Inoue K, Atobe T, Hirota M, et al. In silico risk assessment for skin sensitization using artificial neural network analysis[J], J Toxicol Sci, 2015,40(2):193-209.
    [11] 高向东,吴梧桐.当归及其成分阿魏酸对小鼠免疫功能的影响[J]. 中国生化药物杂志, 1994, 15(2):107-110.
    [12] 曾姣丽, 贺福元, 唐昱, 等. 双黄连注射剂中黄芩苷致敏原性的研究[J]. 中国实验方剂学杂志,2012, 18(1):102-106.
    [13] Tsujita-Inoue KT, Hirota M, Ashikaga T, et al. Skin sensitization risk assessment model using artificial neural network analysis of data from multiple in vitro assays[J]. Toxicol in Vitro, 2014, 28(4):626-639.
    [14] Jaworska J, Harol A, Kern PS, et al. Integrating non-animal test information into an adaptive testing strategy-skin sensitization proof of concept case[J]. ALTEX,2011, 28(3):211-225.
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陈彧,喻欢,秦瑶,程树军,谈伟君.基于THP-1细胞的皮肤致敏体外检测方法[J].中国比较医学杂志,2017,27(4):94~102.

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  • 收稿日期:2016-07-31
  • 在线发布日期: 2017-04-28
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