张光甫;刘薇;邢立国;胥维昌;金一和;

• 1:大连理工大学环境学院工业生态与环境工程教育部重点实验室
• 2:中化集团沈阳化工研究院有限公司

摘要(Abstract)：

建立人皮肤模型检测杀菌类农药的腐蚀/刺激性,探讨体外皮肤模型替代农药的动物皮肤安全性评价实验的可行性和影响因素。使用人皮肤成纤维细胞与鼠尾胶原混合培养模拟真皮层,在真皮层上接种人角化细胞,经气液培养形成表皮层。通过细胞活性和细胞因子测定鉴别化合物的腐蚀/刺激性,并比较体外皮肤模型和动物测试结果的一致性。结果表明,皮肤模型对十种农药的腐蚀性测试中,与动物安全评价测试结果完全一致。在刺激性试验中,细胞因子白介素-1a(IL-1a)作为检测终点提高了体外皮肤模型的灵敏性和准确性,人皮肤模型和动物测试结果的一致性达到80%。研究表明,构建的人皮肤模型在对杀菌类农药腐蚀/刺激性测试中初步表现出较好的性能,作为动物替代方法进行农药品安全性评价具有良好的应用前景。

关键词(KeyWords)： 人皮肤模型;腐蚀性;刺激性;杀菌类农药;IL-1a

Abstract:

Keywords:

基金项目(Foundation): 中国博士后科学基金(2012M520646);; 国家重大新药创制专项(2013ZX09302304)

作者(Author): 张光甫;刘薇;邢立国;胥维昌;金一和;

Email:

DOI:

参考文献(References)：

• [1]Frank N,Lehr C M.The human epidermis models EpiSkin,SkinEthic and EpiDerm:An evaluation of morphology and their suitability for testing phototoxicity,irritancy,corrosivity and substance transport[J].Pharmaceutics and Biopharmaceutics,2005,6:167-178
• [2]Kidd D A,Johnson M,Clements J,et al.Development of an in vitro corrosion/irritation prediction assay using the EpiDerm skin model[J].Toxicology in Vitro,2007,21:1292-1297
• [3]Spielmann H,Hoffmann S,Liebsch M,et al.The ECVAM International Validation on in vitro tests for acute skin irritation:Report on the validity of the EMISKIN and EpiDerm assays and on the skin integrity function test[J].Alternatives to Laboratory Animals,2007,35:559-601
• [4]Macfarlane M,Penny J.A tiered approach to the use of alternatives to animals testing for the safety assessment of cosmetics:Skin irritation[J].Regulatory Toxicology and Pharmacology,2009,54:188-196
• [5]Andrea S,Oelgeschlager M.Developmental toxicity testing in the 21stcentury:The sword of Damocles shattered by embryonic stem cell assays?[J].Archives of Toxicology,2011,85:1361-1372
• [6]程树军,秦瑶.体外重建人体皮肤模型刺激试验的验证[J].实验动物与比较医学,2012,32(3):243-246
• [7]邹运动,郑洪艳.基于人体皮肤模型的ECVAM体外皮肤刺激验证研究[J].毒理学杂志,2008,22(3):226-229
• [8]程树军,焦红.体外构建的皮肤模型InspectskinI修复裸鼠全层皮肤缺损[J].中山大学学报,2009,30(2):132-135Cheng S J,Jiao H.Restoration of full-thickness skin defect in nude mice by skin model InspectskinI constructed in vitro[J].Journal of Sun Yat-Sen University(medical sciences),2009,30(2):132-135(in Chinese)
• [9]宋卫国,李宝聚,赵志辉,等.杀菌剂安全风险及解决途径[J].中国蔬菜,2008,1(9):1-4
• [10]Tinois E,Tiollier J,Gaucherand M,Thivolet J,et al.In vitro and post-transplantation differentiation of human keratinocytes grown on the human type IV collagen film of a bilayered dermal substitute[J].Experimental Cell Research,1991,193:310-319
• [11]Cannon C L,Neal P J.New epidermal model for dermal irritancy testing[J].Toxicology in Vitro,1994,8:889-891
• [12]Faller C,Bracher M.Reconstructed skin kit:Reproducibility of cutaneous,irritancy testing,skin pharmacol[J].Skin Physiology,2002,15:74-91
• [13]OECD.OECD guidelines for the testing of chemicals,no.431:In vitro skin corrosion;Human skin model test[S].Paris,France,2004
• [14]OECD.OECD guidelines for the testing of chemicals,no.439:In vitro skin irritation;Human skin model test[S].Paris,France,2010
• [15]Angela C,Jackie Morton.The effects of nickel and chromium on human keratinocytes:Differences in viability,cell associated metal and IL-1a release[J].Toxicology In Vitro,2007,21:809-819
• [16]OECD.OECD guidelines for the testing of chemicals,no.404:Acute dermal irritation/corrosion.Organisation for Economic Co-operation and Development[S]Paris,France,2002
• [17]国家农业部.GB15670-1995农药登记毒理学试验方法[S].国家技术监督局,北京,1995
• [18]程树军,焦红.建立体外重建人体皮肤模型刺激试验的验证标准[J].中山大学学报,2009,5(2):183-189Cheng S J,Jiao H.Validation criterion for reconstruct human skin model to skin irritation[J].Journal of Sun YatSen University,2009,5(2):183-189(in Chinese)
• [19]程树军,潘芳.体外皮肤刺激模型的生物标志研究进展[J].中国比较医学杂志,2010,20(3):64-68Cheng S J,Pan F.Progress on biomarkers of skin irritation model in vitro[J].Chinese Journal of Comparative Medicine,2010,20(3):64-68(in Chinese)
• [20]Abhijit C,Mandip S,et al.In vitro and in vivo comparison of dermal irritancy of jet fuel exposure using EpiDerm cultured human skin and hairless rats[J].Toxicology Letters,2006,167:85-94
• [21]李中良,伍津津.用于替代皮肤刺激试验的组织工程表皮模型的构建研究[J].中国修复重建外科杂志,2011,25(2):129-132Li Z L,Wu J J.Experimental study on constructing a tissue engineered epidermis for replacement of skin irritation in vitro[J].Chinese Journal of Reparative and Reconstructive Surgery,2011,25(2):129-132(in Chinese)
• [22]金岩,吕红兵.人全层组织工程皮肤的研制[J].实用口腔医学杂志,2002,18(2):163-166Jin Y,Lv H B.Development of human tissue engineered skin with full thickness[J].Journal of Practical Stomatology,2002,18(2):163-166(in Chinese)
• [23]Ghalbzouri A,Siamari R.Leiden reconstructed human epidermal model as a tool for the evaluation of the skin corrosion and irritation potential[J].Toxicology in Vitro,2008,22:1311-1320