小鼠局部淋巴结实验(BrdU-ELISA)在25%氰氟草酯乳油皮肤致敏性检测中的应用
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摘要
目的采用小鼠局部淋巴结实验(BrdU-ELISA)法检测25%氰氟草酯乳油的致敏性。方法 1.选用雌性BALB/C小鼠进行实验,受试样品设为100%、50%、25%,同时设阴性对照组[丙酮:橄榄油(acetone:olive oil,AOO)=4∶1]、阴性对照组(橄榄油)及阳性对照组(25%已基肉桂醛),连续双耳背面染毒3 d,第6天腹腔注射BrdU溶液(5 mg/只),24 h后采集耳后淋巴结称重等并通过BrdU-ELISA实验方法检测淋巴增殖情况。结果各剂量组的小鼠体重与对照组比较,差异均无显著性(P> 0. 05);各剂量组的耳厚度增加和耳重均不超过25%;溶剂和空白对照组的刺激指数SI1和SI2<1. 6,阳性对照组(25%已基肉桂醛)的刺激指数SI1和SI2> 1. 6,实验系统可靠; 25%氰氟草酯乳油低、中、高剂量组的刺激指数SI1和SI2<1. 6,无剂量反应关系。结论 25%氰氟草酯乳油的小鼠局部淋巴结实验结果为阴性,即25%氰氟草酯乳油为无致敏性。
Objective To detect 25% cyhalofop-butyl emulsifiable concentrate( EC) skin sensitization by a local lymph node assay( BrdU-ELISA) in mice. Methods Female BALB/C mice were tested and the samples were set up to100%,50%,and 25%. The negative control group received acetone: olive oil( AOO) = 4 ∶ 1,and the positive control group received 25% basal cinnamaldehyde. Test samples were laid on the dorsum of bilateral mouse ears for three consecutive days,then the mice received an intraperitoneal injection of BrdU( 5 mg/mouse) on the sixth day. After 24 hours,the lymph nodes were removed,weighed,and mashed into a cell suspension. Lymphocyte proliferation was measured by BrdU-ELISA. Results The body weights of female mice in all treated groups were similar to the control group. The increase of ear thickness and ear weight in each dose group was not more than 25%. The stimulus indexes of solvent and blank control groups were SI1 and SI2< 1. 6,and the stimulation indexes of the positive control group( 25% hexyl cinnamaldehyde) were SI1 and SI2> 1. 6. This indicated the test system was reliable. The stimulation indexes of 25%cyhalofop-butyl EC in the low,medium and high dose groups were SI1 and SI2< 1. 6,and there was no dose-response relationship. Conclusions The results of 25% cyhalofop-butyl EC in mouse local lymph node assays were negative,indicating that 25% cyhalofop-butyl does not cause skin sensitization.
Objective To detect 25% cyhalofop-butyl emulsifiable concentrate( EC) skin sensitization by a local lymph node assay( BrdU-ELISA) in mice. Methods Female BALB/C mice were tested and the samples were set up to100%,50%,and 25%. The negative control group received acetone: olive oil( AOO) = 4 ∶ 1,and the positive control group received 25% basal cinnamaldehyde. Test samples were laid on the dorsum of bilateral mouse ears for three consecutive days,then the mice received an intraperitoneal injection of BrdU( 5 mg/mouse) on the sixth day. After 24 hours,the lymph nodes were removed,weighed,and mashed into a cell suspension. Lymphocyte proliferation was measured by BrdU-ELISA. Results The body weights of female mice in all treated groups were similar to the control group. The increase of ear thickness and ear weight in each dose group was not more than 25%. The stimulus indexes of solvent and blank control groups were SI1 and SI2< 1. 6,and the stimulation indexes of the positive control group( 25% hexyl cinnamaldehyde) were SI1 and SI2> 1. 6. This indicated the test system was reliable. The stimulation indexes of 25%cyhalofop-butyl EC in the low,medium and high dose groups were SI1 and SI2< 1. 6,and there was no dose-response relationship. Conclusions The results of 25% cyhalofop-butyl EC in mouse local lymph node assays were negative,indicating that 25% cyhalofop-butyl does not cause skin sensitization.
引文
[1] GB 15670. 9—2017农药登记毒理学试验方法第9部分:皮肤变态反应(致敏)试验[S],2017.
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[3]胡培丽,张露勇,单纯,等. LLNA:BrdU-ELISA改良法在化学物/化妆品皮肤刺激性和致敏性评价中的应用[J].毒理学杂志,2015,29(4):282-285.
[4]张大伟,邢雪,邓乃梅,等.人肝癌细胞培养中CCK-8法与BrdU-ELISA法检测细胞增殖的比较[J].临床普外科电子杂志,2013,1(4):4-7.
[5]胡培丽,张露勇,李波,等.两品系小鼠局部淋巴结试验结果比较[J].中国比较医学杂志,2015,25(5):54-57.
[6]刘珍,刘俊平,万旭英,等.化妆品安全性评价中小鼠局部淋巴结试验方法的改进[J].卫生研究,2009,38(5):585-589.
[7] OECD. OECD guideline for the testing of chemicals, skin sensitisation:local lymph node assay:BrdU-ELISA[S]. Paris:OECD,Adopted 22 July,2010.
[8]阳晓燕,赵康峰,孔建,等.局部淋巴结改良法在化妆品皮肤刺激性和致敏性检测中的应用[J].环境与健康杂志,2013,30(4):312-316.
[9]杨绎雯.我国推行化妆品动物替代实验的可行性研究[J].当代化工研究,2018(2):7-8.
[10] ICCVAM. Test method evaluation report:The murine local lymph node assay:BrdU-ELISA[R]. ICCVAM,2010.
[11]冼静雯,郭煜堂,陈宁,等. BALB/c小鼠LLNA:BrdU-ELISA改良法在15种化学物皮肤致敏性评价中的应用[J].环境与健康杂志,2017,34(5):438-440.
[2]罗涛,王彤敏,李力燕. EdU与BrdU在检测细胞增殖中的特点及应用进展[J].重庆医学,2015,44(32):4581-4583.
[3]胡培丽,张露勇,单纯,等. LLNA:BrdU-ELISA改良法在化学物/化妆品皮肤刺激性和致敏性评价中的应用[J].毒理学杂志,2015,29(4):282-285.
[4]张大伟,邢雪,邓乃梅,等.人肝癌细胞培养中CCK-8法与BrdU-ELISA法检测细胞增殖的比较[J].临床普外科电子杂志,2013,1(4):4-7.
[5]胡培丽,张露勇,李波,等.两品系小鼠局部淋巴结试验结果比较[J].中国比较医学杂志,2015,25(5):54-57.
[6]刘珍,刘俊平,万旭英,等.化妆品安全性评价中小鼠局部淋巴结试验方法的改进[J].卫生研究,2009,38(5):585-589.
[7] OECD. OECD guideline for the testing of chemicals, skin sensitisation:local lymph node assay:BrdU-ELISA[S]. Paris:OECD,Adopted 22 July,2010.
[8]阳晓燕,赵康峰,孔建,等.局部淋巴结改良法在化妆品皮肤刺激性和致敏性检测中的应用[J].环境与健康杂志,2013,30(4):312-316.
[9]杨绎雯.我国推行化妆品动物替代实验的可行性研究[J].当代化工研究,2018(2):7-8.
[10] ICCVAM. Test method evaluation report:The murine local lymph node assay:BrdU-ELISA[R]. ICCVAM,2010.
[11]冼静雯,郭煜堂,陈宁,等. BALB/c小鼠LLNA:BrdU-ELISA改良法在15种化学物皮肤致敏性评价中的应用[J].环境与健康杂志,2017,34(5):438-440.