氯丙嗪和噻洛芬酸在3种光毒性测试中致p38和Erk1/2表达变化研究
Chlorpromazine and Tiaprofenic Acid-induced p38 and Erk1/2 Expression Change in Three Phototoxicity Test Methods
吴晓星 1陈舒怀 2刘彦娟 3桑晶2
作者信息
- 1. 浙江大学医学院附属第二医院基础和转化研究中心,杭州 310003
- 2. 浙江省食品药品检验研究院,杭州 310052
- 3. 浙江工业大学绿色制药协同创新中心,杭州 310032
- 折叠
摘要
目的 探讨3种光毒性条件下细胞内p38和Erk1/2的表达变化,同时对3种检测方法进行比较,并深入研究其分子机制.方法 使用3种测试方法评估氯丙嗪(chlorpromazine,CPZ)和噻洛芬酸(tiaprofenic acid,TA)的光毒性.随后收集3种测试方法所涉及的生物样本的全细胞裂解物,并利用蛋白质印迹法评估细胞内p38和Erk1/2的表达变化.结果 这3种测试方法均能准确判断CPZ和TA作为光毒性阳性化合物.在3T3 NRU光毒性试验中,与空白组相比,TA和CPZ在光毒性剂量下显著提高了 p38的表达水平(P＜0.05),这一现象在其他测试方法中未观察到.在豚鼠和人工皮肤模型的光毒性试验中,p38和Erk1/2表达模式的变化相似.结论 p38和Erk1/2在3T3NRU光毒性试验中具有明显的剂量依赖性和高度敏感性,可被视为评估3T3 NRU光毒性的潜在分子标志物,但在豚鼠试验和EpiKutis人工皮肤试验中尚未可行.豚鼠和EpiKutis人工皮肤表现出更相似的p38和Erk1/2表达变化,提示EpiKutis人工皮肤模型试验方法可能比3T3 NRU方法能更好地替代动物试验.
Abstract
OBJECTIVE To explore the intracellular expression changes of p38 and Erk1/2 under three phototoxic conditions,compare three detection methods,and further investigate their molecular mechanisms.METHODS The methods involved using three testing approaches to assess the phototoxicity of chlorpromazine(CPZ)and tiaprofenic acid(TA).Subsequently,whole-cell lysates from the biological samples used in the three testing methods were collected,and the changes in the expression levels of intracellular p38 and Erk1/2 were evaluated using Western blotting.RESULTS All three testing methods accurately identified CPZ and TA as phototoxic compounds.In the 3T3 NRU phototoxicity assay,compared to the control group,the expression of p38 significantly increased under phototoxic doses of TA and CPZ(P＜0.05),a phenomenon not observed in other testing methods.In phototoxicity assays using guinea pig and artificial skin models,similar expression pattem changes were observed for p38 and Erk1/2.CONCLUSION p38 and Erk1/2 have obvious dose dependence and high sensitivity in the 3T3 NRU phototoxicity test,and can be considered as potential molecular markers for evaluating the phototoxicity of 3T3 NRU.However,they have not been feasible in guinea pig tests and EpiKutis artificial skin tests.Guinea pigs and EpiKutis artificial skin exhibit more similar changes in p38 and Erk1/2 expression,suggesting that the EpiKutis artificial skin model test method may be a better altemative to animal testing than the 3T3 NRU method.
关键词
光毒性/p38/Erk1/2/人工皮肤模型Key words
phototoxicity/p38/Erk1/2/artificial skin model引用本文复制引用
基金项目
浙江省药品监管系统科技计划项目(2021025)
出版年
2024
NETL
NSTL
万方数据