氯丙嗪和噻洛芬酸在三种光毒性测试中致p38和Erk1/2表达变化研究

The Research of Chlorpromazine and Tiaprofenic Acid-induced p38 and Erk1/2 Expression Change in Three Phototoxicity Test Methods

关键词:  光毒性  p38  Erk1/2  人工皮肤模型

Key words:  phototoxicity, p38 MAPK, Erk1/2, artificial skin model

摘要:

目的 深入探讨三种光毒性条件下细胞内p38和Erk1/2的表达变化，同时对三种检测方法进行比较，并深入研究它们的分子机制。方法 使用三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，我们收集了三种测试方法所涉及的生物样本的全细胞裂解物，并利用蛋白免疫印迹（Western Blot，WB）技术评估细胞内p38和Erk1/2的表达变化。结果 这三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3T3 NRU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。尽管如此，与3T3 NRU相比较，豚鼠和Epikutis人工皮肤表现出更为相似的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方法更好的替代动物试验，对豚鼠试验与Epikutis人工皮肤的其他分子标志物的发掘，仍有广阔的前景。

Abstract:

ABSTRACT: OBJECTIVE The study aims 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 Blot (WB) analysis. RESULTS The results indicate that all three testing methods accurately identified CPZ and TA as phototoxic compounds. In the 3T3 NRU phototoxicity assay, it was observed that, compared to the control group, the expression of p38 MAPK 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 pattern changes were observed for p38 and Erk1/2. CONCLUSION This study concludes that the p38 and Erk1/2 can be considered as potential molecular markers for assessing 3T3 NRU phototoxicity, exhibiting dose-dependency and sensitivity. However, feasibility has not been demonstrated in guinea pig and Epikutis artificial skin experiments. Nevertheless, compared to 3T3 NRU, the more similar alterations in p38 and Erk1/2 expression observed in guinea pig and Epikutis artificial skin models suggest that the Epikutis artificial skin model may be a better alternative animal test than the 3T3 NRU phototoxicity, and promising prospects for the exploration of molecular markers in these models.