Xie Ming-Zhang, Liu Jun-Li, Gao Qing-Zu, Bo De-Ying, Wang Lei, Zhou Xiao-Chun, Zhao Meng-Meng, Zhang Yu-Chao, Zhang Yu-Jing, Zhao Guo-An, Jiao Lu-Yang
Department of Genetics, First Affiliated Hospital of Xinxiang Medical University, Xinxiang, 453000, Henan, China.
Henan Key Laboratory of Neurorestoratology, Henan International Joint Laboratory of Neurorestoratology for Senile Dementia, The First Affiliated Hospital of Xinxiang Medical University, Weihui, 453100, Henan, People's Republic of China.
Clin Proteomics. 2022 Aug 24;19(1):33. doi: 10.1186/s12014-022-09369-7.
Crotonaldehyde (CRA)-one of the major environmental pollutants from tobacco smoke and industrial pollution-is associated with vascular injury (VI). We used proteomics to systematically characterize the presently unclear molecular mechanism of VI and to identify new related targets or signaling pathways after exposure to CRA. Cell survival assays were used to assess DNA damage, whereas oxidative stress was determined using colorimetric assays and by quantitative fluorescence study; additionally, cyclooxygenase-2, mitogen-activated protein kinase pathways, Wnt3a, β-catenin, phospho-ErbB2, and phospho-ErbB4 were assessed using ELISA. Proteins were quantitated via tandem mass tag-based liquid chromatography-mass spectrometry and bioinformatics analyses, and 34 differentially expressed proteins were confirmed using parallel reaction monitoring, which were defined as new indicators related to the mechanism underlying DNA damage; glutathione perturbation; mitogen-activated protein kinase; and the Wnt and ErbB signaling pathways in VI based on Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and protein-protein interaction network analyses. Parallel reaction monitoring confirmed significant (p < 0.05) upregulation (> 1.5-fold change) of 23 proteins and downregulation (< 0.667-fold change) of 11. The mechanisms of DNA interstrand crosslinks; glutathione perturbation; mitogen-activated protein kinase; cyclooxygenase-2; and the Wnt and ErbB signaling pathways may contribute to VI through their roles in DNA damage, oxidative stress, inflammation, vascular dysfunction, endothelial dysfunction, vascular remodeling, coagulation cascade, and the newly determined signaling pathways. Moreover, the Wnt and ErbB signaling pathways were identified as new disease pathways involved in VI. Taken together, the elucidated underlying mechanisms may help broaden existing understanding of the molecular mechanisms of VI induced by CRA.
巴豆醛（CRA）是烟草烟雾和工业污染产生的主要环境污染物之一，与血管损伤（VI）有关。我们利用蛋白质组学系统地表征目前尚不清楚的VI分子机制，并识别暴露于CRA后新的相关靶点或信号通路。细胞存活试验用于评估DNA损伤，而氧化应激则通过比色法和定量荧光研究来测定；此外，使用酶联免疫吸附测定法评估环氧合酶-2、丝裂原活化蛋白激酶途径、Wnt3a、β-连环蛋白、磷酸化表皮生长因子受体2和磷酸化表皮生长因子受体4。通过基于串联质量标签的液相色谱-质谱联用和生物信息学分析对蛋白质进行定量，并使用平行反应监测确认了34种差异表达蛋白，这些蛋白被定义为与VI中DNA损伤、谷胱甘肽紊乱、丝裂原活化蛋白激酶以及Wnt和表皮生长因子受体信号通路潜在机制相关的新指标。平行反应监测确认23种蛋白显著上调（p < 0.05）（变化> 1.5倍），11种蛋白下调（变化< 0.667倍）。DNA链间交联、谷胱甘肽紊乱、丝裂原活化蛋白激酶、环氧合酶-2以及Wnt和表皮生长因子受体信号通路的机制可能通过它们在DNA损伤、氧化应激、炎症、血管功能障碍、内皮功能障碍、血管重塑、凝血级联反应以及新确定的信号通路中的作用导致VI。此外，Wnt和表皮生长因子受体信号通路被确定为参与VI的新疾病通路。综上所述，阐明的潜在机制可能有助于拓宽对CRA诱导VI分子机制的现有认识。
