Li Menglin, Hu Ting, Tie Cai, Qu Liang, Zheng Hao, Zhang Jinlan
State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, P. R. China.
Pharmaron Beijing Co., Ltd., BDA, Beijing, P.R. China.
Proteomics. 2017 Nov;17(22). doi: 10.1002/pmic.201700001. Epub 2017 Oct 25.
Triptolide (TP), the major active component in Tripterygium wilfordii Hook. f., is widely used for the treatment of rheumatoid arthritis and autoimmune diseases. However, organ toxicity, especially hepatotoxicity and nephrotoxicity, limits its clinical application. To fully understand the mechanism underlying TP toxicity, iTRAQ-based 2D-LC-MS/MS proteomics is used to detect differentially expressed proteins in the livers and kidneys of mice administered the LD50 dose of TP. Functional annotation revealed that multiple pathways are involved in TP toxicity, including acute-phase response signaling, the antigen presentation pathway, FXR/RXR activation, LPS/IL-1-mediated inhibition of RXR function, and EIF2 signaling. Members of the cytochrome P450 protein family that are involved in fatty acid (FA) metabolism, such as CYP2E1, show significant differences in expression among groups. Additionally, the proteomics data suggested that FAs are involved in TP-induced toxicity. FA analysis is conducted using HPLC-MRM to characterize the differences among various groups exposed to TP for different times. It has been found that 20 FAs in the liver show significant differences in abundance among groups, whereas in the kidneys, six FAs show significant differences in abundance. By integrating the proteomic and targeted FA analyses, it has been found that differently expressed proteins and FAs both participate in pathways including cellular lipolytic activity, peroxisomal fatty acid beta-oxidation, and so on. Our data contribute to understanding the mechanisms underlying TP-induced organ toxicity. The results may help to improve the clinical efficacy and safety of TP in the future.
雷公藤甲素（TP）是雷公藤的主要活性成分，广泛用于治疗类风湿性关节炎和自身免疫性疾病。然而，器官毒性，尤其是肝毒性和肾毒性，限制了其临床应用。为了全面了解TP毒性的潜在机制，基于iTRAQ的二维液相色谱-串联质谱蛋白质组学被用于检测给予LD50剂量TP的小鼠肝脏和肾脏中差异表达的蛋白质。功能注释显示，TP毒性涉及多种途径，包括急性期反应信号传导、抗原呈递途径、FXR/RXR激活、LPS/IL-1介导的RXR功能抑制和EIF2信号传导。参与脂肪酸（FA）代谢的细胞色素P450蛋白家族成员，如CYP2E1，在各组之间的表达存在显著差异。此外，蛋白质组学数据表明FA参与了TP诱导的毒性。使用HPLC-MRM进行FA分析，以表征不同时间暴露于TP的各组之间的差异。已发现肝脏中的20种FA在各组之间的丰度存在显著差异，而在肾脏中，6种FA在丰度上存在显著差异。通过整合蛋白质组学和靶向FA分析，发现差异表达的蛋白质和FA都参与了包括细胞脂解活性、过氧化物酶体脂肪酸β-氧化等途径。我们的数据有助于理解TP诱导器官毒性的潜在机制。这些结果可能有助于未来提高TP的临床疗效和安全性。
