Zhang Sheng, Li Chao, Feng Tingting, Cao Shuai, Zhou Heng, Li Liming, Hu Qing, Mao Xiuhong, Ji Shen
Tianjin University of Traditional Chinese Medicine, Tianjin, 300139, China; NMPA Key Laboratory for Quality Control of Traditional Chinese Medicine, Shanghai Institute for Food and Drug Control, Shanghai, 201203, China.
NMPA Key Laboratory for Quality Control of Traditional Chinese Medicine, Shanghai Institute for Food and Drug Control, Shanghai, 201203, China; Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China.
J Ethnopharmacol. 2021 Jul 15;275:114118. doi: 10.1016/j.jep.2021.114118. Epub 2021 Apr 18.
Realgar, a famous traditional Chinese mineral medicine, has been toxic to the renal system. However, the underlying mechanism of Realgar nephrotoxicity is still unclear.
This study aimed to investigate the potential mechanism of Realgar-induced nephrotoxicity by using a label-free quantitative proteomic method.
36 mice were randomly divided into four groups: Control group, 0.5-, 1.0, and 2.0 g/kg Realgar group. After one week, serum biochemical parameters and renal histopathological examination were performed. Label-free quantitative proteomics was used to identify differentially expressed proteins which were subsequently analyzed with bioinformatics methods. Western blot was utilized to verify the six representative protein expressions.
The results showed that 2.0 g/kg Realgar significantly increased blood urea nitrogen and induced the formation of tube cast of renal tubules, while the lower-dose of 0.5 g/kg and 1.0 g/kg Realgar showed no changes. Label-free proteomic analysis identified 3138 proteins, and 272 of those proteins were screened for significant changes in a dose-dependent manner. Functional enrichment analysis suggested that these proteins could affect the apoptotic process and oxidative stress. Representative proteins in the 2.0 g/kg Realgar group, including Cat, Bad, Cycs, Nqo1, Podxl, and Hmox1, were verified by western blot.
The results in this study suggest that apoptosis and oxidative stress might be related to the Realgar-induced nephrotoxicity in mice. Moreover, the strategy of proteomics could contribute to the understanding of the mechanisms of nephrotoxicity in mice exposed to Realgar.
雄黄是一种著名的传统中药矿物药，对肾脏系统有毒性。然而，雄黄肾毒性的潜在机制仍不清楚。
本研究旨在通过无标记定量蛋白质组学方法探讨雄黄诱导肾毒性的潜在机制。
将36只小鼠随机分为四组：对照组、0.5 g/kg、1.0 g/kg和2.0 g/kg雄黄组。一周后，进行血清生化参数检测和肾脏组织病理学检查。采用无标记定量蛋白质组学技术鉴定差异表达蛋白，随后用生物信息学方法进行分析。利用蛋白质免疫印迹法验证六种代表性蛋白的表达。
结果显示，2.0 g/kg雄黄显著增加血尿素氮水平并诱导肾小管管型形成，而较低剂量的0.5 g/kg和1.0 g/kg雄黄则无此变化。无标记蛋白质组学分析鉴定出3138种蛋白质，其中272种蛋白质呈剂量依赖性显著变化。功能富集分析表明，这些蛋白质可能影响细胞凋亡过程和氧化应激。通过蛋白质免疫印迹法验证了2.0 g/kg雄黄组中的代表性蛋白质，包括过氧化氢酶（Cat）、促凋亡蛋白Bad、细胞色素c（Cycs）、醌氧化还原酶1（Nqo1）、血小板/内皮细胞黏附分子1（Podxl）和血红素加氧酶1（Hmox1）。
本研究结果表明，细胞凋亡和氧化应激可能与雄黄诱导的小鼠肾毒性有关。此外，蛋白质组学策略有助于理解暴露于雄黄的小鼠肾毒性机制。
