Site-specific protein modification by 3-n-butylphthalide in primary hepatocytes: Covalent protein adducts diminished by glutathione and N-acetylcysteine

doi:10.1016/j.lfs.2021.120125

	Site-specific protein modification by 3-n-butylphthalide in primary hepatocytes: Covalent protein adducts diminished by glutathione and N-acetylcysteine
	Xue, Yaru 2; Ren, Xuelian 2; Zhu, Zhengdan 3,4; Lei, Peng 2; Liu, Mengling 2,5; Wan, Mimi 1; Zhong, Dafang 2; Huang, He 2; Diao, Xingxing 2
刊名	LIFE SCIENCES
	2021-12-15
卷号	287 页码:14
关键词	3-n-butylphthalide Hepatotoxicity Oxidative stresses Covalent protein modification Drug-drug interaction
ISSN号	0024-3205
DOI	10.1016/j.lfs.2021.120125
通讯作者	Diao, Xingxing(xxdiao@simm.ac.cn)
英文摘要	Aims: 3-n-Butylphthalide (NBP) is widely used for the treatment of cerebral ischaemic stroke but can causeliver injury in clinical practice. This study aims to elucidate the underlying mechanisms and propose potential preventive strategies. Main methods: NBP and its four major metabolites, 3-hydroxy-NBP (3-OH-NBP), 10-hydroxy-NBP, 10-keto-NBP and NBP-11-oic acid, were synthesized and evaluated in primary human or rat hepatocytes (PHHs, PRHs). NBPrelated substances or amino acid adducts were identified and semi-quantitated by ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS). The target proteins and binding sites were identified by shotgun proteomics based on peptide mass fingerprinting coupled with tandem mass spectrometry and verified by molecular docking. Key findings: The toxicity of NBP and its four major metabolites were compared in both PHHs and PRHs, and 3OH-NBP was found to be the most toxic metabolite. 3-OH-NBP induced remarkable cell death and oxidative stresses in hepatocytes, which correlated well with the levels of glutathione and N-acetylcysteine adducts (3GSH-NBP and 3-NAC-NBP) in cell supernatants. Additionally, 3-OH-NBP covalently conjugated with intracellular Cys, Lys and Ser, with preferable binding to Cys sites at Myh9 Cys1380, Prdx4 Cys53, Vdac2 Cys48 and Vdac3 Cys36. Furthermore, we found that CYP3A4 induction by rifampicin augmented NBP-induced cell toxicity and supplementing with GSH or NAC alleviated the oxidative stresses and reactive metabolites caused by 3-OH-NBP. Significance: Our work suggests that glutathione depletion, mitochondrial injury and covalent protein modification are the main causes of NBP-induced hepatotoxicity, which may be prevented by exogenous GSH or NAC supplementation and avoiding concomitant use of CYP3A4 inducers.
资助项目	National Outstanding Youth Science Fund Project of Natural Science Foundation of China[81903701] ; National Natural Science Foundation of China[81973164] ; Shanghai Pujiang Program[19PJ1411200]
WOS关键词	LIPID-PEROXIDATION ; LIVER ; RAT ; ACETAMINOPHEN ; DEPLETION ; METABOLITES ; MECHANISMS ; TOXICITY ; SULFOTRANSFERASE ; ELECTROPHILES
WOS研究方向	Research & Experimental Medicine ; Pharmacology & Pharmacy
语种	英语
出版者	PERGAMON-ELSEVIER SCIENCE LTD
WOS记录号	WOS:000728125600003
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/299208]
专题	中国科学院上海药物研究所
通讯作者	Diao, Xingxing
作者单位	1.Waters Technol Shanghai Co Ltd, Shanghai 201203, Peoples R China 2.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China 3.Peking Univ, Acad Adv Interdisciplinary Studies, Beijing 100871, Peoples R China 4.Beijing Inst Big Data Res, Beijing 100871, Peoples R China 5.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Xue, Yaru,Ren, Xuelian,Zhu, Zhengdan,et al. Site-specific protein modification by 3-n-butylphthalide in primary hepatocytes: Covalent protein adducts diminished by glutathione and N-acetylcysteine[J]. LIFE SCIENCES,2021,287:14.
APA	Xue, Yaru.,Ren, Xuelian.,Zhu, Zhengdan.,Lei, Peng.,Liu, Mengling.,...&Diao, Xingxing.(2021).Site-specific protein modification by 3-n-butylphthalide in primary hepatocytes: Covalent protein adducts diminished by glutathione and N-acetylcysteine.LIFE SCIENCES,287,14.
MLA	Xue, Yaru,et al."Site-specific protein modification by 3-n-butylphthalide in primary hepatocytes: Covalent protein adducts diminished by glutathione and N-acetylcysteine".LIFE SCIENCES 287(2021):14.