title: Danshen (Salvia miltiorrhiza) water extract inhibits paracetamol-induced toxicity in primary rat hepatocytes via reducing CYP2E1 activity and oxidative stress.
abstract: OBJECTIVE
  This study aimed to investigate the protective effects of Danshen (Salvia miltiorrhiza) water extract (DSE) and its major phenolic acid components against CYP2E1-mediated paracetamol (APAP)-induced hepatic toxicity.
  
  
  METHODS
  The protection and underlying mechanisms were detected in CYP2E1 overexpression primary rat hepatocytes by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, alamar blue assay, CYP2E1 inhibition assay and glutathione assay.
  
  
  RESULTS
  After APAP treatment, DSE (0.06-1 mg/ml) significantly increased cell viability in MTT assay. Two major components danshensu (8.2-130.5 μm) and salvianolic acid B (Sal B; 3.3-53.5 μm) mainly contributed to this protection, but rosmarinic acid, protocatechuic aldehyde and Sal A did not. Alamar blue assay showed that DSE, danshensu and Sal B maintained mitochondrial metabolic activity. DSE inhibited CYP2E1 (Ki = 1.46 mg/ml) in a mixed mode in rat liver microsomes in vitro; DSE decreased APAP-induced total glutathione depletion and preserved redox status (GSH/GSSG ratio) in hepatocytes. Danshensu and Sal B did not inhibit CYP2E1 or decrease total glutathione depletion, but preserved redox status.
  
  
  CONCLUSIONS
  DSE protected hepatocytes against APAP-induced injury via maintenance of mitochondrial metabolic activity, CYP2E1 inhibition, reduction of total glutathione depletion and preservation of redox status. Danshensu and Sal B were mainly responsible for this protection.
authors: School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China.
authors: Xuelin
authors: http://orcid.org/0000-0002-4051-7851
authors: X
authors: Zhou
authors: School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China.
authors: Ching Mei
authors: CM
authors: Cheung
authors: School of Chinese Medicine, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China.
authors: Jia-Ming
authors: JM
authors: Yang
authors: School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China.
authors: Penelope M Y
authors: PM
authors: Or
authors: Department of Orthopaedics & Traumatology, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China.
authors: Wayne Y W
authors: WY
authors: Lee
authors: School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China.
authors: John H K
authors: JH
authors: Yeung
mesh_terms: D000082:Acetaminophen / Q000494:pharmacology*; D000818:Animals; D001572:Benzofurans / Q000737:chemistry / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D019392:Cytochrome P-450 CYP2E1 / Q000378:metabolism*; D004365:Drugs, Chinese Herbal / Q000737:chemistry / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D062385:Hydroxybenzoates / Q000737:chemistry / Q000494:pharmacology; D007773:Lactates / Q000737:chemistry / Q000494:pharmacology; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D027561:Salvia miltiorrhiza / Q000737:chemistry*; D014867:Water / Q000737:chemistry*; D053630:Depsides
chemical_list: D000082:Acetaminophen; D001572:Benzofurans; D019392:Cytochrome P-450 CYP2E1; D004365:Drugs, Chinese Herbal; D005978:Glutathione; D062385:Hydroxybenzoates; D007773:Lactates; D010936:Plant Extracts; D014867:Water; C028972:dan-shen root extract; C035055:3,4-dihydroxyphenyllactic acid; C076944:salvianolic acid B; D053630:Depsides; C017616:phenolic acid
keywords: CYP2E1; Salvia miltiorrhiza; oxidative stress; paracetamol; primary rat hepatocytes