title: Oxidative stress is involved in Dasatinib-induced apoptosis in rat primary hepatocytes.
abstract: Dasatinib, a multitargeted inhibitor of BCR-ABL and SRC kinases, exhibits antitumor activity and extends the survival of patients with chronic myeloid leukemia (CML) and Philadelphia chromosome-positive acute lymphoblastic leukemia (ALL). However, some patients suffer from hepatotoxicity, which occurs through an unknown mechanism. In the present study, we found that Dasatinib could induce hepatotoxicity both in vitro and in vivo. Dasatinib reduced the cell viability of rat primary hepatocytes, induced the release of alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) in vitro, and triggered the ballooning degeneration of hepatocytes in Sprague-Dawley rats in vivo. Apoptotic markers (chromatin condensation, cleaved caspase-3 and cleaved PARP) were detected to indicate that the injury induced by Dasatinib in hepatocytes in vitro was mediated by apoptosis. This result was further validated in vivo using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assays. Here we found that Dasatinib dramatically increased the level of reactive oxygen species (ROS) in hepatocytes, reduced the intracellular glutathione (GSH) content, attenuated the activity of superoxide dismutase (SOD), generated malondialdehyde (MDA), a product of lipid peroxidation, decreased the mitochondrial membrane potential, and activated nuclear factor erythroid 2-related factor 2 (Nrf2) and mitogen-activated protein kinases (MAPK) related to oxidative stress and survival. These results confirm that oxidative stress plays a pivotal role in Dasatinib-mediated hepatotoxicity. N-acetylcysteine (NAC), a typical antioxidant, can scavenge free radicals, attenuate oxidative stress, and protect hepatocytes against Dasatinib-induced injury. Thus, relieving oxidative stress is a viable strategy for reducing Dasatinib-induced hepatotoxicity.
authors.affiliation: Institute of Pharmacology & Toxicology, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Xue
authors.forename: Peihua
authors.initials: P
authors.lastname: Luo
authors.forename: Hong
authors.initials: H
authors.lastname: Zhu
authors.forename: Yuqin
authors.initials: Y
authors.lastname: Zhao
authors.forename: Honghai
authors.initials: H
authors.lastname: Wu
authors.forename: Renhua
authors.initials: R
authors.lastname: Gai
authors.forename: Youping
authors.initials: Y
authors.lastname: Wu
authors.forename: Bo
authors.initials: B
authors.lastname: Yang
authors.forename: Xiaochun
authors.initials: X
authors.lastname: Yang
authors.forename: Qiaojun
authors.initials: Q
authors.lastname: He
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002469:Cell Separation; D004247:DNA / Q000737:chemistry; D000069439:Dasatinib; D016521:Electrophoresis, Gel, Pulsed-Field; D005456:Fluorescent Dyes; D022781:Hepatocytes / Q000187:drug effects*; D020287:In Situ Nick-End Labeling; D007211:Indoles; D007770:L-Lactate Dehydrogenase / Q000097:blood; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008930:Mitochondria, Liver / Q000187:drug effects; D008933:Mitochondrial Swelling / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects*; D047428:Protein Kinase Inhibitors / Q000633:toxicity*; D011743:Pyrimidines / Q000633:toxicity*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D013844:Thiazoles / Q000633:toxicity*
chemical_list: D000975:Antioxidants; D005456:Fluorescent Dyes; D007211:Indoles; D047428:Protein Kinase Inhibitors; D011743:Pyrimidines; D017382:Reactive Oxygen Species; D013844:Thiazoles; C007293:DAPI; D004247:DNA; D007770:L-Lactate Dehydrogenase; D000410:Alanine Transaminase; D000069439:Dasatinib; D000111:Acetylcysteine