title: Bupivacaine induces apoptosis via mitochondria and p38 MAPK dependent pathways.
abstract: Mitochondria and the p38 mitogen-activated protein kinase (MAPK) pathways play important roles in apoptosis. Although the effect of bupivacaine on apoptosis is known, it remains unclear whether bupivacaine induces apoptosis via mitochondrial depolarization and the p38 MAPK activity. In this study, SH-SY5Y cells were pretreated respectively with 50μM 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), 10μM 4-(4-Fluorophenyl)-2-[4-(methylsulfinyl)phenyl]-5-(4-pyridyl)-1H-imidazole (SB203580), and 50μM DIDS plus 10μM SB203580 30min prior to the treatment with either 1mM bupivacaine or an equivalent amount of medium. The cell viability, mitochondrial membrane potential, phospho-p38 MAPK (p-p38 MAPK) and cell apoptosis were investigated with MTT assay, western blots, Hoechst 33258 staining and flow cytometry assay. In addition, the roles of chloridion (Cl(-)) channel and reactive oxygen species were studied to explore the molecular mechanism of bupivacaine-induced mitochondrial injury. Pretreatment with DIDS could attenuate reactive oxygen species production, the phosphorylation of p38MAPK, dissipation of mitochondrial membrane potential and apoptosis of SH-SY5Y cells induced by bupivacaine. Pretreatment with SB203580 could attenuate apoptosis, but could not attenuate reactive oxygen species production, or dissipation of mitochondrial membrane potential induced by bupivacaine. These findings indicate that the mitochondrial anion channel and p38 MAPK pathway are implicated in bupicavaine-induced apoptosis. Bupivacaine-induced reactive oxygen species production results in an alteration in the permeability of the mitochondrial membranes and Cl(-) influx into mitochondria, which seems to be responsible for mitochondrial depolarization and the p38 MAPK activation.
authors.affiliation: Dept. of Anesthesiology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong, China. 454921043@qq.com
authors.forename: Jun
authors.initials: J
authors.lastname: Lu
authors.forename: Shi Yuan
authors.initials: SY
authors.lastname: Xu
authors.forename: Qing Guo
authors.initials: QG
authors.lastname: Zhang
authors.forename: Rui
authors.initials: R
authors.lastname: Xu
authors.forename: Hong Yi
authors.initials: HY
authors.lastname: Lei
mesh_terms: D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D000779:Anesthetics, Local / Q000009:adverse effects / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D006690:Bisbenzimidazole / Q000378:metabolism; D002045:Bupivacaine / Q000009:adverse effects / Q000494:pharmacology*; D002460:Cell Line; D002712:Chlorides / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D007093:Imidazoles / Q000494:pharmacology; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D000779:Anesthetics, Local; D002712:Chlorides; D007093:Imidazoles; D010750:Phosphoproteins; D011725:Pyridines; D017382:Reactive Oxygen Species; D048051:p38 Mitogen-Activated Protein Kinases; D006690:Bisbenzimidazole; C093642:SB 203580; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; D002045:Bupivacaine