title: Enhanced expression of WD repeat-containing protein 35 via CaMKK/AMPK activation in bupivacaine-treated Neuro2a cells.
abstract: We previously reported that bupivacaine induces reactive oxygen species (ROS) generation, p38 mitogen-activated protein kinase (MAPK) activation and nuclear factor-kappa B activation, resulting in an increase in expression of WD repeat-containing protein 35 (WDR35) in mouse neuroblastoma Neuro2a cells. However, the identity of signaling upstream of p38 MAPK pathways to WDR35 expression remains unclear. It has been shown that AMP-activated protein kinase (AMPK) can activate p38 MAPK through diverse mechanisms. In addition, several kinases acting upstream of AMPK have been identified including Ca2+/calmodulin-dependent protein kinase kinase (CaMKK). Recent studies reported that AMPK may be involved in bupivacaine-induced cytotoxicity in Schwann cells and in human neuroblastoma SH-SY5Y cells. The present study was undertaken to test whether CaMKK and AMPK are involved in bupivacaine-induced WDR35 expression in Neuro2a cells. Our results showed that bupivacaine induced activation of AMPK and p38 MAPK in Neuro2a cells. The AMPK inhibitors, compound C and iodotubercidin, attenuated the bupivacaine-induced activation of AMPK and p38 MAPK, resulting in an inhibition of the bupivacaine-induced increase in WDR35 expression. Treatment with the CaMKK inhibitor STO-609 also attenuated the bupivacaine-induced activation of AMPK and p38 MAPK, resulting in an inhibition of the bupivacaine-induced increase in WDR35 expression. These results suggest that bupivacaine activates AMPK and p38 MAPK via CaMKK in Neuro2a cells, and that the CaMKK/AMPK/p38 MAPK pathway is involved in regulating WDR35 expression.
authors.affiliation: Department of Pharmacology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan; Department of Anesthesiology, Renmin Hospital of Wuhan University, Wuhan, Hubei, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Huang
authors.affiliation: Department of Pharmacology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan.
authors.forename: Fumio
authors.initials: F
authors.lastname: Kondo
authors.affiliation: Advanced Medical Research Center, Aichi Medical University, Nagakute, Aichi, Japan.
authors.forename: Masahiko
authors.initials: M
authors.lastname: Gosho
authors.affiliation: Department of Pharmacology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan.
authors.forename: Guo-Gang
authors.initials: GG
authors.lastname: Feng
authors.affiliation: Department of Anesthesiology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan.
authors.forename: Misako
authors.initials: M
authors.lastname: Harato
authors.affiliation: Department of Anesthesiology, Renmin Hospital of Wuhan University, Wuhan, Hubei, China.
authors.forename: Zhong-yuan
authors.initials: ZY
authors.lastname: Xia
authors.affiliation: Department of Pharmacology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan.
authors.forename: Naohisa
authors.initials: N
authors.lastname: Ishikawa
authors.affiliation: Department of Anesthesiology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Fujiwara
authors.affiliation: Department of Pharmacology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan.
authors.forename: Shoshiro
authors.initials: S
authors.lastname: Okada
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D000779:Anesthetics, Local / Q000494:pharmacology*; D000818:Animals; D002045:Bupivacaine / Q000494:pharmacology*; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D003598:Cytoskeletal Proteins; D004789:Enzyme Activation / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects*; D053823:Hedgehog Proteins; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D020935:MAP Kinase Signaling System / Q000187:drug effects; D051379:Mice; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism
chemical_list: D000779:Anesthetics, Local; D003598:Cytoskeletal Proteins; D053823:Hedgehog Proteins; D047908:Intracellular Signaling Peptides and Proteins; D011506:Proteins; C558596:WDR35 protein, human; C558597:WDR35 protein, mouse; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase; D048051:p38 Mitogen-Activated Protein Kinases; D055372:AMP-Activated Protein Kinases; D002045:Bupivacaine