title: N-n-butyl haloperidol iodide ameliorates cardiomyocytes hypoxia/reoxygenation injury by extracellular calcium-dependent and -independent mechanisms.
abstract: N-n-butyl haloperidol iodide (F2) has been shown to antagonize myocardial ischemia/reperfusion injury by blocking calcium channels. This study explores the biological functions of ERK pathway in cardiomyocytes hypoxia/reoxygenation injury and clarifies the mechanisms by which F2 ameliorates cardiomyocytes hypoxia/reoxygenation injury through the extracellular-calcium-dependent and -independent ERK1/2-related pathways. In extracellularcalcium-containing hypoxia/reoxygenation cardiomyocytes, PKCα and ERK1/2 were activated, Egr-1 protein level and cTnI leakage increased, and cell viability decreased. The ERK1/2 inhibitors suppressed extracellular-calcium-containing-hypoxia/reoxygenation-induced Egr-1 overexpression and cardiomyocytes injury. PKCα inhibitor downregulated extracellularcalcium-containing-hypoxia/reoxygenation-induced increase in p-ERK1/2 and Egr-1 expression. F2 downregulated hypoxia/reoxygenation-induced elevation of p-PKCα, p-ERK1/2, and Egr-1 expression and inhibited cardiomyocytes damage. The ERK1/2 and PKCα activators antagonized F2's effects. In extracellular-calcium-free-hypoxia/reoxygenation cardiomyocytes, ERK1/2 was activated, LDH and cTnI leakage increased, and cell viability decreased. F2 and ERK1/2 inhibitors antagonized extracellular-calcium-free-hypoxia/reoxygenation-induced ERK1/2 activation and suppressed cardiomyocytes damage. The ERK1/2 activator antagonized F2's above effects. F2 had no effect on cardiomyocyte cAMP content or PKA and Egr-1 expression. Altogether, ERK activation in extracellular-calcium-containing and extracellular-calcium-free hypoxia/reoxygenation leads to cardiomyocytes damage. F2 may ameliorate cardiomyocytes hypoxia/reoxygenation injury by regulating the extracellular-calcium-dependent PKCα/ERK1/2/Egr-1 pathway and through the extracellular-calcium-independent ERK1/2 activation independently of the cAMP/PKA pathway or Egr-1 overexpression.
authors.affiliation: Department of Pharmacology, Shantou University Medical College, 22 Xin Ling Road, Shantou, 515041 Guangdong, China.
authors.forename: Yanmei
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Pharmacology, Shantou University Medical College, 22 Xin Ling Road, Shantou, 515041 Guangdong, China.
authors.forename: Gaoyong
authors.initials: G
authors.lastname: Chen
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Southern California, Los Angeles, CA 90033, USA.
authors.forename: Shuping
authors.initials: S
authors.lastname: Zhong
authors.affiliation: Department of Pharmacy, The First Affiliated Hospital, Shantou University Medical College, Shantou 515041, Guangdong, China.
authors.forename: Fuchun
authors.initials: F
authors.lastname: Zheng
authors.affiliation: Department of Pharmacology, Shantou University Medical College, 22 Xin Ling Road, Shantou, 515041 Guangdong, China.
authors.forename: Fenfei
authors.initials: F
authors.lastname: Gao
authors.affiliation: Department of Pharmacology, Shantou University Medical College, 22 Xin Ling Road, Shantou, 515041 Guangdong, China.
authors.forename: Yicun
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Pharmacology, Shantou University Medical College, 22 Xin Ling Road, Shantou, 515041 Guangdong, China.
authors.forename: Zhanqin
authors.initials: Z
authors.lastname: Huang
authors.affiliation: Department of Pharmacology, Shantou University Medical College, 22 Xin Ling Road, Shantou, 515041 Guangdong, China.
authors.forename: Wenfeng
authors.initials: W
authors.lastname: Cai
authors.affiliation: Analytical Cytology Laboratory, Shantou University Medical College, Shantou 515041, Guangdong, China.
authors.forename: Weiqiu
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Pharmacology, Shantou University Medical College, 22 Xin Ling Road, Shantou, 515041 Guangdong, China.
authors.forename: Xingping
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Pharmacology, Shantou University Medical College, 22 Xin Ling Road, Shantou, 515041 Guangdong, China.
authors.forename: Yanshan
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: Department of Pharmacology, Shantou University Medical College, 22 Xin Ling Road, Shantou, 515041 Guangdong, China.
authors.forename: Han
authors.initials: H
authors.lastname: Xu
authors.affiliation: Department of Pharmacology, Shantou University Medical College, 22 Xin Ling Road, Shantou, 515041 Guangdong, China ; Department of Cardiovascular Diseases, The First Affiliated Hospital, Shantou University Medical College, Shantou 515041, Guangdong, China.
authors.forename: Ganggang
authors.initials: G
authors.lastname: Shi
mesh_terms: D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D015687:Cell Hypoxia / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D000242:Cyclic AMP / Q000378:metabolism; D019610:Cytoprotection / Q000187:drug effects; D051766:Early Growth Response Protein 1 / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005110:Extracellular Space / Q000187:drug effects / Q000378:metabolism*; D006220:Haloperidol / Q000031:analogs & derivatives* / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009206:Myocardium / Q000473:pathology; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000201:enzymology / Q000473:pathology*; D010100:Oxygen / Q000494:pharmacology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D019210:Troponin I / Q000378:metabolism; D014700:Verapamil / Q000494:pharmacology
chemical_list: D051766:Early Growth Response Protein 1; C482574:N-n-butyl haloperidol iodide; D047428:Protein Kinase Inhibitors; D019210:Troponin I; D014700:Verapamil; D000242:Cyclic AMP; D007770:L-Lactate Dehydrogenase; D048049:Extracellular Signal-Regulated MAP Kinases; D006220:Haloperidol; D010100:Oxygen; D002118:Calcium