N-n-Butyl haloperidol iodide protects against hypoxia/reoxygenation-induced cardiomyocyte injury by modulating protein kinase C activity.

Abstract

N-n-Butyl haloperidol iodide (F2), a novel compound derived from haloperidol, protects against the damaging effects of ischemia/reperfusion (I/R) injury in vitro and in vivo. We tested whether the myocardial protection of F2 on cardiomyocyte hypoxia/reoxygenation (H/R) injury is mediated by modulating protein kinase C (PKC) activity in primary cultured cardiomyocytes. Primary cultures of ventricular cardiomyocytes underwent 2-h hypoxia and 30-min reoxygenation. Total PKC activity was measured, and the translocation pattern of PKCalpha, betaII, delta and epsilon isoforms was assessed by fractionated western blot analysis. We investigated the association of PKC isoform translocation and H/R-induced injury in the presence and absence of the specific inhibitors and activator. Measurements included cell damage evaluated by creatine kinase (CK) release, and apoptosis measured by annexin V-FITC assay. In primary cultured cardiomyocytes exposed to H/R, PKCalpha, delta and epsilon were translocated, with no change in PKCbetaII activity. Total PKC activity, CK release and apoptosis were increased after H/R. Treatment with the conventional PKC inhibitor Go6976 reduced early growth response-1 (Egr-1) protein expression and attenuated apoptosis. The PKCepsilon inhibitor peptide epsilonV1-2 increased H/R injury without influencing Egr-1 expression. Pretreatment with F2 inhibited translocation of PKCalpha, increased translocation of PKCepsilon, and relieved the CK release and apoptosis. The protection of F2 was blocked in part by the conventional PKC activator thymeleatoxin (TXA) and epsilonV1-2 peptide. F2 significantly alleviated H/R-induced injury, which might be attributed to the combined benefits of inhibiting PKCalpha and activating PKCepsilon.

• Authors: Jin-Zhi Wang, Cong-Yi Cai, Yan-Mei Zhang, Jin-Hong Zheng, Yi-Cun Chen, Wei-Qiu Li, Gang-Gang Shi
• Journal: Biochemical pharmacology (Biochem Pharmacol) Vol. 79 Issue 10 Pg. 1428-36 (May 15 2010) ISSN: 1873-2968 [Electronic] England
• PMID: 20105432 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: 2010 Elsevier Inc. All rights reserved.

Chemical References

• Cardiotonic Agents
• Early Growth Response Protein 1
• Egr1 protein, rat
• N-n-butyl haloperidol iodide
• Protein Isoforms
• Protein Kinase C
• Haloperidol

Topics

• Animals
• Apoptosis (drug effects)
• Blotting, Western
• Cardiotonic Agents (pharmacology)
• Cells, Cultured
• Early Growth Response Protein 1 (biosynthesis)
• Gene Expression (drug effects)
• Haloperidol (analogs & derivatives, pharmacology)
• Hypoxia (drug therapy, physiopathology)
• Myocytes, Cardiac (drug effects, enzymology, physiology)
• Protein Isoforms
• Protein Kinase C (biosynthesis, drug effects, metabolism)
• Rats
• Rats, Sprague-Dawley
• Up-Regulation (physiology)