Department of Pharmacology, Shantou University Medical College, PR China.
Biochem Pharmacol. 2010 May 15;79(10):1428-36. doi: 10.1016/j.bcp.2010.01.021. Epub 2010 Jan 25.
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.
N-正丁基氟哌啶醇碘化物（F2），一种源自氟哌啶醇的新型化合物，可在体外和体内防止缺血/再灌注（I/R）损伤的破坏性影响。我们测试了 F2 对心肌细胞缺氧/复氧（H/R）损伤的心肌保护作用是否通过调节原代培养心肌细胞中的蛋白激酶 C（PKC）活性来介导。心室心肌细胞的原代培养物经历 2 小时的缺氧和 30 分钟的复氧。测量总 PKC 活性，并通过分段 Western blot 分析评估 PKCalpha、betaII、delta 和 epsilon 同工型的易位模式。我们研究了在存在和不存在特异性抑制剂和激活剂的情况下，PKC 同工型易位与 H/R 诱导损伤之间的关联。测量包括通过肌酸激酶（CK）释放评估细胞损伤，以及通过 Annexin V-FITC 测定法测量凋亡。在暴露于 H/R 的原代培养心肌细胞中，PKCalpha、delta 和 epsilon 易位，PKCbetaII 活性没有变化。总 PKC 活性、CK 释放和凋亡在 H/R 后增加。用传统的 PKC 抑制剂 Go6976 处理可降低早期生长反应-1（Egr-1）蛋白表达并减轻凋亡。PKCepsilon 抑制剂肽 epsilonV1-2 增加 H/R 损伤而不影响 Egr-1 表达。F2 预处理可抑制 PKCalpha 的易位，增加 PKCepsilon 的易位，并缓解 CK 释放和凋亡。F2 的保护作用部分被传统的 PKC 激活剂百里香酚（TXA）和 epsilonV1-2 肽阻断。F2 显著减轻 H/R 诱导的损伤，这可能归因于抑制 PKCalpha 和激活 PKCepsilon 的综合益处。
