中文摘要：

目的：观察新型钙拮抗剂碘化N-正丁基氟哌啶醇（N-n-butyl haloperidol iodide,F2）及经典钙拮抗剂维拉帕米对缺氧/复氧（hypoxia/reoxygenation,H/R）时心肌细胞PKCα/ERK1/2信号通路及损伤的作用。方法：建立新生大鼠心肌细胞H/R模型,于缺氧液和复氧液中加入F2（1×10-6mol/L）及维拉帕米（2×10-6mol/L）。采用Western Blot方法检测心肌细胞磷酸化PKCα（pPKCα）、总PKCα、磷酸化ERK（p-ERK1/2）和总ERK1/2蛋白表达的变化;双抗体夹心光化学测定法检测培养细胞上清液中肌钙蛋白（cardiac troponin I,c Tn I）的含量,Hoechst 33342染色法检测心肌细胞早中期凋亡情况,Cell Counting Kit-8比色法检测心肌细胞存活率。结果：H/R激活培养心肌细胞PKCα和ERK1/2,使细胞c Tn I浓度升高,早中期凋亡增加,存活率下降;PKCα抑制剂可下调H/R所致的PKCα和ERK1/2活化程度;ERK1/2抑制剂可抑制H/R所致心肌细胞c Tn I漏出,减少细胞凋亡,提高细胞存活率;F2及维拉帕米能够下调H/R所致PKCα和ERK1/2的活化程度,抑制细胞c Tn I漏出,减少细胞凋亡,提高细胞存活率;PKCα激动剂和ERK激动剂可拮抗F2对H/R心肌细胞PKCα和ERK1/2激活的抑制作用及对心肌细胞损伤的保护作用。结论：H/R可引起心肌细胞PKCα/ERK1/2通路激活,F2和维拉帕米均可通过调节PKCα/ERK1/2信号通路拮抗心肌细胞H/R损伤。

英文摘要：

AIM： To investigate effects of N-nbutyl haloperidol iodide（ F2） and verapamil,a neotype and a typical calcium channel blocker respecrively,on PKCα / ERK1 /2 signaling pathway and hypoxia / reoxygenation（ H / R） injury of cultured cardiomyocytes. METHODS： The H / R models of neonatal rat cardiomyocytes were established. F2（ 1× 10- 6mol / L） and verapamil（ 2 × 10- 6mol / L）were added to hypoxia buffer and reoxygenation buffer. p-PKCα,total PKCα,p-ERK1 /2 and total ERK protein expressions in cardiomyocytes were examined by western-blot analyses. Dual-antibody sandwich photochemical assay was used to detect cardiac troponin I（ c Tn I） content in cell culture supernatant.Hoechst 33342 staining was used to detect early metaphase apoptosis in cardiomyocytes. Cell Counting Kit-8 colorimetric assay was used to detect cardiomyocyte viability. RESULTS： In H / R cardiomyocytes,PKCα and ERK1 /2 were activated, c Tn I leakage elevated,apoptosis increased,and cell viability decreased. The PKCα inhibitor down-regulated H / R-induced PKCα and ERK1 /2 activation.The ERK1 /2 inhibitors suppressed cellular c Tn I leakage,reduced apoptosis,and improved cell viability caused by H / R. F2 and verapamil down-regulated H / R-induced PKCα and ERK1 /2 activation,inhibited cellular c Tn I leakage,reduced early metaphase apoptosis,and promoted cell survival. The PKCα agonist and ERK agonist were found to antagonize F2＇ inhibitory effects on PKCα and ERK1 /2activation and protective effects in H / R cardiomyocytes. CONCLUSION： H / R leads to cardiomyocytes PKCα / ERK1 /2 signaling pathway activation.F2 and verapamil exerts cardioprotective effects against H / R injury by regulating PKCα / ERK1 /2 signaling pathway.