芬太尼对乳鼠心肌细胞缺氧复氧损伤的保护作用

The comparison of fentanyl and anoxic preconditioning on the anoxic reoxygenation injury of the neonatal rat cardiomyocytes

目的采用乳鼠心肌细胞缺氧复氧模型,观察芬太尼与缺氧预适应对心肌细胞缺氧复氧损伤的影响,探讨芬太尼能否直接作用于心肌细胞并模拟缺氧预透应(APC)样心肌保护作用,试图在细胞水平上为临床应用大剂量芬太尼减轻心肌缺血再灌注损伤提供理论依据。方法以常规方法制备与培养心肌细胞,分为4组:正常对照组(C组)不经任何处理;单纯缺氧复氧组(A/R组)、缺氧预适应组(AP组)及芬太尼组(F组)均经历2小时缺氧及1小时复氧。缺氧前,AP组预先经历20分钟缺氧及20分钟复氧;F组给予终浓度为50 ng/ml的芬太尼。分别以四甲基偶氮唑盐(MTT)快速比色法检测细胞存活情况、流式细胞仪检测细胞凋亡率及透射电子显微镜观察细胞超微结构改变。结果F组和AP组OD值均显著高于A/R组,细胞凋亡率显著低于A/R组;F组OD值与细胞凋亡率与AP组比较差异无显著性意义;电镜显示A/R组细胞超微结构改变呈典型的凋亡细胞形态学改变,AP组及F组细胞形态大致正常,凋亡细胞少见。结论芬太尼对乳鼠心肌细胞缺氧复氧损伤有保护作用,且其保护效应与APC的心肌保护作用相似。

Objective In the present study, a model of cultured neonatal rat cardiomyocytes was established to observe the influence of fentanyl on the anoxic rcoxygenation injury, and to investigate whether the direct action of fentany at the level of the neonatal rat cardiomyocytes could induce the preconditioning-like effect. Therefore, it would provide evidence, at the level of the myocytes, for the utilization of a large dose of fentanyl in the clinical anesthesia for cardioprotectiun against the ischemic reperfusion injury. Methods Cardiomyocytes were cuhurcd from the neonatal rat heart according to a routine procedure, and divided into four groups： The normal control group（ C group） didn＇t receive any treatment, while the other groups including anoxia/ reoxygenation group（ A/R group）, AP group and fentanyl group（ F group） all received a 2-hour anoxia and 1-hour reoxygenation. Before the prolonged anoxia,AP group was subjected to a 20-min exposure to anoxia followed by a 20-min of reoxygenation, and F group was subjected to a 20-minute exposure to fentanyl （50 ng/ml） followed by a 10-minute drug-free period. The cellular livability（the value of OD） , the apoptosis percentage and the intracellular uhrastructure of the cardiomyocytes were measured by MTT colorimetry, the flow eytometcr and the transmission electron microscope respectively. Results The cellular livability of F group and AP group were significantly higher than A/R group, and the apoptosis percentage of F group and AP group were signifi- cantly lower than A/R group. The cellular uhrastructure of A/R group indicated the feature of apoptosis, and those of AP group and F group remained normally. The livability and the apoptosis percentage of the cardiomyocytes between F group and AP group had no statistical difference. Conclusions Fentanyl can protect the cardiomyocytes against anoxic rcoxygenation injury, and its cardioprotective effect is similar to the effect of IPC.