芬太尼对缺氧复氧损伤相关的心肌细胞凋亡的影响

The influence of fentanyl on the anoxic reoxygenation injury of the neonatal rat cardiomyocytes

目的观察不同剂量芬太尼对心肌细胞缺氧复氧损伤的影响,在细胞水平上为临床应用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。实验分为正常对照组(C组)、单纯缺氧复氧组(A/R组)、各试验组(F1、F2、F3、F4组)。C组不经任何处理。A/R组、各试验组均经历2h缺氧及1h复氧。缺氧前,各试验组分别给予终浓度为2、10、50、250ng/ml的芬太尼。分别以MTT快速比色法检测细胞存活情况(OD值)、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。结果各试验组OD值均显著高于A/R组,细胞凋亡率显著低于A/R组,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,F3组细胞形态大致正常,凋亡细胞少见。F3组(50ng/ml)在所有试验组中OD值最高,细胞凋亡率最低。芬太尼剂量在50ng/ml内,OD值与剂量呈正相关(r=0.720,P<0.01),细胞凋亡率与剂量呈负相关(r=-0.990,P<0.01)。结论①芬太尼对乳鼠心肌细胞缺氧复氧损伤有保护作用;②在一定剂量范围内芬太尼的保护效应与剂量成正相关。

Objective To observe the influence of various doses of fentanyl on the anoxic reoxygenation injury,and provide evidence for the utilization of fentanyl in the clinical anesthesia for cardioprotection against the ischemic reperfusion injury.Methods Cardiomyocytes were cultured from the neonatal rat heart according to a routine procedure.The normal control group（C group）didn＇t receive any treatment,while the other groups including the anoxia/reoxygenation group（A/R group） and the treatment group（F1,F2,F3,F4 group）all received a 2-hour anoxia and 1-hour reoxygenation.Before the prolonged anoxia,the treatment groups were subjected to a 20-minute exposure to fentanyl（2,10,50,250ng/ml respectively）followed by a 10-minute drug-free period.Then the livability（the value of OD）,the apoptosis percentage and the intracellular ultrastructure of the cardiomyocytes were measured with MTT colorimetry,the flow cytometer and the transmission electron microscope respectively.Results The cellular livability of the treatment groups were significantly higher than A/R group,and the apoptosis percentage of the treatment group were significantly lower than A/R group.The cellular ultrastructure of A/R group indicated the feature of apoptosis,and that of F3 group remained normally.Among all the treatment groups,the cellular livability of F3 group was the highest,and the apoptosis percentage of F3 group was the lowest.Within the dose of 50ng/ml,the livability correlated positively with the dose（r=0.720,P〈0.01）,while the apoptosis rate correlated negatively with the dose（r=-0.990,P〈0.01）.Conclusion ①Fentanyl could protect the cardiomyocytes against anoxic reoxygenation injury;②Within a certain dose,the cardioprotective effect of fentanyl correlated positively with the dose.