缺血-再灌注肌肉损伤中程序性细胞死亡的研究

Programmed cell death in skeletal muscle after limb ischemiareperfusion injury

目的：探讨程序性细胞死亡在缺血再灌注肌肉损伤中的作用及缺血再灌注肌肉损伤的发生机制。方法：运用末端脱氧核糖转移酶介导的生物素化脱氧尿嘧啶缺刻标记技术（ＴＵＮＥＬ）观察止血带致大鼠右后肢缺血再灌注腓肠肌细胞程序性死亡的特征与规律，并以左后肢作自身正常对照。结果：正常肌肉肌细胞完整，无程序性死亡细胞；再灌注４小时，肌细胞深度蓝染，程序性死亡细胞率为０．７０±０．０７；再灌注２４小时，肌细胞蓝染程度减轻，其程序性死亡细胞率为０．４８±０．０８，与再灌注４小时比较有极显著性差异（Ｐ＜０．０１）；再灌注２４小时，重度程序性死亡细胞率为０．１２±０．０８，与再灌注４小时（０．３６±０．０９）比较有极显著性差异（Ｐ＜０．０１）。结论：缺血再灌注能致肌肉细胞发生程序性死亡，以再灌注４小时最重。

Objective:To elucidate the potential role of programmed cell death or apoptosis in skeletal muscle after limb ischemiareperfusion injury.Methods:Terminal deoxynucleotidy transferase mediated dUTPbiotin nickend labeling (TUNEL) method was used to determine programmed cell death in gastrocnemius muscle in rats following right hind limb ischemiareperfusion injury.The other hind limb served as controls.Results:In normal muscle,no apoptotic cells could be found.At 4 hours after reperfusion,the incidence of apoptotic cells that stained deeply blue was 0 70±0 07.However,the apoptotic cells were stained lightly,and the incidence of these cells was 0 48±0 08 at 24 hours after reperfusion,which was significantly lower compared to that at 4 hours ( P <0 01).Furthermore,the incidence of severe apoptotic cells was 0 36±0 09 and 0 12±0 08 at 4 and 24 hours following reperfusion respectively,and the difference was significant ( P <0 01).Conclusions:The result indicate that limb ischemiareperfusion can result in programmed cell death in muscular tissue,and apoptotic cells appear to be more common at 4 hours after reperfusion compared to those at 24 hours later.