TUNEL染色结合细胞形态学特点鉴别急性胰腺炎时腺泡细胞的死亡方式

TUNEL staining combined with cell morphologic characteristics for identifying acinar cell death mode in rats with acute pancreatitis

目的:通过脱氧核糖核苷酸末端转移酶介导的缺口末端标记(TdT-mediated dUTP nick labeling,TUNEL)染色检测大鼠急性胰腺炎(acute pancreatitis,AP)中腺泡细胞的死亡情况,探讨TUNEL染色阳性细胞的形态与细胞死亡方式之间的关系.方法:大鼠随机分为ANP组和假手术组(sham operation,SO),每组6只,利用4%的牛磺胆酸钠胰胆管逆行注射建立大鼠急性坏死性胰腺炎(acute necrotizing pancreatitis,ANP)模型,对两组大鼠胰腺组织进行病理学观察并采用TUNEL技术对胰腺组织切片进行染色,光镜下观察TUNEL染色阳性细胞的形态学表现.结果:对比假手术组,ANP组大鼠胰腺组织出现典型的AP样病理改变,同时可见大量的TUNEL染色阳性腺泡细胞.阳性细胞之间的形态学特点各异,其中既有细胞呈凋亡表现,如核固缩和核破裂伴凋亡小体形成;也有细胞呈坏死表现,如核膨胀、核溶解以及胞质空泡化.两种细胞死亡方式的形态学表现之间存在显著差异.结论:AP时,TUNEL染色并不能很好地区分腺泡细胞的坏死与凋亡,但其联合阳性染色细胞的形态学特征可初步鉴别两种细胞死亡方式.

AIM： To detect acinar cell death in rats with acute pancreatitis （AP） by TdT-mediated dUTP nick labeling （TUNEL） staining and to studythe correlation between the morphology of TUNEL positive cells and their death modes. METHODS： Acute necrotizing pancreatitis （ANP） was induced in rats by injection of 4% sodium taurocholate in the pancreaticobiliary duct, and the pathological changes in pancreatic tissue were observed. Pancreatic tissue sections were stained by TUNEL, and the morphological changes of TUNEL positive cells were evaluated under a light microscope. RESULTS： Compared to the sham operated group, the pancreas of ANP rats exhibited typical pathological changes of AP, and had a large number of TUNEL positive acinar cells. The TUNEL positive cells showed various morphologic characteristics, which included the manifestations of both apoptosis and necrosis. The apoptotic morphology was characterized by nuclear condensation and karyorrhexis followed by the formation of apoptotic bodies, while necrotic cells demonstrated nuclear swelling, karyolysis and cytoplasmic vacuolization. There were significant differences between the morphologic characteristics of the two cell death modes. CONCLUSION： TUNEL staining itself cannot well differentiate the apoptosis and necrosis of acinar cells in AP. However, when combined with the morphologic characteristics of TUNEL positive cells, TUNEL staining can preliminarily identify the two modes of cell death.