两种原位DNA断链标记技术检测组织切片和培养细胞中的凋亡细胞

Detection of apoptotic cells in tissue sections and cultural cells by tow in situ DNA fragmentation labeling techniques

目的开发应用范围广，特异性、敏感性高的凋亡细胞检测技术。方法用末端脱氧核苷酸转移酶介导ｄＵＴＰ缺口末端标记法（ＴＵＮＥＬ）和Ｋｌｅｎｏｗ大片段介导的原位缺口翻译法（ＩＳＮＴ）检测了甲基泼尼松龙处理的ＳＤ大鼠胸腺组织，正常鼠肠粘膜组织、肿瘤组织等标本的石蜡组织切片，和体外培养的肿瘤细胞药物处理后发生凋亡的细胞爬片，以及再生障碍性贫血（再障）病人的骨髓涂片。结果在大鼠胸腺组织中可见大量凋亡的Ｔ淋巴细胞，鼠肠粘膜组织粘膜上皮细胞以凋亡的方式更新脱落，肿瘤组织中可见散在的凋亡细胞。药物处理的肿瘤细胞和再障病人骨髓细胞中有较多的凋亡细胞。被染成棕色或棕褐色的细胞在形态学上符合凋亡细胞的形态学改变。结论此两种方法可用于检测组织切片和培养细胞中的凋亡细胞，可以检测早期的细胞凋亡、并可通过计算标记指数对凋亡细胞进行定量或半定量。

Objective To use tow in situ DNA fragmentation labeling techniques to detect apoptotic cells in paraffinembedded tissue sections and cultural cells.Methods Specimens of thymus tissue of SD rat treated with methylprednisone succinate, normal intestinal mucous tissue, tumor tissue in paraffinembedded tissue sections and specimens of tumor cells treated with drugs and myeloid cells of aplastic anemia patients in cell smear were detected by terminal deoxynucleotidyl transferasemediated dUTP nick endlabeling(TUNEL)and in situ nick translation(ISNT). Results Rat thymus tissue treated with methylprednisone succinate showed large amounts of apoptotic T lymphocyets. Epithelium of intestinal tissue dropped in form of apoptosis during metabolism. Scattered apoptotic cells observed in tumor tissue. There were many apoptotic cells in tumor cells treated with drugs and myeloid cells of aplastic anemia patients in cell smear. The cells stained by brown were consonant with changes of apoptotic cells in morphology. Conclusions The methods can detect apoptotic cells, especially early apoptotic cells, in paraffinembedded tissue sections and cultural cells, and have quantity or semiquantity of apoptotic cells by counting apoptotic index. The assay is one of preferred methods for apoptosis in vivo. 