DNA末端原位标记结合免疫酶标技术研究骨髓增生异常综合征原位细胞凋亡

目的　观察骨髓增生异常综合征 (MDS)患者原位细胞凋亡数量及红系、巨核系凋亡状况。方法　用DNA末端原位标记 (ISEL)结合碱性磷酸酶 抗碱性磷酸酶 (APAAP)免疫酶标技术分析 2 5例MDS患者骨髓塑料冷包埋切片中造血细胞原位凋亡数量及红系和巨核系凋亡状况 ,缺铁性贫血 14例作对照。结果　MDS患者凋亡细胞数平均为 (39.44± 2 9.34 ) /mm2 ,对照组为 (13.43± 8.39) /mm2 (P<0 .0 1) ;MDS中低危组 (RA +RAS)凋亡细胞数为 (4 7.5 6± 32 .86 ) /mm2 ,较高危组 (RAEB +RAEB t)[(2 1.87± 13.6 5 ) /mm2 ]高 (P <0 .0 5 )。MDS患者红系、巨核系细胞凋亡分别占该两系细胞的 3.3%和4 5 % ;对照组分别为 3.0 %和 2 .8% ,两组比较差异无显著性。部分显示红系、巨核系形态学特征的凋亡细胞不表达簇分化抗原。结论　MDS存在过度凋亡 ;MDS中危组细胞凋亡数高于高危组 ;未观察到有核红细胞及巨核细胞凋亡明显增加 ,可能与晚期凋亡细胞表面抗原丢失有关

Study on apoptosis in myelodysplastic syndromes by DNA in situ end labelling combined with APAAP

OBJECTIVE: To investigate the total in situ apoptotic cell number and the apoptotic situation in erythroid cell and megakaryocytes in patients with myelodysplastic syndromes (MDS). METHODS: Apoptosis cell number and the apoptotic situation of erythroid cell and megakaryocytes were analysed on cold embedded bone marrow sections from 25 MDS patients by DNA in situ end labelling (ISEL)/alkaline phosphatase anti-alkaline phosphatase (APAAP) double stained techniques. Fourteen cases of iron deficiency anemia (IDA) were taken as control. RESULTS: Mean apoptotic cell numbers in MDS and control group were (39.44 +/- 29.34)/mm(2) and (13.43 +/- 8.39)/mm(2) respectively (P < 0.01). RA/RAS subtypes had a higher apoptosis ratio (47.56 +/- 32.86/mm(2)) than that in RAEB/RAEB-t subtypes (21.87 +/- 13.65/mm(2)) (P < 0.05). Double staining showed similar apoptosis percentage in erythroid cell and megakaryocytes in MDS patients comparing with that of controls (P > 0.05). Some apoptotic cells showing erythroid or megakaryocytic morphologic characteristics expressed no cluster differentiation antigen. CONCLUSION: Overapoptosis existed in MDS, RA/RAS group had a higher apoptosis ratio than RAEB/RAEB-t group. No obvious increased apoptosis in erythroid cell and megakaryocytes was observed in MDS perhaps due to the loss of surface antigens in later stages of apoptotic cells.