摘要
目的通过AnnexinⅤ/PI流式细胞分析法和TUNEL法检测肝细胞凋亡,比较两种方法的优缺点。方法以体外培养人肝细胞株HL-7702为研究对象,分别设立TGF-β1组(加入TGFβ-1至终浓度为20 ng/ml,作用72 h后收集细胞进行检测)和正常对照组(加入等量PBS),采用AnnexinⅤ/PI流式细胞分析法和TUNEL法检测肝细胞凋亡。结果AnnexinⅤ/PI流式细胞分析显示,TGFβ-1组与正常对照组相比肝细胞的早期凋亡率和总凋亡率均显著增加[(10.55±4.71)%vs(5.98±2.97)%,P<0.01;(19.94±3.68)%vs(13.27±4.62)%,P<0.05]。TUNEL法检测显示,细胞核中有棕黄色着染者为阳性细胞,细胞呈典型的凋亡形态学改变,即表现为变小、变圆、核固缩;而正常细胞的胞核不着染。经Iimage-Pro Plus图像分析软件处理,显示TGF-β1组与正常对照组相比肝细胞凋亡率显著增加[(30.25±6.43)%vs(4.75±0.96)%,P<0.01]。结论AnnexinⅤ/PI流式细胞分析法特异性高,TUNEL法灵敏度高,两种方法相结合检测细胞凋亡更准确。
Objective To compare the advantages and disadvantages of AnnexinⅤ/PI with flow cytometry and TUNEL in detecting hepatocyte apoptosis. Methods Cultured human hepatocyte line HL-7702 was divided into TGF-β1 group（treated with TGF-β1 20 ng/ml for 72 h）and normal control group（incubated with equal PBS）.Hepatocyte apoptosis was detected by AnnexinⅤ/PI with flow cytometry and TUNEL. Results Compared with normal control group,the percentages of early and total apoptosis significantly increased in TGF-β1 group by AnnexinⅤ/PI[（10.55±4.71）%vs（5.98±2.97）%,P〈0.01;（19.94±3.68）%vs（13.27±4.62）%,P〈0.05].By TUNEL assay,nuclei stained buffy were detected as positive cells.Meanwhile the cells changed into smaller,rounder and nuclear condensation.But normal nuclei were not stained.The percentage of hepatocyte apoptosis in TGF-β1 group was much higher than in normal control group[（30.25±6.43）%vs（4.75±0.96）%,P〈0.01].Conclusion AnnexinⅤ/PI with flow cytometry is more specific,while TUNEL assay is more sensitive.It is more precise to combine AnnexinⅤ/PI with TUNEL.
出处
《山西医科大学学报》
CAS
2008年第5期476-479,共4页
Journal of Shanxi Medical University
基金
国家人事部留学回国人员择优基金资助项目(200499)
山西省自然科学基金资助项目(20051092)
太原市科技明星专项基金资助项目(070204004)
