细胞凋亡率检测方法的探索  

Exploration of techniques for comprehensive assessment of cell apoptosis rate

作　　者：郭茜茜[1] 于广[1] 吴丹丹[1] 苗苗[1] 佟伟[1] 王光川[1] 

机构地区：[1]辽宁医学院免疫教研室,锦州121000

出　　处：《免疫学杂志》2015年第5期425-429,共5页Immunological Journal

摘　　要：目的探索可用于全面准确评价细胞凋亡率的检测技术。方法选取地塞米松诱导胸腺细胞凋亡模型,用流式细胞术及DNA琼脂糖凝胶电泳等方法对处于不同凋亡阶段的胸腺细胞凋亡率进行检测分析。结果流式细胞分析对于早期凋亡胸腺细胞的检测不仅具有高度灵敏性,而且可有效区分凋亡与坏死细胞,但对于凋亡晚期细胞膜严重受损乃至破碎的细胞,单纯流式细胞仪检测不能有效反映真实凋亡率;借助琼脂糖凝胶电泳法可检测破碎后释放到胸腺组织或培养基内的DNA碎片;当凋亡严重导致大量细胞破碎降解时,对样本进行细胞计数比较亦有重要参考价值。结论为全面和准确地评估细胞凋亡率,需选择多种检测方法结合分析,细胞学及寡核苷酸片段检测技术是一个相当不错的选择。To explore the techniques that can be used to accurately and comprehensively assess the rate of cell death, we employed a mouse model of dexamethasone-induced thymocyte apoptosis. The rate of various stages of apoptosis was detected and analyzed over the course of 24 hours by using both flow cytometry and DNA agarose gel electrophoresis. Not only was detection of apoptosis by flow eytometry highly sensitive, it was also efficient to distinguish apoptotic from necrotic cells. However, flow cytometry alone was not effective at distinguishing the rate of apoptosis especially for the damaged and fragmented cells. In this scenario, agarose gel electrophoresis could be adopted to detect the DNA fragments released from the dying cells. During cases of large scale apoptosis and extensive cell degradation, cell counts would be of extra value. Taken together, to comprehensively and accurately assess the rate of apoptosis, a combined approach utilizing both flow eytometry and gel electrophoresis are necessary to quantify both cellular and oligonucleotide fragments, respectively.

关 键 词：细胞凋亡率 琼脂糖凝胶电泳法 流式细胞术 

分 类 号：R446.6[医药卫生—诊断学]