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采用荧光显微镜、电子显微镜、琼脂糖凝胶电泳、流式细胞仪分析等技术研究鱼呼肠孤病毒诱导草鱼肾细胞 (CIK)凋亡。结果显示 ,鱼呼肠孤病毒感染CIK细胞后 ,光镜下可见空斑形成 ;荧光染色观察到细胞凋亡碎裂核 ;且电镜下呈现细胞核裂解 ,核周裂隙增大 ,细胞膜内陷并出泡形成凋亡小体现象 ;琼脂糖凝胶电泳出现 1 80— 2 0 0bp整数倍的DNA梯形带 ;流式细胞仪检测到典型的细胞凋亡峰 ,在病毒感染 48h ,细胞凋亡百分率达 1 5%。上述结果显示 ,鱼呼肠孤病毒诱导CIK细胞凋亡是其致细胞病变死亡的重要表现形式之一。
Fluorescent microscopy, electron microscopy, agarose gel electrophoresis, flow cytometry and other techniques to study fish reovirus induced apoptosis of grass carp kidney cells (CIK). The results showed that plaque formation was observed under light microscopy after CIK cells were infected by fish reovirus. Apoptotic fissure nuclei were observed by fluorescence staining. The cytoplasmic nuclei were cleaved by electron microscopy, The apoptotic bodies were observed by flow cytometry. DNA ladder with an integer multiple of 1 80-20000 bp was obtained by agarose gel electrophoresis. Apoptotic peak was detected by flow cytometry. The percentage of apoptotic cells reached 48 h 15%. The above results show that fish reovirus induced apoptosis of CIK cells is one of the important manifestations of the death of cytopathic effect.