通过荧光素膜联蛋白V-FITC/PI双标记体外检测新型鹅病毒性肠炎病毒诱导的细胞凋亡
Detection of apoptosis induced by new type gosling viral enteritis virus in vitro through fluorescein annexin V-FITC/PI double labeling.
作者信息
Chen Shun, Cheng An-Chun, Wang Ming-Shu, Peng Xi
机构信息
Avian Diseases Research Centre, College of Veterinary Medicine of Sichuan Agricultural University, Yaan 625014, Sichuan Province, China.
出版信息
World J Gastroenterol. 2008 Apr 14;14(14):2174-8. doi: 10.3748/wjg.14.2174.
AIM
To achieve a better understanding of the pathogenesis of new type gosling viral enteritis virus (NGVEV) and the relationship between NGVEV and host cells.
METHODS
The apoptosis of duck embryo fibroblasts (DEF) induced by NGVEV was investigated by fluorescence-activated cell sorter (FACS) and fluorescence microscope after the cells were stained with Annexin V-FITC and propidium iodide (PI).
RESULTS
By staining cells with a combination of fluorescein annexin V-FITC and PI, it is possible to distinguish and quantitatively analyze non-apoptotic cells (Annexin V-FITC negative/PI negative), early apoptotic cells (Annexin V-FITC positive/PI negative), late apoptotic/necrotic cells (Annexin V-FITC positive/PI positive) and dead cells (Annexin V-FITC negative/PI positive) through flow cytometry and fluorescence microscope. The percentage of apoptotic cells increased with the incubation time and reached a maximum at 120 h after infection, while the percentage of non-apoptotic cells decreased.
CONCLUSION
NGVEV can induce the infected DEF cells to undergo apoptosis and the apoptosis occurs prior to necrosis.
目的
更好地了解新型雏鹅病毒性肠炎病毒（NGVEV）的发病机制以及NGVEV与宿主细胞之间的关系。
方法
在用膜联蛋白V-异硫氰酸荧光素（Annexin V-FITC）和碘化丙啶（PI）对鸭胚成纤维细胞（DEF）进行染色后，通过荧光激活细胞分选仪（FACS）和荧光显微镜研究NGVEV诱导的DEF细胞凋亡情况。
结果
通过用荧光素膜联蛋白V-FITC和PI联合对细胞进行染色，能够通过流式细胞术和荧光显微镜区分并定量分析非凋亡细胞（Annexin V-FITC阴性/PI阴性）、早期凋亡细胞（Annexin V-FITC阳性/PI阴性）、晚期凋亡/坏死细胞（Annexin V-FITC阳性/PI阳性）和死亡细胞（Annexin V-FITC阴性/PI阳性）。凋亡细胞的百分比随孵育时间增加，在感染后120小时达到最高，而非凋亡细胞的百分比则下降。
结论
NGVEV可诱导被感染的DEF细胞发生凋亡，且凋亡先于坏死发生。
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