口蹄疫病毒诱导宿主细胞凋亡的研究

Studies on Apoptosis of Host Cells Induced by Foot-and-Mouth Disease Virus

本文报道了口蹄疫病毒 (Foot and MouthDiseaseVirus,FMDV)在体外诱导PK 15细胞凋亡的研究结果。采用Hoechst332 5 8荧光探针、DNA凝胶电泳、脱氧核糖核酸转移酶介导的缺口末端标记 (TUNEL)技术均检测到了典型的细胞凋亡。结果显示 :使用感染性滴度为 4.8lgTCID50 /mL的口蹄疫病毒感染PK 15细胞 ,在培养 32h后荧光探针检测呈现典型的凋亡细胞核固缩和梅花状碎裂核 ,并伴随有凋亡小体出现 ,凋亡率约为 2 0 % ;DNA凝胶电泳显示ladder梯带 ;末端标记检测到强绿色荧光标记物结合于凋亡细胞核上。研究结果提示 :口蹄疫病毒可以在体外诱导宿主细胞凋亡 ,细胞凋亡是其致细胞病变死亡的重要途径之一。

Apoptosis of PK 15 cells induced by Foot and Mouth Disease Virus (FMDV) in vitro was reported in this paper. Typical cell apoptosis was detected by use of Hoechst 33258 fluorescence probe, agarose gel electrophoresis and in situ end labeling (TUNEL). After PK 15 cells were infected by titration of 4.8 lg TCID 50 /mL FMDV for 32 h, apoptosis characteristics of nuclear condensation, fragmentation, accompanied by apoptotic bodies formation (Hoechst 33258 staining), 180 200 integer fold sized pieces DNA Ladders (agarose gel electrophoresis) and strong green fluorescence dots (TUNEL) were all exhibited, and cell apoptosis was approximately 20%. In addition, the quantitative analysis of apoptosis in PK 15 cells induced by FMDV showed that apoptosis was correlated with infection of virus, and it was also time dependent. Results indicate that FMDV can induce apoptosis of host cells and apoptosis plays an important role in the cytopathogencity effect of FMDV.