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) i...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.展开更多
目的观察丹参酮ⅡA(Tanshinone,TanⅡA)联合三氧化二砷(ATO)对人急性早幼粒细胞白血病细胞株(NB4细胞)凋亡的作用;并通过观察两药联合诱导NB4细胞凋亡时p53、Bcl-2表达的变化,探讨其可能的机制。方法通过Annexin V FITC/PI荧光染色观察...目的观察丹参酮ⅡA(Tanshinone,TanⅡA)联合三氧化二砷(ATO)对人急性早幼粒细胞白血病细胞株(NB4细胞)凋亡的作用;并通过观察两药联合诱导NB4细胞凋亡时p53、Bcl-2表达的变化,探讨其可能的机制。方法通过Annexin V FITC/PI荧光染色观察1.0μg·mL-1TanⅡA分别联合0.25、0.5、1.0μmol·L-1的ATO作用于NB4细胞的形态学变化;流式细胞术Annexin V-FITC/PI法测定各组细胞凋亡率;流式细胞仪检测各组细胞p53、Bcl-2的表达。结果 (1)1.0μg·mL-1TanⅡA分别与0.5、1.0μmol·L-1的ATO联合作用NB4细胞24、48和72h的凋亡率均较相应单独用药组凋亡率明显增高(P<0.05);染色荧光显微镜观察l.0μg·mL-1TanⅡA联合1.0μmol·L-1ATO实验组较1.0μmol·L-1ATO对照组72h时更易见到中晚期凋亡细胞和死亡细胞;(2)1.0μg·mL-1TanⅡA与0.5、1.0μmol·L-1的ATO联合作用NB4细胞48h、72h表达p53蛋白的细胞较1.0μmol·L-1ATO单药组明显增多(P<0.05),作用高峰时间为72h;1.0μg·mL-1TanⅡA分别与0.25、0.5、1.0μmol·L-1的ATO联合作用NB4细胞24h时,表达Bcl-2蛋白的细胞数较1.0μmol·L-1ATO单药组显著下降(P<0.05),且与ATO的浓度呈负相关(r=-0.858,P=0.000)。结论联合1.0μg·mL-1TanⅡA可增强0.5、1.0μmol·L-1ATO诱导NB4细胞凋亡;TanⅡA联合ATO诱导NB4细胞p53表达增强和Bcl-2表达减少可能是其促凋亡机制之一。展开更多
基金The National Natural Science Foundation of China, No. 39970561The Key Projects in the National Science and Technology Pillar Program, 2007Z06-017+2 种基金Program for New Century Excellent Talents from Universities, Chinese Ministry of Education, No. NCET-04-0906/NCET-06-0818Fund of the Discipline Leaders of Sichuan Province, No. SZD0418Culture Fund for Excellent Doctoral Dissertations of Sichuan Agricultural University, 2008scybpy-1
文摘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.
文摘目的观察丹参酮ⅡA(Tanshinone,TanⅡA)联合三氧化二砷(ATO)对人急性早幼粒细胞白血病细胞株(NB4细胞)凋亡的作用;并通过观察两药联合诱导NB4细胞凋亡时p53、Bcl-2表达的变化,探讨其可能的机制。方法通过Annexin V FITC/PI荧光染色观察1.0μg·mL-1TanⅡA分别联合0.25、0.5、1.0μmol·L-1的ATO作用于NB4细胞的形态学变化;流式细胞术Annexin V-FITC/PI法测定各组细胞凋亡率;流式细胞仪检测各组细胞p53、Bcl-2的表达。结果 (1)1.0μg·mL-1TanⅡA分别与0.5、1.0μmol·L-1的ATO联合作用NB4细胞24、48和72h的凋亡率均较相应单独用药组凋亡率明显增高(P<0.05);染色荧光显微镜观察l.0μg·mL-1TanⅡA联合1.0μmol·L-1ATO实验组较1.0μmol·L-1ATO对照组72h时更易见到中晚期凋亡细胞和死亡细胞;(2)1.0μg·mL-1TanⅡA与0.5、1.0μmol·L-1的ATO联合作用NB4细胞48h、72h表达p53蛋白的细胞较1.0μmol·L-1ATO单药组明显增多(P<0.05),作用高峰时间为72h;1.0μg·mL-1TanⅡA分别与0.25、0.5、1.0μmol·L-1的ATO联合作用NB4细胞24h时,表达Bcl-2蛋白的细胞数较1.0μmol·L-1ATO单药组显著下降(P<0.05),且与ATO的浓度呈负相关(r=-0.858,P=0.000)。结论联合1.0μg·mL-1TanⅡA可增强0.5、1.0μmol·L-1ATO诱导NB4细胞凋亡;TanⅡA联合ATO诱导NB4细胞p53表达增强和Bcl-2表达减少可能是其促凋亡机制之一。
