目的:探讨Annexin V FITC/PI流式细胞术在检测姜黄素诱导的肝星状细胞(HSC)凋亡中的应用。方法:不同浓度姜黄素作用肝星状细胞株HSC-T6 24h,用Annexin V FITC/PI流式细胞术检测HSC凋亡。结果:姜黄素浓度为10μmol/L时,早期凋亡细胞开始...目的:探讨Annexin V FITC/PI流式细胞术在检测姜黄素诱导的肝星状细胞(HSC)凋亡中的应用。方法:不同浓度姜黄素作用肝星状细胞株HSC-T6 24h,用Annexin V FITC/PI流式细胞术检测HSC凋亡。结果:姜黄素浓度为10μmol/L时,早期凋亡细胞开始增多,至30μmol/L时,坏死细胞开始增多,随姜黄素浓度增高,各组凋亡细胞随之增多。0、10、20、30、40μmol/L5组的凋亡率分别是(3.81±0.64)%、(6.52±1.88)%、(11.61±2.79)%、(52.03±4.38)%、(87.11±12.62)%,与0μmol/L组比较,20、30、40μmol/L3组的凋亡率差异有统计学意义(P<0.05)。结论:Annexin V FITC/PI流式细胞术可定量检测姜黄素诱导的HSC早期凋亡。展开更多
To quantitatively analyze apoptotic and secondary necrotic cells unde r apoptosis conditions. Methods. The cells of Burkitt lymphoma (BL) cell line Raji were incubated with 1 .0 μmol/L dexamethasone (DEX) for 2, 4 an...To quantitatively analyze apoptotic and secondary necrotic cells unde r apoptosis conditions. Methods. The cells of Burkitt lymphoma (BL) cell line Raji were incubated with 1 .0 μmol/L dexamethasone (DEX) for 2, 4 and 8 h respectively, then stained with Annexin V FITC (fluorescein isothiocyanate conjugated) which was used to detec t the exposed phosphatidylserine (PS) on the epimembrane resulting from a loss o f phospholipid asymmetry in the early stage of apoptosis, and also stained with propidium iodide (PI) which allows analysis of secondary necrotic cells related with cell membrane and DNA damage that probably represent late stage of apoptosi s, then apoptotic cells were quantified by flow cytometry (FCM). Furthermore, An nexin+/PI and Annexin+/PI+ cells were sorted by fluoresence activated cell sorter (FACS), and identified by electron microscopy (EM) and DNA gel electroph oresis. Results. The percentage of apoptotic cells was found to increase with the incuba tion time (r=0.97). This method was sensitive with low detection limit (0.02%), and was reproducible with low coefficient variance (CV)(4.2%). Meanwhile, the Annexin+/PI and Annexin+/PI+ cells were identified as apoptotic and necroti c cells under EM, and DNA extracted from the Annexin+/PI cells was characteri zed by “ladder pattern”. Conclusions. Annexin V assay is a specific, sensitive, accurate, reproductive and quantitative method for analyzing apoptotic cells.展开更多
文摘目的:探讨Annexin V FITC/PI流式细胞术在检测姜黄素诱导的肝星状细胞(HSC)凋亡中的应用。方法:不同浓度姜黄素作用肝星状细胞株HSC-T6 24h,用Annexin V FITC/PI流式细胞术检测HSC凋亡。结果:姜黄素浓度为10μmol/L时,早期凋亡细胞开始增多,至30μmol/L时,坏死细胞开始增多,随姜黄素浓度增高,各组凋亡细胞随之增多。0、10、20、30、40μmol/L5组的凋亡率分别是(3.81±0.64)%、(6.52±1.88)%、(11.61±2.79)%、(52.03±4.38)%、(87.11±12.62)%,与0μmol/L组比较,20、30、40μmol/L3组的凋亡率差异有统计学意义(P<0.05)。结论:Annexin V FITC/PI流式细胞术可定量检测姜黄素诱导的HSC早期凋亡。
文摘To quantitatively analyze apoptotic and secondary necrotic cells unde r apoptosis conditions. Methods. The cells of Burkitt lymphoma (BL) cell line Raji were incubated with 1 .0 μmol/L dexamethasone (DEX) for 2, 4 and 8 h respectively, then stained with Annexin V FITC (fluorescein isothiocyanate conjugated) which was used to detec t the exposed phosphatidylserine (PS) on the epimembrane resulting from a loss o f phospholipid asymmetry in the early stage of apoptosis, and also stained with propidium iodide (PI) which allows analysis of secondary necrotic cells related with cell membrane and DNA damage that probably represent late stage of apoptosi s, then apoptotic cells were quantified by flow cytometry (FCM). Furthermore, An nexin+/PI and Annexin+/PI+ cells were sorted by fluoresence activated cell sorter (FACS), and identified by electron microscopy (EM) and DNA gel electroph oresis. Results. The percentage of apoptotic cells was found to increase with the incuba tion time (r=0.97). This method was sensitive with low detection limit (0.02%), and was reproducible with low coefficient variance (CV)(4.2%). Meanwhile, the Annexin+/PI and Annexin+/PI+ cells were identified as apoptotic and necroti c cells under EM, and DNA extracted from the Annexin+/PI cells was characteri zed by “ladder pattern”. Conclusions. Annexin V assay is a specific, sensitive, accurate, reproductive and quantitative method for analyzing apoptotic cells.
