论文部分内容阅读
Objective To study of myocardial cell apoptosis cytomics technology was used, including high-content screening (HCS), flow cytometry (FCM), laser scanning cytometry (LSC).Methods To establish the model of myocardial cell apoptosis in vitro, H9C2 cell line was treated with hypoxia buffer for 6 hours.The DMEM treatment was as control group.The cells were stained with Annexin V-FITC and PI, and then analyzed for the ratio of early and late apoptosis and necrosis using HCS and FCM.Myocardial infraction was performed in mice by ligation of left anterior descending branch.After 28 days, heart sections were performed with TUNEL assay.The myocardium apoptosis was evaluated with HCS and LSC in the normal, border and infracted area of the hearts.Results After 6 hours of hypoxia buffer treatment, HCS method displayed the ratio of early apoptotic cells (Anenxin V+PI-) significantly higher in hypoxia group than DMEM group [(17.69 ± 2.85) vs (5.31 ± 0.86)%, P<0.01], but there was no difference in the ratio of late apoptotic (Anenxin V+PI+) and necrotic (Anenxin V-PI+) cell between the two groups.Moreover, FCM methods revealed the same trend in cell apoptosis.The percentage of early apoptosis significantly increased in hypoxia group compared in DMEM group [(12.18 ± 0.61) vs (3.08 ± 0.15)%, P <0.01].There was no difference in the ratio of late apoptotic and necrotic cells.After 28 days of myocardial infarction, both HCS and LSC methods showed that there were apoptotic cells in the border and infracted zone of the hearts.The percentage of the apoptotic cells were 6.08% (HCS) and 4.35% (LSC) respectively.Conclusions HCS and FCM methods are reliable, stable and consistent in assessment of myocardial cell apoptosis.Two methods are able to quantify and distinguish the process of the apoptosis.LSC and HCS methods are applicable at the tissue sections.LSC may clearly localized apoptotic cells in tissues.