【目的】通过磷脂结合蛋白V/碘化丙啶(Annexin V/PI)流式细胞分析法和原位末端脱氧核苷酸转移酶标记(TUNEL)法检测低氧胁迫下鲢肝、脑细胞的凋亡情况。【方法】将鲢分成对照组和低氧处理组,对照组正常饲养,低氧处理组用保鲜膜将水箱上口封住进行低氧处理,分别在鲢浮头期以及死亡期取其肝、脑组织,同时取对照组鲢肝、脑组织,采用Annexin V/PI流式细胞分析法和TUNEL法检测其细胞凋亡情况。【结果】Annexin V/PI流式细胞仪分析结果表明,在鲢肝脏中细胞凋亡率依次是对照组8.85%、低氧处理组浮头期22.06%、低氧处理组死亡期22.91%;脑中细胞凋亡率依次是对照组8.18%、低氧处理组浮头期24.80%、低氧处理组死亡期24.86%。TUNEL法检测结果显示,肝脏中细胞凋亡率是对照组3.80%、低氧处理组浮头期36.78%、低氧处理组死亡期37.27%;脑中细胞凋亡率依次是对照组6.74%、低氧处理组浮头期35.04%、低氧处理组死亡期35.71%。与正常对照组比较,低氧胁迫后鲢肝、脑细胞凋亡率极显著增大,但随着低氧胁迫时间延长细胞凋亡率无明显差异。【结论】低氧胁迫会极显著促进鲢肝和脑细胞发生凋亡,但鲢肝、脑组织中细胞凋亡率不会随着低氧胁迫时间延长而产生显著差异。
[Objective] The apoptosis of hepatocyte and brain cells of silver carp(Hypophthalmichthys molitrix) under hypoxic stress was investigated by Annexin V/PI with flow cytometry and TUNEL. [Method] Silver carp individuals were divided intonormal control group and hypoxic stress treatment group. The normal control group was normally bred while the hypoxic stress treatment group was treated with hypoxia in water tank, which was sealed with plastic wrap. Samples of liver and brain of silver carp were collected from normal and treatment groups at floating and death stages to analyze the apoptosis of hepatocyte and brain cells using Annexin V/PI with flow cytometry and TUNEL. [Result] Rates of hepatocyte apoptosis of silver carp were 8.85% for control group,22.06% at floating stage and 22.91%at death stage. Rates of brain cells apoptosis were 8.18% ,24.80% and 24.86% accordingly. TUNEL measurement showed that rates of hepatocyte apoptosis were 3.80% ,36.78% and 37.27% for control,floating state and death stage,while those of brain cells apoptosis were 6. 74%,35.04% and 35. 71%. Compared with the control group,the apoptosis of hepatocyte and brain cells of silver carp was significantly increased under hypoxic stress. However,with the increase of hypoxia time, the number of apoptosis cells did not change significantly. [Conclusion] Hypoxia stress significantly promoted the apoptosis of hepatocyte and brain cells of silver carp and the rates were not significantly changed by hypoxia time.