用流式细胞术检测喹乙醇诱导的鲤肝细胞凋亡

Apoptosis of hepatic cell in common carp poisoned with olaquindox by flow cytometry(FCM)

采用含200 mg.kg-1喹乙醇的饲料连续饲喂鲤鱼(Cyprinus carpioLinnaeus),分别于饲喂后的第7天、14天、20天、25天和第30天各剖杀4尾鱼,取肝组织,用流式细胞仪检测其细胞凋亡程度。结果显示,在肝细胞DNA直方图中,于G1峰之前出现一AP峰,且AP峰峰值随染毒时间的延长而升高,肝细胞在不同时期的凋亡率与染毒时间呈正相关。在经喹乙醇染毒第7天,鲤肝细胞的凋亡率平均值为2.05%,而对照组为1.70%,两组间差异不显著(P>0.05);在第14天,可见轻微的AP峰;在第20天,凋亡率达到9.49%,凋亡峰已较明显;在第25天时的AP峰比第14天时峰值明显升高;在染毒第30天时,凋亡率达17.44%,AP峰明显可见,其肝细胞凋亡率与第7天、第14天和第25天时差异显著。在整个过程中,对照组的正常G1峰前几乎看不出有AP峰存在。在实验组内,不同染毒时间,实验各组的肝细胞凋亡率差异显著,对照组内差异不显著。

The fish was fed with food included 200 mg/kg diet olaquindox. Four fish were dissected each time to observe the apoptosis of the hepatic cell by flow cytometry on 7th day, 14th day, 20th day, 25th day and 30th day after being poisoned. The results showed that olaquindox could induce the apoptosis of the hepatic cell in common carp. The AP peaks before the G1 peaks were shown in DNA graphs. Furthermore, the percentage of the apoptosis increased with the time delayed. At the same time, the percentage of carp hepatic cell apoptosis was got using flow cytometry and it was positive-correlation to the time of being poisoned. The average percentage of hepatic cell apoptosis was 2.05% after the carp was poisoned for 7 days. However, it was 1.70% in the control. The difference was not significant （P 〉0.05）, so the AP peak was not distinct yet. A slight AP peak on the 14th day was got and it was clear on the 20th day, and the percentage of apoptosis reached 9.49 %. The AP peak was much higher on the 25th day than on the 14th day. And the percentage of apoptosis reached 17.44% after being poisoned for 30 days, which was significant compared with those on 7th, 14th and 25th days. During the whole period, the AP peak before the G1 peak in the control was not seen. Data analysis showed that the difference of the hepatic cell apoptosis was not distinct in the control. And the difference of hepatic apoptosis percentage was distinct in different time in the trial. However, it was not distinct in the control.