摘要
将草鱼肾细胞(CIK)在28℃条件下培养24h后,分别置于28(对照)、32、36和40℃的条件下暴露2h,测定不同温度处理组的细胞乳酸脱氢酶(LDH)漏出率、细胞周期的改变、细胞内多泛素化蛋白水平以及caspase-3和bcl-2的表达变化;置于28(对照)、32、36和40℃的条件下暴露12h,测定不同温度处理组的细胞活力。试验结果表明,热应激引起细胞活力显著下降,导致CIK细胞的LDH漏出率显著上升,多泛素化蛋白表达量显著增加,bcl-2的表达量显著下调和caspase-3的表达量显著上调,G0/G1期细胞显著减少。急性热应激可能会引起CIK的细胞膜通透性和蛋白质表达量的变化,从而诱导细胞启动凋亡。
The kidney cells of grass carp （Ctenopharyngodon idellus） were exposed to 28 ℃ ,32 ℃, 36 ℃ and 40 ℃ for 2 h or 12 h,respectively. The 28 ℃ group was taken as control. LDH leakage rates of CIK cells were determined by the trace detection of LDH kit. The cell cycles were measured by flow cy- tometry. The polyubiquitinylated protein was measured by dot blot, and the expressions of caspase^3 and bcl-2 genes of cells were detected by real-time fluorescence quantitative PCR. The results showed that the LDH leakage rate, the content of polyubiquitinylated proteins, and the caspase- 3 expression increased significantly with the increase of incubation temperature. While the expression of bcl-2 and the cells at G0/G1 phase decreased significantly with the increasing incubation temperature. Acute thermal stress could induce the damage of cell membrane, protein,and even promote cell apoptosis.
出处
《华中农业大学学报》
CAS
CSCD
北大核心
2014年第4期88-92,共5页
Journal of Huazhong Agricultural University
基金
国家大宗淡水鱼类产业技术体系资金项目(CARS-46-15)
国家自然科学基金项目(30970529)
中央高校基本科研业务费专项(2013PY077)
关键词
急性热应激
草鱼肾细胞
细胞凋亡
acute thermal stress
Ctenopharyngodon idellus kidney cell
apoptosis
