摘要： 拟探讨热应激诱导LLC-PK1细胞线粒体凋亡相关因子的表达和活性情况，以及激活线粒体凋亡通路的时间。采用qRT-PCR方法和Caspase活性检测试剂盒分别检测42℃热应激处理后0，2，4，8，16 h以及37℃培养的LLC-PK1细胞中HSP72、Bcl-2、Bax、AIF和Cyt.c的基因表达以及Caspase-9和Caspase-3的活性。结果显示，42℃热应激1 h，LLC-PK1细胞Bcl-2/Bax（P<0.05）和AIF（P<0.01）基因表达以及Caspase-3（P<0.01）活性从0 h开始升高（P<0.01），到2 h达到最高（P<0.01）；HSP72基因表达和Caspase-9活性0 h即达到最高（P<0.01）；Cyt.c基因表达在2 h开始极显著升高（P<0.01），并且此时最高，8 h以后全部因子恢复到正常水平。结果表明，42℃热应激1 h即激活LLC-PK1细胞线粒体凋亡通路，且热应激后的0～8 h线粒体凋亡通路处于活化状态，其中2 h时最为活跃。

关键词: 热应激, 猪肾小管上皮细胞, 线粒体凋亡

Abstract: To investigate the expression and the activity of factors related to mitochondrial apoptosis in LLC-PK1 cells induced by heat stress,and the time of activation of mitochondrial apoptotic pathway,we assessed the HSP72, Bcl-2, Bax,AIF,Cyt.c expressions in LLC-PK1 cells by qRT-PCR and Caspase-9,Caspase-3 activity by caspase activity assay kit at 0,2,4,8,16 h after 42℃ heat stress treatment for 1 h.The above factors were also measured in cells cultured at 37℃ as control.Results demonstrated that Bcl-2/Bax and AIF gene expressions and Caspase-3 activity significantly increased from 0 h,and reached the highest point at 2 h after heat stress; HSP72 gene expression and Caspase-9 activity reached the highest point at 0 h;Cyt.c gene expression significantly increased and reached the highest point at 2 h,and all the measured factors returned to normal level at 8 h after heat stress.The above results reveal that mitochondrial apoptotic pathway in LLC-PK1 cells is in a state of activation in 0-8 h after 42℃ heat stress for 1 h and is the most active at 2 h.

Key words: Heat stress, LLC-PK1 cells, Mitochondrial apoptosis

中图分类号: 

• Q78