摘要
探讨雨蛙素（CAE）刺激大鼠胰腺腺泡细胞AR42J后,其X-连锁凋亡抑制蛋白（XIAP）的表达变化规律及与细胞凋亡的关系。CAE刺激组及空白对照组,加药后继续培养12 h设为0 h时间点,分别在0、4、8、12及24 h收集细胞,电镜观察细胞超微结构变化,实时荧光定量PCR法检测XIAP及细胞凋亡指标caspase-3和caspase-9的mRNA表达,用Western blot法检测XIAP蛋白表达,用流式细胞术检测NF-κB激活率。结果发现,CAE刺激后,AR42J细胞凋亡在8 h最明显;XIAP表达随时间呈先降低后恢复的趋势,并在8 h达最低点（与其余各组差异均有统计学意义,P〈0.05）;caspase-3及caspase-9含量则随时间呈先升高后恢复的趋势,且与XIAP含量呈负相关（P〈0.001）;而NF-κB激活增加,并在4~8 h达高峰。由此推测,在CAE刺激所导致的炎症条件下,XIAP负向调控AR42J细胞的凋亡,而随着炎症的进展,NF-κB的激活逐渐增多,上调XIAP的表达,细胞凋亡指标的表达逐渐恢复。
To study the expression of X-linked inhibitor of apoptosis protein（XIAP） and cell apoptosis in virto model of acute pancreatitis（AP）,we carried out experiments to stimulate AR42J cell line with caerulein（10-8mol/L） for 12 hours,then collected cells at various time points（0 h,4 h,8 h,12 h,and 24 h,respectively）.We then observed the morphologic changes of AR42J cells with the stimulation of caerulein with electronic microscope.The gene expression of XIAP,caspase-3 and caspase-9 was detected using real-time fluorescence quantitative polymerase chain reaction（FQ-PCR）,and the protein expression of XIAP was assessed by western blot.The activation of nuclear factor-kappa B（NF-κB） was measured by flow cytometry（FCM）.With the stimulation of caerulein,the expression of XIAP and the NF-κB activation could first decrease and then increase,but the change of caspase-3 and caspase-9 expressions were opposite.XIAP may inhibit the cell apoptosis in rat pancreatic acinus AR42J cell lines at first with the stimulation of caerulein,then NF-κB can upgrade the expression of XIAP and increase the cell apoptosis.
出处
《生物医学工程学杂志》
EI
CAS
CSCD
北大核心
2011年第2期332-336,351,共6页
Journal of Biomedical Engineering
基金
国家自然科学基金资助项目(30830100)
