Jiang Jingjing, Zhou Zongguang, Wang Ling, Chen Lihui, Li Yuan, Yan Hui, Zhou Bin, Liu Yong, Chen Keling
Institute of Digestive Surgery, West China Hospital, Sichuan University, Chengdu 610041, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2011 Apr;28(2):332-6, 351.
To study the expression of X-linked inhibitor of apoptosis protein (XIAP) and cell apoptosis in vitro model of acute pancreatitis (AP), we carried out experiments to stimulate AR42J cell line with caerulein (10(-8) mol/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-kappaB) was measured by flow cytometry (FCM). With the stimulation of caerulein, the expression of XIAP and the NF-kappaB 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-kappaB can upgrade the expression of XIAP and increase the cell apoptosis.
为研究X连锁凋亡抑制蛋白（XIAP）的表达及细胞凋亡在急性胰腺炎（AP）体外模型中的情况，我们进行实验，用雨蛙素（10⁻⁸ mol/L）刺激AR42J细胞系12小时，然后在不同时间点（分别为0小时、4小时、8小时、12小时和24小时）收集细胞。接着，我们用电子显微镜观察经雨蛙素刺激后AR42J细胞的形态变化。采用实时荧光定量聚合酶链反应（FQ-PCR）检测XIAP、半胱天冬酶-3（caspase-3）和半胱天冬酶-9（caspase-9）的基因表达，并用蛋白质印迹法评估XIAP的蛋白质表达。通过流式细胞术（FCM）检测核因子-κB（NF-κB）的激活情况。经雨蛙素刺激后，XIAP的表达及NF-κB的激活先降低后升高，但caspase-3和caspase-9表达的变化则相反。在雨蛙素刺激下，XIAP可能最初抑制大鼠胰腺腺泡AR42J细胞系中的细胞凋亡，随后NF-κB可上调XIAP的表达并增加细胞凋亡。
