Dept. of Internal Medicine, Univ. of Michigan Medical School, Ann Arbor, 48109, USA.
Am J Physiol Gastrointest Liver Physiol. 2010 Aug;299(2):G400-12. doi: 10.1152/ajpgi.00061.2010. Epub 2010 Jun 3.
Previously, we found that the University of North Carolina cystic fibrosis (UNC-CF) mouse had more severe experimental acute pancreatitis (AP) than wild-type (WT) mice characterized by exuberant pancreatic inflammation and impaired acinar apoptosis. Because exon 10 CFTR gene mutations exhibit different phenotypes in tissues such as the mouse lung, we tested the hypothesis that DeltaF508-CF mice also develop severe AP associated with an antiapoptotic acinar phenotype, which requires indirect effects of the extracellular milieu. We used cerulein hyperstimulation models of AP. More severe pancreatitis occurred in cerulein-injected DeltaF508-CF vs. WT mice based on histological severity (P < 0.01) and greater neutrophil sequestration [P < 0.0001; confirmed by myeloperoxidase activity (P < 0.005)]. In dispersed acini cerulein-evoked necrosis was greater in DeltaF508-CF acini compared with WT (P < 0.05) and in WT acini pretreated with CFTR(inh)-172 compared with vehicle (P < 0.05). Cerulein-injected DeltaF508-CF vs. WT mice had less apoptosis based on poly(ADP-ribose) polymerase (PARP) cleavage (P < 0.005), absent DNA laddering, and reduced terminal deoxynucleotidyltransferase biotin-dUTP nick end labeling (TUNEL) staining (P < 0.005). Unexpectedly, caspase-3 activation was greater in DeltaF508-CF vs. WT acini at baseline (P < 0.05) and during AP (P < 0.0001). Downstream, DeltaF508-CF pancreas overexpressed the X-linked inhibitor of apoptosis compared with WT (P < 0.005). In summary, the DeltaF508-CF mutation, similar to the UNC-CF "null" mutation, causes severe AP characterized by an exuberant inflammatory response and impaired acinar apoptosis. Enhanced acinar necrosis in DeltaF508-CF occurs independently of extracellular milieu and correlates with loss of CFTR-Cl conductance. Although both exon 10 models of CF inhibit acinar apoptosis execution, the DeltaF508-CF mouse differs by increasing apoptosis signaling. Impaired transduction of increased apoptosis signaling in DeltaF508-CF acini may be biologically relevant to the pathogenesis of AP associated with CFTR mutations.
先前,我们发现北卡罗来纳大学囊性纤维化(UNC-CF)小鼠的实验性急性胰腺炎(AP)比野生型(WT)小鼠更为严重,其特征为胰腺炎症旺盛且腺泡细胞凋亡受损。因为 CFTR 基因第 10 外显子的突变在肺部等组织中表现出不同的表型,所以我们提出假设,即 DeltaF508-CF 小鼠也会发生与抗凋亡腺泡表型相关的严重 AP,这需要细胞外环境的间接作用。我们使用了促胰液素刺激的 AP 模型。基于组织学严重程度(P < 0.01)和中性粒细胞募集增加(P < 0.0001;通过髓过氧化物酶活性证实(P < 0.005)),与 WT 相比,促胰液素注射的 DeltaF508-CF 小鼠发生更严重的胰腺炎。与 WT 相比,DeltaF508-CF 腺泡中促胰液素诱导的坏死更大(P < 0.05),并且用 CFTR(inh)-172 预处理的 WT 腺泡与载体相比(P < 0.05)。与 WT 相比,促胰液素注射的 DeltaF508-CF 小鼠的多聚(ADP-核糖)聚合酶(PARP)裂解(P < 0.005)、无 DNA 梯状带和减少的末端脱氧核苷酸转移酶生物素-dUTP 缺口末端标记(TUNEL)染色(P < 0.005)的凋亡较少。出乎意料的是,DeltaF508-CF 与 WT 相比,在基线(P < 0.05)和 AP 期间(P < 0.0001),腺泡中的 caspase-3 激活更大。下游,与 WT 相比,DeltaF508-CF 胰腺过度表达了 X 连锁凋亡抑制剂(P < 0.005)。总之,DeltaF508-CF 突变与 UNC-CF“空”突变类似,可导致以过度炎症反应和腺泡细胞凋亡受损为特征的严重 AP。DeltaF508-CF 中的增强腺泡坏死独立于细胞外环境发生,并与 CFTR-Cl 电导的丧失相关。尽管两种 10 外显子模型均抑制腺泡细胞凋亡执行,但 DeltaF508-CF 小鼠通过增加凋亡信号而有所不同。DeltaF508-CF 腺泡中增加的凋亡信号转导受损可能与 CFTR 突变相关的 AP 发病机制有关。