Departments of Pediatrics.
Departments of Internal Medicine, Children's Hospital of Pittsburgh of UPMC, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15224.
J Biol Chem. 2013 Sep 20;288(38):27128-27137. doi: 10.1074/jbc.M113.494534. Epub 2013 Aug 12.
Aberrant Ca(2+) signals within pancreatic acinar cells are an early and critical feature in acute pancreatitis, yet it is unclear how these signals are generated. An important mediator of the aberrant Ca(2+) signals due to bile acid exposure is the intracellular Ca(2+) channel ryanodine receptor. One putative activator of the ryanodine receptor is the nucleotide second messenger cyclic ADP-ribose (cADPR), which is generated by an ectoenzyme ADP-ribosyl cyclase, CD38. In this study, we examined the role of CD38 and cADPR in acinar cell Ca(2+) signals and acinar injury due to bile acids using pharmacologic inhibitors of CD38 and cADPR as well as mice deficient in Cd38 (Cd38(-/-)). Cytosolic Ca(2+) signals were imaged using live time-lapse confocal microscopy in freshly isolated mouse acinar cells during perifusion with the bile acid taurolithocholic acid 3-sulfate (TLCS; 500 μM). To focus on intracellular Ca(2+) release and to specifically exclude Ca(2+) influx, cells were perifused in Ca(2+)-free medium. Cell injury was assessed by lactate dehydrogenase leakage and propidium iodide uptake. Pretreatment with either nicotinamide (20 mM) or the cADPR antagonist 8-Br-cADPR (30 μM) abrogated TLCS-induced Ca(2+) signals and cell injury. TLCS-induced Ca(2+) release and cell injury were reduced by 30 and 95%, respectively, in Cd38-deficient acinar cells compared with wild-type cells (p < 0.05). Cd38-deficient mice were protected against a model of bile acid infusion pancreatitis. In summary, these data indicate that CD38-cADPR mediates bile acid-induced pancreatitis and acinar cell injury through aberrant intracellular Ca(2+) signaling.
胰腺腺泡细胞内异常的 Ca(2+)信号是急性胰腺炎的早期和关键特征,但目前尚不清楚这些信号是如何产生的。在胆汁酸暴露引起的异常 Ca(2+)信号中,一种重要的介质是细胞内 Ca(2+)通道ryanodine 受体。ryanodine 受体的一种假定激活剂是核苷酸第二信使环 ADP-核糖(cADPR),它是由细胞外酶 ADP-核糖基环化酶 CD38 产生的。在这项研究中,我们使用 CD38 和 cADPR 的药理学抑制剂以及缺乏 Cd38(Cd38(-/-))的小鼠,研究了 CD38 和 cADPR 在胆汁酸引起的腺泡细胞 Ca(2+)信号和腺泡损伤中的作用。在新鲜分离的小鼠腺泡细胞中,使用活细胞延时共聚焦显微镜,在胆汁酸牛磺胆酸 3-硫酸盐(TLCS;500 μM)灌流期间,对细胞内 Ca(2+)信号进行成像。为了关注细胞内 Ca(2+)释放,并专门排除 Ca(2+)内流,细胞在无 Ca(2+)的培养基中进行灌流。通过乳酸脱氢酶漏出和碘化丙啶摄取来评估细胞损伤。用烟酰胺(20 mM)或 cADPR 拮抗剂 8-Br-cADPR(30 μM)预处理可阻断 TLCS 诱导的 Ca(2+)信号和细胞损伤。与野生型细胞相比,Cd38 缺陷型腺泡细胞中 TLCS 诱导的 Ca(2+)释放和细胞损伤分别减少了 30%和 95%(p < 0.05)。Cd38 缺陷型小鼠对胆汁酸输注性胰腺炎模型具有保护作用。总之,这些数据表明 CD38-cADPR 通过异常的细胞内 Ca(2+)信号转导介导胆汁酸诱导的胰腺炎和腺泡细胞损伤。