Gerasimenko Julia V, Flowerdew Sarah E, Voronina Svetlana G, Sukhomlin Tatiana K, Tepikin Alexei V, Petersen Ole H, Gerasimenko Oleg V
MRC Secretory Control Research Group, The Physiological Laboratory, University of Liverpool, Crown Street, Liverpool, L69 3BX United Kingdom.
J Biol Chem. 2006 Dec 29;281(52):40154-63. doi: 10.1074/jbc.M606402200. Epub 2006 Oct 30.
Gallstones can cause acute pancreatitis, an often fatal disease in which the pancreas digests itself. This is probably because of biliary reflux into the pancreatic duct and subsequent bile acid action on the acinar cells. Because Ca(2+) toxicity is important for the cellular damage in pancreatitis, we have studied the mechanisms by which the bile acid taurolithocholic acid 3-sulfate (TLC-S) liberates Ca(2+). Using two-photon plasma membrane permeabilization and measurement of [Ca(2+)] inside intracellular stores at the cell base (dominated by ER) and near the apex (dominated by secretory granules), we have characterized the Ca(2+) release pathways. Inhibition of inositol trisphosphate receptors (IP(3)Rs), by caffeine and 2-APB, reduced Ca(2+) release from both the ER and an acidic pool in the granular area. Inhibition of ryanodine receptors (RyRs) by ruthenium red (RR) also reduced TLC-S induced liberation from both stores. Combined inhibition of IP(3)Rs and RyRs abolished Ca(2+) release. RyR activation depends on receptors for nicotinic acid adenine dinucleotide phosphate (NAADP), because inactivation by a high NAADP concentration inhibited release from both stores, whereas a cyclic ADPR-ribose antagonist had no effect. Bile acid-elicited intracellular Ca(2+) liberation from both the ER and the apical acidic stores depends on both RyRs and IP(3)Rs.
胆结石可引发急性胰腺炎,这是一种常具致命性的疾病,患病时胰腺会自我消化。这可能是由于胆汁反流至胰管,随后胆汁酸作用于腺泡细胞所致。鉴于钙离子毒性在胰腺炎细胞损伤中至关重要,我们研究了胆汁酸牛磺石胆酸3 - 硫酸盐(TLC - S)释放钙离子的机制。利用双光子质膜通透化技术以及测量细胞基部(以内质网为主)和顶端附近(以分泌颗粒为主)细胞内储存库中的钙离子浓度,我们对钙离子释放途径进行了表征。咖啡因和2 - 氨基乙氧基二苯硼酸(2 - APB)抑制肌醇三磷酸受体(IP₃Rs),减少了内质网和颗粒区域酸性池中的钙离子释放。钌红(RR)抑制兰尼碱受体(RyRs)也减少了TLC - S诱导的两个储存库中的钙离子释放。联合抑制IP₃Rs和RyRs可消除钙离子释放。RyR的激活依赖于烟酰胺腺嘌呤二核苷酸磷酸(NAADP)受体,因为高浓度NAADP使其失活会抑制两个储存库中的钙离子释放,而环ADP - 核糖拮抗剂则无作用。胆汁酸引发的内质网和顶端酸性储存库中的细胞内钙离子释放依赖于RyRs和IP₃Rs。