Human Pancreatic Acinar Cells Proteomic Characterization, Physiologic Responses, and Organellar Disorders in ex Vivo Pancreatitis
Human Pancreatic Acinar Cells Proteomic Characterization, Physiologic Responses, and Organellar Disorders in ex Vivo Pancreatitis
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DOI：
10.1016/j.ajpath.2017.08.017
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发表时间：
2017-12-01
影响因子：
6
通讯作者：
Pandol, Stephen J.
中科院分区：
文献类型：
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作者：
Lugea, Aurelia;Waldron, Richard T.;Pandol, Stephen J.
对急性胰腺炎分子机制的了解很大程度上是基于对啮齿动物的研究。为了评估在人类中类似的机制，我们在从器官捐赠者身体胰腺分离的人腺泡中进行了体外胰腺炎研究。由于关于这些人的腺泡制剂的数据很少，我们使用光、荧光和电子显微镜评估了它们的功能完整性以及细胞和细胞器的形态，并用液相色谱串联质谱仪评估了它们的蛋白质组。还分析了腺泡细胞对M胆碱激动剂卡巴胆碱(CCH)和胆汁酸3-硫酸盐的反应。来自不同种族的捐赠者的腺泡蛋白组学分析显示，参与翻译、分泌和内溶体功能的消化酶和蛋白质的图谱相似。人腺泡细胞优先表达M受体M3，并对CCH维持至少20小时的生理反应。与啮齿动物腺泡一样，暴露于毒性浓度的CCH和牛磺酸胆酸3-硫酸盐的人腺泡以胰蛋白酶原激活、细胞活力降低、细胞器损伤表现为线粒体去极化、自噬紊乱和病理性内质网应激而反应。急性胰腺炎患者分泌炎性介质，包括IL-6、肿瘤坏死因子-α、IL-1β、趋化因子(C-C基序)配体2和3、巨噬细胞抑制因子以及介导中性粒细胞和单核细胞的趋化因子。总之，人的身体胰腺腺泡保持了生理功能，并具有与啮齿动物腺泡类似的引起胰腺炎的治疗的病理反应和器官紊乱。
Knowledge of the molecular mechanisms of acute pancreatitis is Largely based on studies using rodents. To assess similar mechanisms in humans, we performed ex vivo pancreatitis studies in human acini isolated from cadaveric pancreata from organ donors. Because data on these human acinar preparationS are sparse, we assessed their functional integrity and cellular and organellar morphology using Light, fluorescence, and electron microscopy; and their proteome by Liquid chromatography tandem mass spectrometry. Acinar cell responses to the muscarinic agonist carbachol (CCh) and the bile acid taurolithocholic acid 3-sulfate were also analyzed. Proteomic analysis of acini from donors of diverse ethnicity showed similar profiles of digestive enzymes and proteins involved in translation, secretion, and endolysosomal function. Human acini preferentially expressed the muscarinic acetylcholine receptor M3 and maintained physiological responses to CCh for at least 20 hours. As in rodent acini, human acini exposed to toxic concentrations of CCh and taurolithocholic acid 3-sulfate responded with trypsinogen activation, decreased cell viability, organelle damage manifest by mitochondriaL depolarization, disordered autophagy, and pathological endoplasmic reticulum stress. Human acini also secreted inflammatory mediators elevated in acute pancreatitis patients, including IL-6, tumor necrosis factor-alpha, IL-1 beta, chemokine (C-C motif) ligands 2 and 3, macrophage inhibitory factor, and chemokines mediating neutrophil and monocyte infiltration. In conclusion, human cadaveric pancreatic acini maintain physiological functions and have similar pathological responses and organellar disorders with pancreatitis-causing treatments as observed in rodent acini.