Mistlberger-Reiner Agnes, Sterneder Sonja, Reipert Siegfried, Wolske Sara, Somoza Veronika
Department of Physiological Chemistry, Faculty of Chemistry, University of Vienna, Vienna 1090, Austria.
Core Facility Cell Imaging and Ultrastructure Research, University of Vienna, Vienna 1030, Austria.
ACS Omega. 2023 Jan 5;8(2):2213-2226. doi: 10.1021/acsomega.2c06442. eCollection 2023 Jan 17.
The secretion of extracellular vesicles and particles (EVPs) is an important mechanism of cellular communication. In this work, we demonstrate a functional role of EVPs in mechanisms regulating gastric acid secretion. HGT-1 cells were used as a model system to assess proton secretion. First, in order to prove EVP secretion by HGT-1 cells, EVPs were isolated by size exclusion chromatography and characterized by nanoparticle tracking analysis, Western blot, and cryo transmission electron microscopy. For examination of the potential role of EVPs in proton secretion, HGT-1 cells were treated with pharmacological EV-inhibitors, resulting in a reduction of histamine-induced proton secretion. To demonstrate the functional role of EVPs in the mechanism of proton secretion, EVP-conditioned supernatant was collected after stimulation of HGT-1 cells with histamine, fractionated, and subjected to an activity screening. The results revealed constituents of the HGT-1-derived secretome with an MW of >100 kDa (including EVPs) to modulate proton secretion, while smaller constituents had no effect. Finally, a dose-dependent modulatory effect on proton secretion of HGT-1 cells was demonstrated by isolated HGT-1-derived EVPs. Hence, this study presents first results on the potential function of EVPs as a previously undiscovered mechanism of regulation of gastric acid secretion by parietal cells.
细胞外囊泡和颗粒（EVPs）的分泌是细胞通讯的重要机制。在本研究中，我们证明了EVPs在调节胃酸分泌机制中的功能作用。使用HGT-1细胞作为评估质子分泌的模型系统。首先，为了证明HGT-1细胞分泌EVPs，通过尺寸排阻色谱法分离EVPs，并通过纳米颗粒跟踪分析、蛋白质印迹和冷冻透射电子显微镜对其进行表征。为了研究EVPs在质子分泌中的潜在作用，用药物性EV抑制剂处理HGT-1细胞，导致组胺诱导的质子分泌减少。为了证明EVPs在质子分泌机制中的功能作用，在用组胺刺激HGT-1细胞后收集EVP条件培养基上清液，进行分级分离并进行活性筛选。结果显示，HGT-1来源的分泌蛋白组中分子量>100 kDa的成分（包括EVPs）可调节质子分泌，而较小的成分则无作用。最后，分离得到的HGT-1来源的EVPs对HGT-1细胞的质子分泌具有剂量依赖性调节作用。因此，本研究首次展示了EVPs作为壁细胞胃酸分泌调节的一种先前未被发现的机制的潜在功能。
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