■细胞外囊泡(EV)已从各种来源分离,包括原代和培养的细胞系和体液。以前的研究,包括在我们实验室进行的那些,报道了电动汽车在各种储存条件下的稳定性。
■通过尺寸排阻色谱法分离来自人类全唾液的EV。为了模拟胃液或肠液对电动汽车稳定性的影响,将胃蛋白酶或胰酶添加到样品中。此外,为了确定胆汁酸的作用,加入胆酸钠。然后对样品进行蛋白质印迹,动态光散射,和透射电子显微镜分析。此外,检查保留在样品中的二肽基肽酶(DPP)IV的活性,以监测EV的稳定性.
■在酸性条件下,胃蛋白酶模仿胃的环境,电动汽车保持稳定。然而,在胰酶和胆酸钠的存在下,它们部分失去了膜的完整性,表明它们通过十二指肠后可能不稳定。虽然有几种相关的蛋白质,如黏蛋白5B和CD9被降解,民进党四是稳定的,其活性在模拟胃肠条件下得以保留。
■我们的数据表明,尽管电动汽车可以通过胃而不会受到明显损害,它们可能会在肠道中被破坏以释放其内容物。活性成分如DPPIV从EV持续递送到肠道中可能在胃肠道中发生的信号转导途径的稳态的有效调节中起作用。
UNASSIGNED: Extracellular vesicles (EVs) have been isolated from various sources, including primary and cultured cell lines and body fluids. Previous studies, including those conducted in our laboratory, have reported the stability of EVs under various storage conditions.
UNASSIGNED: EVs from human whole saliva were separated via size-exclusion chromatography. To simulate the effects of gastric or intestinal fluids on the stability of EVs, pepsin or pancreatin was added to the samples. Additionally, to determine the effect of bile acids, sodium cholate was added. The samples were then subjected to western blotting, dynamic light scattering, and transmission electron microscopy analyses. In addition, the activity of dipeptidyl peptidase (DPP) IV retained in the samples was examined to monitor the stability of EVs.
UNASSIGNED: Under acidic conditions, with pepsin mimicking the milieu of the stomach, the EVs remained stable. However, they partially lost their membrane integrity in the presence of pancreatin and sodium cholate, indicating that they may be destabilized after passing through the duodenum. Although several associated proteins, such as mucin 5B and CD9 were degraded, DPP IV was stable, and its activity was retained under the simulated gastrointestinal conditions.
UNASSIGNED: Our data indicate that although EVs can pass through the stomach without undergoing significant damage, they may be disrupted in the intestine to release their contents. The consistent delivery of active components such as DPP IV from EVs into the intestine might play a role in the efficient modulation of homeostasis of the signal transduction pathways occurring in the gastrointestinal tract.