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离体大鼠胰腺切片超声刺激代谢活性的影像学研究。

Ex Vivo Imaging of Ultrasound-Stimulated Metabolic Activity in Rat Pancreatic Slices.

机构信息

Department of Biomedical Engineering, The George Washington University, Washington, DC, USA.

Department of Biological Sciences, George Washington University, Washington, DC, USA.

出版信息

Ultrasound Med Biol. 2021 Mar;47(3):666-678. doi: 10.1016/j.ultrasmedbio.2020.10.021. Epub 2020 Nov 27.

Abstract

Ultrasound has previously been reported to produce a reversible stimulatory effect in cultured rat beta cells. Here, we quantified and assessed dynamic metabolic changes in an in situ pancreatic slice model evoked by ultrasound application. After plating, pancreas slices were imaged using a confocal microscope at 488 and 633 nm to image lipodamine dehydrogenase (Lip-DH) autofluorescence and a far red fluorescence, respectively. Ultrasound was applied at intensities of 0.5 and 1 W/cm at both 800 kHz and 1 MHz. Additionally, 800 kHz at 1 W/cm was applied in a pulsing scheme. No ultrasound (control) and glucose application experiments were performed. A difference in fluorescence signal before and after treatment application was the metric for analysis. Comparison of experimental groups using far red fluorescence revealed significant differences between all experimental groups and control in the islet (p < 0.05) and between all ultrasound experimental groups and control (p < 0.05) in pancreatic exocrine tissue. However, this difference in response between control and glucose did not exist in the exocrine tissue. We also observed using Lip-DH autofluorescence that glucose produces a significantly increased metabolic response in islet tissue compared with exocrine tissue (p < 0.05). Pulsed ultrasound appeared to increase metabolic activity in the pancreatic slice in a more consistent manner compared with continuous ultrasound application. Our results indicate that therapeutic ultrasound may have a stimulatory metabolic effect on the pancreatic islets similar to that of glucose.

摘要

超声先前被报道在培养的大鼠β细胞中产生可逆转的刺激作用。在这里,我们在原位胰腺切片模型中定量和评估了超声应用引起的动态代谢变化。在接种后,使用共聚焦显微镜在 488 和 633nm 处对胰腺切片进行成像,分别对脂肪酶脱氢酶(Lip-DH)自发荧光和远红荧光进行成像。在 800kHz 和 1MHz 时,分别以 0.5 和 1W/cm 的强度施加超声。此外,以 1W/cm 的 800kHz 施加脉冲方案。进行无超声(对照)和葡萄糖应用实验。处理应用前后荧光信号的差异是分析的指标。使用远红荧光比较实验组,在胰岛(p<0.05)和胰腺外分泌组织中所有超声实验组与对照组之间(p<0.05)均发现所有实验组与对照组之间存在显著差异。然而,在外分泌组织中,控制组和葡萄糖组之间的这种反应差异不存在。我们还观察到使用 Lip-DH 自发荧光,与外分泌组织相比,葡萄糖使胰岛组织产生明显增加的代谢反应(p<0.05)。与连续超声应用相比,脉冲超声似乎以更一致的方式增加胰腺切片的代谢活性。我们的结果表明,治疗性超声可能对胰岛具有类似于葡萄糖的刺激代谢作用。

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