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Ex Vivo Imaging of Ultrasound-Stimulated Metabolic Activity in Rat Pancreatic Slices
Ultrasound in Medicine & Biology ( IF 2.4 ) Pub Date : 2020-11-27 , DOI: 10.1016/j.ultrasmedbio.2020.10.021
Andrew W Chen 1 , Aleksandar Jeremic 2 , Vesna Zderic 1
Affiliation  

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/cm2 at both 800 kHz and 1 MHz. Additionally, 800 kHz at 1 W/cm2 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.



中文翻译:


大鼠胰腺切片超声刺激代谢活动的离体成像



此前曾有报道称超声波对培养的大鼠β细胞产生可逆的刺激作用。在这里,我们量化和评估了超声应用引起的原位胰腺切片模型中的动态代谢变化。铺板后,使用共焦显微镜在 488 和 633 nm 处对胰腺切片进行成像,分别对脂达明脱氢酶 (Lip-DH) 自发荧光和远红荧光进行成像。在800kHz和1MHz下以0.5和1W/cm 2的强度施加超声波。此外,在脉冲方案中应用了1 W/cm 2的800 kHz。没有进行超声(对照)和葡萄糖应用实验。处理应用之前和之后荧光信号的差异是分析的指标。使用远红荧光对实验组进行比较显示,所有实验组与对照组之间的胰岛差异显着( p < 0.05),所有超声实验组与对照组之间的胰腺外分泌组织也存在显着差异( p < 0.05)。然而,外分泌组织中不存在对照和葡萄糖之间的这种反应差异。我们还使用 Lip-DH 自发荧光观察到,与外分泌组织相比,葡萄糖在胰岛组织中产生显着增加的代谢反应( 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中的代谢活动。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。

更新日期:2021-01-15
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