Calcium-dependent ultrasound stimulation of secretory events from pancreatic beta cells
Calcium-dependent ultrasound stimulation of secretory events from pancreatic beta cells
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DOI：
10.1186/s40349-017-0108-9
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发表时间：
2017-12-05
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影响因子：
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通讯作者：
Zderic, Vesna
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文献类型：
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作者：
Castellanos, Ivan Suarez;Singh, Tania;Zderic, Vesna
背景资料：我们以前的研究表明，超声可以刺激胰腺β细胞释放胰岛素，为2型糖尿病提供了一种潜在的新治疗方法。本研究的目的是探讨时间动力学和钙离子依赖性的超声刺激分泌事件从多巴胺加载的胰腺β细胞在体外setting.Methods：碳纤维安培法检测分泌从INS-1832/13 β细胞在真实的时间。还使用胰岛素特异性ELISA试剂盒测量响应于超声处理的释放的胰岛素水平。将β细胞暴露于强度为0.1 W/cm（2）、0.5W/cm（2）和1 W/cm（2）的800 kHz连续波超声数秒。结果：碳纤维安培法实验表明，800 kHz、强度为0.5和1 W/cm（2）的超声波能够刺激细胞分泌，刺激持续时间与刺激持续时间相同。此外，在暴露于0.5 W/cm（2）超声强度的细胞中，当细胞外Ca 2+与10 mM EGTA螯合时，检测到的峰的幅度降低了64%（p < 0.01）。对响应于超声刺激而释放的胰岛素的测量显示通过用10 mM EGTA螯合细胞外Ca 2+完全抑制胰岛素分泌（p < 0.01）。活力研究表明，800 kHz，0.5 W/cm（2）的超声波并没有造成任何显着影响的细胞暴露于超声波的活力和代谢活动相比，假处理cells.Conclusions：我们的研究结果表明，应用超声波是能够刺激胰岛素从胰腺β细胞在一个安全的，可控的和Ca 2+依赖的方式释放。
Background: Our previous studies have indicated that ultrasound can stimulate the release of insulin from pancreatic beta cells, providing a potential novel treatment for type 2 diabetes. The purpose of this study was to explore the temporal dynamics and Ca2+-dependency of ultrasound-stimulated secretory events from dopamineloaded pancreatic beta cells in an in vitro setup.Methods: Carbon fiber amperometry was used to detect secretion from INS-1832/13 beta cells in real time. The levels of released insulin were also measured in response to ultrasound treatment using insulin-specific ELISA kit. Beta cells were exposed to continuous wave 800 kHz ultrasound at intensities of 0.1 W/cm(2), 0.5W/cm(2) and 1 W/cm(2) for several seconds. Cell viability tests were done with trypan blue dye exclusion test and MTT analysis.Results: Carbon fiber amperometry experiments showed that application of 800 kHz ultrasound at intensities of 0.5 and 1 W/cm(2) was capable of stimulating secretory events for durations lasting as long as the duration of the stimulus. Furthermore, the amplitude of the detected peaks was reduced by 64% (p < 0.01) when extracellular Ca2+ was chelated with 10 mM EGTA in cells exposed to ultrasound intensity of 0.5 W/cm(2). Measurements of released insulin in response to ultrasound stimulation showed complete inhibition of insulin secretion by chelating extracellular Ca2+ with 10 mM EGTA (p < 0.01). Viability studies showed that 800 kHz, 0.5 W/cm(2) ultrasound did not cause any significant effects on viability and metabolic activity in cells exposed to ultrasound as compared to sham-treated cells.Conclusions: Our results demonstrated that application of ultrasound was capable of stimulating the release of insulin from pancreatic beta cells in a safe, controlled and Ca2+-dependent manner.