丙酮酸和谷氨酰胺决定胆囊收缩素和乙醇对大鼠胰腺腺泡线粒体氧化、坏死及形态的影响。

Pyruvate and Glutamine Define the Effects of Cholecystokinin and Ethanol on Mitochondrial Oxidation, Necrosis, and Morphology of Rat Pancreatic Acini.

作者信息

Manko Bohdan O, Bilonoha Olha O, Voloshyn Dariia M, Zub Anastasiia M, Ivasechko Iryna I, Manko Volodymyr V

机构信息

From the Human and Animal Physiology Department, Ivan Franko National University of Lviv.

Institute of Cell Biology of National Academy of Sciences of Ukraine, Lviv, Ukraine.

出版信息

Pancreas. 2021 Aug 1;50(7):972-981. doi: 10.1097/MPA.0000000000001864.

DOI:10.1097/MPA.0000000000001864

PMID:34629447

Abstract

OBJECTIVES

The objective of this study was to test whether pyruvate and glutamine affect the ethanol and cholecystokinin (CCK) effects on the mitochondrial function, viability, and morphology of rat pancreatic acini.

METHODS

Respiration was measured with Clark oxygen electrode. Mitochondrial membrane potential, reduced nicotinamide adenine dinucleotide (phosphate) (NAD(P)H), cell morphology, and viability were studied with fluorescence microscopy.

RESULTS

In vitro, CCK (0.1 nM) caused pyruvate-dependent stimulation of basal and uncoupled respiration, and the effects were abolished by ethanol (20 mM). The combination of ethanol with CCK (2 hours) caused necrosis of approximately 40% acinar cells in medium with glucose, but not with pyruvate and/or glutamine. Cholecystokinin (10 nM) or ethanol with 0.1 nM CCK caused plasma membrane blebbing not related to apoptosis only when both glutamine and pyruvate were present. Glutamine, but not pyruvate, decreased NAD(P)H level and prevented the effects of ethanol with CCK on mitochondrial membrane potential and NAD(P)H, but, in combination with CCK and ethanol, decreased the uncoupled respiration. In vivo, the combination of ethanol (4 g/kg) and CCK (20 pmol/kg) suppressed basal and uncoupled respiration and caused acinar cell blebbing, but not necrosis.

CONCLUSIONS

The lack of sufficient substrate supply in vitro makes pancreatic acinar cells susceptible to necrosis caused by ethanol and CCK in clinically relevant concentrations.

摘要

目的

本研究旨在测试丙酮酸和谷氨酰胺是否会影响乙醇和胆囊收缩素（CCK）对大鼠胰腺腺泡线粒体功能、活力及形态的作用。

方法

使用克拉克氧电极测量呼吸。通过荧光显微镜研究线粒体膜电位、还原型烟酰胺腺嘌呤二核苷酸（磷酸）（NAD(P)H）、细胞形态及活力。

结果

在体外，CCK（0.1 nM）可引起丙酮酸依赖性的基础呼吸和非偶联呼吸刺激，而乙醇（20 mM）可消除这些作用。在含葡萄糖的培养基中，乙醇与CCK联合作用（2小时）可导致约40%的腺泡细胞坏死，但在含丙酮酸和/或谷氨酰胺的培养基中则不会。仅当同时存在谷氨酰胺和丙酮酸时，胆囊收缩素（10 nM）或乙醇与0.1 nM CCK才会引起与凋亡无关的质膜起泡。谷氨酰胺而非丙酮酸可降低NAD(P)H水平，并防止乙醇与CCK对线粒体膜电位和NAD(P)H的影响，但与CCK和乙醇联合使用时，会降低非偶联呼吸。在体内，乙醇（4 g/kg）和CCK（20 pmol/kg）联合作用可抑制基础呼吸和非偶联呼吸，并引起腺泡细胞起泡，但不会导致坏死。