Possible role of corticosterone in proteolysis, glycolytic, and amino acid metabolism in primary cultured avian myotubes incubated at high-temperature conditions
Possible role of corticosterone in proteolysis, glycolytic, and amino acid metabolism in primary cultured avian myotubes incubated at high-temperature conditions
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DOI：
10.1016/j.domaniend.2021.106608
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发表时间：
2021-02-18
影响因子：
2.1
通讯作者：
Kikusato, Motoi
中科院分区：
文献类型：
--
作者：
Furukawa, Kyohei;Toyomizu, Masaaki;Kikusato, Motoi
过量的糖皮质激素分泌诱导氧化损伤和肌肉蛋白水解，调节糖脂代谢。众所周知，高温（HT）处理可以促进鸡的皮质酮（CORT）分泌、肌肉蛋白水解和线粒体活性氧（mtROS）的产生。本研究研究了CORT对ht处理细胞中蛋白质水解和mtROS产生以及葡萄糖、脂肪酸和氨基酸代谢的影响。从5只0日龄或1日龄雏鸡胸大肌中分离成肌细胞，在37℃/CO2条件下预培养48 h，达到亚融合（80%-90%）状态。然后将细胞重新接种到6孔或24孔微孔板上进行后续测量，然后在控制温度（37℃，对照）或高温（41℃）条件下培养1或6小时。高温处理的细胞与生理相关浓度的CORT （20 ng/mL二甲亚砜）共培养。与对照组相比，HT处理降低了细胞蛋白含量（P < 0.05），增加了atrogin-1 mRNA水平和mtROS生成水平（P < 0.05），而HT/CORT共处理未引起这两项参数的变化。与正常细胞相比，ht处理的细胞中葡萄糖转运体-1 mRNA水平降低（P < 0.05）， CORT联合处理的细胞中葡萄糖转运体-1 mRNA水平降低（P < 0.05）。虽然HT处理没有改变丙酮酸脱氢酶激酶-4 mRNA的水平，但与对照组和HT处理的细胞相比，CORT联合处理的水平升高（P < 0.05）。HT和HT/CORT处理均未改变脂肪酸氧化相关因子、肉碱棕榈酰转移酶-1和分化簇36的mRNA水平。本研究采用气相色谱-质谱联用进行代谢分析。结果表明，与HT处理的细胞相比，HT/ cort处理的细胞内柠檬酸盐和α -酮戊二酸水平降低（P < 0.05），细胞外具有糖异生特性的丙氨酸和氨基酸水平升高，天冬氨酸、异亮氨酸、丝氨酸、蛋氨酸和苏氨酸水平升高（P < 0.05）。这些结果表明，在高温处理的鸡中，CORT可能不影响蛋白质水解和mtROS的产生，但可以抑制丙酮酸氧化并促进丙氨酸的产生。(C) 2021爱思唯尔公司版权所有。
Excess glucocorticoid secretion induces oxidative damage and muscle proteolysis and modulates glucose and lipid metabolism. It is known that the high-temperature (HT) treatment enhances corticosterone (CORT) secretion, muscle proteolysis, and mitochondrial reactive oxygen species (mtROS) generation in chickens. The present study investigated the coeffects of CORT on proteolysis and mtROS production, together with glucose, fatty acid, and amino acid metabolism in HT-treated cells. Myoblast cells were isolated from the major pectoralis muscle of five 0- or 1-day-old neonatal chicks and were precultured at 37 degrees C/CO2 conditions for 48 h to reach subconfluent (80%-90%) conditions. Cells were then reseeded onto a 6- or 24-well microplate for the subsequent measurement, followed by the culture under a control temperature (37 degrees C, control) or HT (41 degrees C) conditions for 1 or 6 h. The HT-treated cells were cocultured with physiologically relevant concentrations of CORT (20 ng/mL in dimethyl sulfoxide). The HT treatment decreased cellular protein content (P < 0.05) and increased atrogin-1 mRNA levels and mtROS generation levels compared to the control group (P < 0.05), whereas HT/CORT co-treatment did not induce changes in either parameter. The mRNA level of glucose transporter-1 was decreased in HT-treated cells compared to that in normal cells (P < 0.05), and the decrease was increased in the CORT co-treatment (P < 0.05). While HT treatment did not alter pyruvate dehydrogenase kinase-4 mRNA level, the level was increased in the CORT co-treatment compared to the control and HT-treated cells (P < 0.05). Neither HT nor HT/CORT treatments altered the mRNA levels of fatty acid oxidation-related factors, carnitine palmitoyl transferase-1, and cluster of differentiation 36. The study conducted a metabolic analysis using gas chromatography-mass spectrometry. The results showed that HT/CORT-treated cells had decreased intracellular citrate and alpha-ketoglutarate levels (P < 0.05) and increased extracellular alanine and amino acid that have gluconeogenic properties, as well as increased aspartate, isoleucine, serine, methionine, and threonine levels (P < 0.05) compared to HT-treated cells. These results suggest that CORT may not affect proteolysis and mtROS production but can suppress pyruvate oxidation and promote alanine production in HT-treated chickens. (C) 2021 Elsevier Inc. All rights reserved.