Basal fatty acid oxidation increases after recurrent low glucose in human primary astrocytes.
Basal fatty acid oxidation increases after recurrent low glucose in human primary astrocytes.
复制标题
DOI：
10.1007/s00125-018-4744-6
复制
发表时间：
2019-01
期刊：
影响因子：
8.2
通讯作者：
Beall C
中科院分区：
文献类型：
--
作者：
Weightman Potter PG;Vlachaki Walker JM;Robb JL;Chilton JK;Williamson R;Randall AD;Ellacott KLJ;Beall C
低血糖是1型糖尿病患者血糖控制良好的主要障碍。频繁的低血糖发作会损害对后续低血糖发作的意识。支持低血糖意识的神经变化定义不清，神经胶质细胞参与低血糖感知和葡萄糖反调节的分子机制需要进一步研究。本研究的目的是检测急性和复发性低血糖（RLG）是否以及通过何种机制改变了人类原发性星形胶质细胞（HPA）的功能。为了测试神经胶质细胞，特别是星形胶质细胞，是否可以检测到葡萄糖的变化，我们利用HPA和U373星形细胞瘤细胞，并将它们暴露于体外RLG。这允许以高特异性和灵敏度测量RLG相关的细胞代谢变化。我们使用蛋白质印迹法检查蛋白质磷酸化/表达的变化。使用Seahorse细胞外通量分析仪评估代谢功能。免疫荧光成像被用来检查细胞形态和酶测定被用来测量乳酸释放，糖原含量，细胞内ATP和核苷酸的比例。AMP活化蛋白激酶（AMPK）在病理生理相关的葡萄糖浓度范围内被激活。RLG产生的基础线粒体代谢的脂肪酸氧化的依赖性增加，并表现出线粒体应激的标志，包括质子泄漏增加和耦合效率降低。相对于葡萄糖的可用性，乳酸盐的释放增加，在低血糖，但这是没有修改RLG。基础葡萄糖摄取没有修改RLG和糖原水平相似，在控制和RLG处理的细胞。RLG暴露后，通过维持血糖正常水平，线粒体对RLG的适应部分恢复。综上所述，这些数据表明，HPA线粒体改变后RLG，与代谢开关增加脂肪酸氧化，这表明神经胶质细胞适应RLG涉及改变线粒体代谢，可能有助于缺陷的葡萄糖反调节糖尿病低血糖。本文的在线版本（10.1007/s 00125 -018-4744-6）包含同行评审但未经编辑的补充材料，可供授权用户使用。
Hypoglycaemia is a major barrier to good glucose control in type 1 diabetes. Frequent hypoglycaemic episodes impair awareness of subsequent hypoglycaemic bouts. Neural changes underpinning awareness of hypoglycaemia are poorly defined and molecular mechanisms by which glial cells contribute to hypoglycaemia sensing and glucose counterregulation require further investigation. The aim of the current study was to examine whether, and by what mechanism, human primary astrocyte (HPA) function was altered by acute and recurrent low glucose (RLG). To test whether glia, specifically astrocytes, could detect changes in glucose, we utilised HPA and U373 astrocytoma cells and exposed them to RLG in vitro. This allowed measurement, with high specificity and sensitivity, of RLG-associated changes in cellular metabolism. We examined changes in protein phosphorylation/expression using western blotting. Metabolic function was assessed using a Seahorse extracellular flux analyser. Immunofluorescent imaging was used to examine cell morphology and enzymatic assays were used to measure lactate release, glycogen content, intracellular ATP and nucleotide ratios. AMP-activated protein kinase (AMPK) was activated over a pathophysiologically relevant glucose concentration range. RLG produced an increased dependency on fatty acid oxidation for basal mitochondrial metabolism and exhibited hallmarks of mitochondrial stress, including increased proton leak and reduced coupling efficiency. Relative to glucose availability, lactate release increased during low glucose but this was not modified by RLG. Basal glucose uptake was not modified by RLG and glycogen levels were similar in control and RLG-treated cells. Mitochondrial adaptations to RLG were partially recovered by maintaining euglycaemic levels of glucose following RLG exposure. Taken together, these data indicate that HPA mitochondria are altered following RLG, with a metabolic switch towards increased fatty acid oxidation, suggesting glial adaptations to RLG involve altered mitochondrial metabolism that could contribute to defective glucose counterregulation to hypoglycaemia in diabetes. The online version of this article (10.1007/s00125-018-4744-6) contains peer-reviewed but unedited supplementary material, which is available to authorised users.
登录
查看更多内容
影响因子：
8.1
作者：
Buckman LB;Thompson MM;Lippert RN;Blackwell TS;Yull FE;Ellacott KL
通讯作者：
Ellacott KL
影响因子：
64.5
作者：
García-Cáceres C;Quarta C;Varela L;Gao Y;Gruber T;Legutko B;Jastroch M;Johansson P;Ninkovic J;Yi CX;Le Thuc O;Szigeti-Buck K;Cai W;Meyer CW;Pfluger PT;Fernandez AM;Luquet S;Woods SC;Torres-Alemán I;Kahn CR;Götz M;Horvath TL;Tschöp MH
通讯作者：
Tschöp MH
影响因子：
5.3
作者：
Levy, LM;Warr, O;Attwell, D
通讯作者：
Attwell, D
影响因子：
16.2
作者：
Brazeau, Anne-Sophie;Rabasa-Lhoret, Remi;Strychar, Irene;Mircescu, Hortensia
通讯作者：
Mircescu, Hortensia
来自小鼠大脑多个区域的老化星形胶质细胞转录组。
影响因子：
8.8
作者：
Boisvert MM;Erikson GA;Shokhirev MN;Allen NJ
通讯作者：
Allen NJ