Oliveira P F, Alves M G, Rato L, Laurentino S, Silva J, Sá R, Barros A, Sousa M, Carvalho R A, Cavaco J E, Socorro S
CICS-UBI, Health Sciences Research Centre, University of Beira Interior, 6201-506 Covilhã, Portugal.
Biochim Biophys Acta. 2012 Feb;1820(2):84-9. doi: 10.1016/j.bbagen.2011.11.006. Epub 2011 Nov 20.
Sertoli cells metabolize glucose producing lactate for developing germ cells. As insulin regulates glucose uptake and its disturbance/insensitivity is associated with diabetes mellitus, we aimed to determine the effect of insulin deprivation in human Sertoli cell (hSC) metabolism and metabolism-associated gene expression.
hSC-enriched primary cultures were maintained in the absence/presence of insulin and metabolite variations were determined by (1)H-NMR. mRNA expression levels of glucose transporters (GLUT1, GLUT3), lactate dehydrogenase (LDHA) and monocarboxylate transporter (MCT4) were determined by RT-PCR.
Insulin deprivation resulted in decreased lactate production and in decrease of glucose consumption that was completely reverted after 6h. Cells of both groups consumed similar amounts of glucose. In insulin-deprived cells, transcript levels of genes associated to lactate metabolism (LDHA and MCT4) were decreased. Transcript levels of genes involved in glucose uptake exhibited a divergent variation: GLUT3 levels were decreased while GLUT1 levels increased. Insulin-deprived hSCs presented: 1) altered glucose consumption and lactate secretion; 2) altered expression of metabolism-associated genes involved in lactate production and export; 3) an adaptation of glucose uptake by modulating the expression of GLUT1 and GLUT3.
This is the first report regarding the effect of insulin-deprivation on hSC metabolism.
支持细胞代谢葡萄糖为发育中的生殖细胞产生乳酸。由于胰岛素调节葡萄糖摄取，且其紊乱/不敏感与糖尿病相关，我们旨在确定胰岛素缺乏对人支持细胞（hSC）代谢及代谢相关基因表达的影响。
在有无胰岛素的情况下维持富含hSC的原代培养物，并通过氢核磁共振（1H-NMR）测定代谢物变化。通过逆转录聚合酶链反应（RT-PCR）测定葡萄糖转运蛋白（GLUT1、GLUT3）、乳酸脱氢酶（LDHA）和单羧酸转运蛋白（MCT4）的mRNA表达水平。
胰岛素缺乏导致乳酸生成减少和葡萄糖消耗减少，6小时后完全恢复。两组细胞消耗的葡萄糖量相似。在胰岛素缺乏的细胞中，与乳酸代谢相关的基因（LDHA和MCT4）的转录水平降低。参与葡萄糖摄取的基因转录水平呈现不同变化：GLUT3水平降低而GLUT1水平升高。胰岛素缺乏的hSC表现为：1）葡萄糖消耗和乳酸分泌改变；2）参与乳酸生成和输出的代谢相关基因表达改变；3）通过调节GLUT1和GLUT3的表达来适应葡萄糖摄取。
这是关于胰岛素缺乏对hSC代谢影响的首篇报道。
