13C-NMR study of hypoglycemia-induced glycolytic changes in embryonic mouse heart.

13C-NMR study of hypoglycemia-induced glycolytic changes in embryonic mouse heart.
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低血糖引起的胚胎小鼠心脏糖酵解变化的 13C-NMR 研究。

DOI:
10.1002/tera.10103
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发表时间:
2002
期刊:
Teratology.
影响因子:
--
通讯作者:
Smoak,IdaW
Smoak,IdaW
中科院分区:
--
文献类型:
--
作者:
Ghatnekar,GautamS;Gracz,HannaS;Smoak,IdaW

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背景利用13 C-NMR光谱可以检测胚胎小鼠组织中的葡萄糖代谢产物。这种方法的优点是在其化学特异性和跟踪代谢变化的能力。MethodsIn this study,CD-1小鼠交配和胚胎切除妊娠日(GD)10.5(plug = GD 0.5)。分离心脏并在150 mg/dl葡萄糖(正常血糖培养基)或40 mg/dl葡萄糖(低血糖培养基)中培养6 h。13 C标记的葡萄糖占培养基中总葡萄糖的62%-64%。培养前和培养后的培养基用氘代水(D2 O)处理,13 C光谱使用Bruker Avance 500 MHz光谱仪在11.744特斯拉(125.7 MHz用于13 C)下获得。NMR光谱显示13 C-葡萄糖在预培养正常血糖和低血糖培养基中的共振。正常血糖和低血糖培养基的培养后光谱显示了13 C-葡萄糖信号以及13 C-乳酸信号。测量来自预培养培养基的[1 - 13 C-葡萄糖]共振和来自培养后培养基的[3 - 13 C-乳酸]共振的曲线下面积(AUC)。计算培养后[3 - 13 C-乳酸]与培养前[1 - 13 C-葡萄糖]的AUC比值,发现低血糖组高于血糖正常组。ResultsOur结果证实了早期使用放射性标记底物的发现,并表明13 C-NMR光谱可用于研究暴露于低血糖的离体胚胎心脏的葡萄糖代谢。在相同的光谱中,从而确定暴露于低血糖和正常血糖后离体胚胎心脏中的代谢通量。该方法可用于评价其他致畸性暴露后胚胎组织的葡萄糖代谢。畸形学66:267-272,2002年。© 2002 Wiley利斯公司
BackgroundGlucose metabolites can be detected in embryonic mouse tissues using13C‐NMR spectroscopy. The advantage of this method is in its chemical specificity and the ability to follow metabolic changes.MethodsIn this study, CD‐1 mice were mated and embryos excised on gestational day (GD) 10.5 (plug = GD 0.5). Hearts were isolated and cultured in 150 mg/dl glucose (normoglycemic medium) or 40 mg/dl glucose (hypoglycemic medium) for 6 hr.13C‐labeled glucose comprised 62%–64% of total glucose in the culture medium. Pre‐ and postculture media were treated with deuterated water (D2O), and13C spectra were obtained using a Bruker Avance 500 MHz spectrometer operating at 11.744 tesla (125.7 MHz for13C). NMR spectra demonstrated resonances for13C‐glucose in preculture normoglycemic and hypoglycemic media. Postculture spectra for normoglycemic and hypoglycemic media demonstrated13C‐glucose signals as well as a signal for13C‐lactate. Area under the curve (AUC) was measured for the [1‐13C‐glucose] resonance from preculture media and the [3‐13C‐lactate] resonance from postculture media. The ratios of AUC for postculture [3‐13C‐lactate] to preculture [1‐13C‐glucose] were calculated and found to be higher in hypoglycemic than in normoglycemic media.ResultsOur results confirm earlier findings using radiolabeled substrates and suggest that13C‐NMR spectroscopy can be used to study glucose metabolism in isolated embryonic hearts exposed to hypoglycemia.ConclusionsNMR effectively measures glucose and its metabolite, lactate, in the same spectrum and thus determines metabolic flux in the isolated embryonic heart after exposure to hypoglycemia and normoglycemia. This method could evaluate glucose metabolism in embryonic tissues following other teratogenic exposures. Teratology 66:267–272, 2002. © 2002 Wiley‐Liss, Inc.
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DOI: --
发表时间: 1981
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影响因子: --
作者:
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DOI: --
发表时间: 1983
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Leung,LL;Harpel,PC;Nachman,RL;Rabellino,EM
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DOI: 10.1515/bchm2.1972.353.2.1125
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DOI: --
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