Kamalnayan Bajaj Diabetology Research Centre, King Edward Memorial Hospital Research Centre, Pune, India.
Department of Molecular Medicine, Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Cleveland, OH, United States of America.
PLoS One. 2018 May 24;13(5):e0196970. doi: 10.1371/journal.pone.0196970. eCollection 2018.
Methionine loading test (MLT) has been used primarily to identify defects in transsulfuration of homocysteine in cystathionine beta synthase deficiency. It may not be as useful to evaluate remethylation pathway, in vitamin B-12 and folate deficiencies.
We used tracer isotope labelled MLT to interrogate transsulfuration and remethylation independently in vitamin B-12 deficiency.
We studied vitamin B-12 deficient women with a tracer labelled MLT before and eleven months after treatment with vitamin B-12. The fractional contribution of [13C]homocysteine to breath CO2 was used as a measure of transsulfuration, and difference in the intracellular enrichment of [13C]methionine and that of [C2H3]methionine as a measure of remethylation of homocysteine. Combined pre- and post-treatment results were analyzed to investigate the association between plasma vitamin B-12 concentrations and measures of homocysteine metabolism.
The subjects were 17 years old, with a BMI of 19.4 kg/m2. Treatment with vitamin B-12, 2µg/day increased plasma B-12 from 93 (78.7, 106.2) [median (25th, 75th centiles)] to 161.5 (125.5, 226.2) pmol/L; 44% were below <150pmol/L after treatment. Fasting homocysteine concentration was significantly lower and that of cysteine higher in subjects with B-12 levels >150pmol/L. The tracer estimated transsulfuration of homocysteine was lower and remethylation higher with B-12 levels >150pmol/L when compared with those <150pmol/L.
The tracer labelled MLT in combination with fasting parameters is a robust way to estimate parameters of methionine metabolism and can be used in the field where prime-constant rate infusion studies cannot be done efficiently.
蛋氨酸负荷试验(MLT)主要用于鉴定胱硫醚 β 合酶缺乏症中同型半胱氨酸转硫过程的缺陷。它在评估维生素 B12 和叶酸缺乏症中的再甲基化途径时可能不太有用。
我们使用示踪同位素标记的 MLT 来独立研究维生素 B12 缺乏症中的转硫作用和再甲基化作用。
我们研究了维生素 B12 缺乏的女性,在接受维生素 B12 治疗前和治疗 11 个月后进行了示踪标记的 MLT。[13C]同型半胱氨酸呼气 CO2 的分数贡献被用作转硫作用的衡量标准,[13C]蛋氨酸和 [C2H3]蛋氨酸的细胞内富集差异被用作衡量同型半胱氨酸再甲基化的指标。分析治疗前后的综合结果,以研究血浆维生素 B12 浓度与同型半胱氨酸代谢指标之间的关系。
研究对象年龄为 17 岁,BMI 为 19.4kg/m2。每天 2µg 的维生素 B12 治疗使血浆 B12 从 93(78.7,106.2)[中位数(25 分位,75 分位)]增加到 161.5(125.5,226.2)pmol/L;治疗后 44%的患者血浆 B12 水平仍<150pmol/L。B12 水平>150pmol/L 的受试者空腹同型半胱氨酸浓度明显较低,半胱氨酸浓度较高。与 B12 水平<150pmol/L 的患者相比,示踪剂估计的同型半胱氨酸转硫作用较低,再甲基化作用较高。
结合空腹参数的示踪标记 MLT 是一种可靠的方法,可以用来估计蛋氨酸代谢的参数,并且可以在无法有效进行基础恒速输注研究的情况下使用。