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首页> 外文期刊>Journal of dairy science >Short communication: The effect of increasing concentrations of different methionine forms and 2-hydroxy-4-(methylthio)butanoic acid on genes controlling methionine metabolism in primary bovine neonatal hepatocytes
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Short communication: The effect of increasing concentrations of different methionine forms and 2-hydroxy-4-(methylthio)butanoic acid on genes controlling methionine metabolism in primary bovine neonatal hepatocytes

机译：短期通信：将不同甲硫氨酸形式和2-羟基-4-（甲基硫基-4-（甲基）丁酸在原发性牛新生儿肝细胞中甲硫氨酸代谢基因上增加的影响

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The d-isomer of Met cannot be used directly by themammary gland in dairy cows; instead, it is transformedinto l-Met, the proteogenic isomer, in the liverand other extramammary tissues. It remains unclearwhether different Met forms and a Met hydroxy analog,2-hydroxy-4-(methylthio)butanoic acid (HMB), are metabolizedand function similarly in the liver. The objectiveof the present study was to examine the regulationof key genes in Met regeneration, transulfuration, andtransmethylation pathways in response to increasingdoses of different Met forms. Hepatocytes isolated from4 calves between 4 and 7 d old were maintained asmonolayer cultures for 24 h before addition of treatments.Treatments of (0, 10, 20, 40 μM) d-Met, l-Met,dl-Met, dl-HMB, or a 1:1 mixture of dl-Met anddl-HMB were added to Met-free medium in triplicate.After 24 h, cell lysates were collected for quantificationof gene expression by quantitative PCR, and mRNAabundance was normalized to the mean of 3 referencegenes. Data were analyzed with PROC MIXED of SAS9.3 (SAS Institute Inc., Cary, NC). Analyses of covarianceconfirmed equivalent slopes of Met form, and thefinal model included form, dose, and random effect ofcalf within form. Data are reported as least squaresmeans ± standard error. No main effect of Met formwas observed for any genes examined. The enzymesencoded by betaine-homocysteine methyltransferase(BHMT) and 5-methyltetrahydrofolate-homocysteinemethyltransferase use betaine and 5-methyltetrahydrofolate,respectively, to regenerate Met from homocysteine.Increasing concentration of Met did not alter 5-methyltetrahydrofolateexpression, but decreased BHMTexpression. Expression of glycine N-methyltransferase,the enzyme that controls transmethylation flux fromS-adenosyl-methionine, was not affected by Met concentration.Methionine concentration had no effect onexpression of cystathionine β-synthase, a key enzymefor the transulfuration pathway. The decrease in BHMTexpression indicates a decreased need for cellular Metregeneration with increasing Met concentration, independentof Met form. The lack of differences amongMet forms on regulating genes examined indicates thatall Met forms similarly reduced genes controlling Metregeneration and metabolism in primary bovine hepatocytes.
机译：遇见的D-异构体不能直接使用乳制奶牛的乳腺;相反，它被转变为进入L-MET，肝脏异构体，在肝脏中和其他肾上腺组织。它仍然不清楚无论是不同的MET表格和满足的羟基类似物，2-羟基-4-（甲硫基）丁酸（HMB）是代谢的并且在肝脏中同样起作用。目标是本研究是检查规定符合再生，过束率和均衡的关键基因响应越来越多的酰基化途径不同符合不同的形式的剂量。肝细胞被隔绝4和7 d旧的4个犊牛保持为在加入治疗之前24小时的单层培养物。（0,10,20,40μm）D-Met，L-Met的处理，DL-MET，DL-HMB或1：1的DL-MET混合物和将DL-HMB一式三份加入无培养基中。24小时后，收集细胞裂解物以定量通过定量PCR和mRNA的基因表达丰度被标准化为3个参考的平均值基因。通过SAS混合分析数据进行分析9.3（SAS Institute Inc.，Cary，NC）。协方差分析确认了符合符合形式的等效斜率，以及最终模型包括形式，剂量和随机效果表格中的小牛。数据报告为最小二乘法表示±标准错误。没有符合符合的主要效果观察到检查的任何基因。酶由甜菜碱 - 同型半胱氨酸甲基转移酶编码（BHMT）和5-甲基四氢氟醇 - 同型半胱氨酸甲基转移酶使用甜菜碱和5-甲基四氢氟醇，分别从同型半胱氨酸再生达到。持续浓度的持浓度没有改变5-甲基四氢氟醇表达，但减少了bhmt表达。甘氨酸N-甲基转移酶的表达，从而控制液体乙基化通量的酶S-腺苷 - 甲硫氨酸不受符合浓度的影响。甲硫氨酸浓度没有影响胱硫胺β-合酶的表达，键酶用于过度熔断途径。 BHMT的减少表达表明蜂窝符合的需求减少再生随着普及浓度，独立达到的形式。缺乏差异在检查的调节基因上满足表明表明所有符合表明同样减少了控制的基因原发性牛肝细胞的再生和代谢。

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