PDF(Pubmed)
Abstract：
Regulation of mechanistic target of rapamycin complex 1 (mTORC1) plays an important role in aging and nutrition. For example, caloric restriction reduces mTORC1 signaling and extends lifespan, whereas nutrient abundance and obesity increase mTORC1 signaling and reduce lifespan. Skeletal muscle-specific knockout (KO) of DEP domain-containing 5 protein (DEPDC5) results in constitutively active mTORC1 signaling, muscle hypertrophy and an increase in mitochondrial respiratory capacity. The metabolic profile of skeletal muscle, in the setting of hyperactive mTORC1 signaling, is not well known.
To determine the metabolomic and lipidomic signature in skeletal muscle from female and male wild-type (WT) and DEPDC5 KO mice.
Tibialis anterior (TA) muscles from WT and transgenic (conditional skeletal muscle-specific DEPDC5 KO) were obtained from female and male adult mice. Polar metabolites and lipids were extracted using a Bligh-Dyer extraction from 5 samples per group and identified and quantified by LC-MS/MS. Resulting analyte peak areas were analyzed with t-test, analysis of variance, and Volcano plots for group comparisons (e.g., WT compared with KO) and multivariate statistical analysis for genotype and sex comparisons.
A total of 162 polar metabolites (organic acids, amino acids, and amines and acyl carnitines) and 1141 lipid metabolites were detected in TA samples by LC-MS/MS. Few polar metabolites showed significant differences in KO muscles compared with WT within the same sex group. P-aminobenzoic acid, β-alanine, and dopamine were significantly higher in KO male muscle whereas erythrose-4-phosphate and oxoglutaric acid were significantly reduced in KO females. The lipidomic profile of the KO groups revealed an increase of muscle phospholipids and reduced triacylglycerol and diacylglycerol compared with the WT groups.
Sex differences were detected in polar metabolome and lipids were dependent on genotype. The metabolomic profile of mice with hyperactive skeletal muscle mTORC1 is consistent with an upregulation of mitochondrial function and amino acid utilization for protein synthesis.
To determine the metabolomic and lipidomic signature in skeletal muscle from female and male wild-type (WT) and DEPDC5 KO mice.
Tibialis anterior (TA) muscles from WT and transgenic (conditional skeletal muscle-specific DEPDC5 KO) were obtained from female and male adult mice. Polar metabolites and lipids were extracted using a Bligh-Dyer extraction from 5 samples per group and identified and quantified by LC-MS/MS. Resulting analyte peak areas were analyzed with t-test, analysis of variance, and Volcano plots for group comparisons (e.g., WT compared with KO) and multivariate statistical analysis for genotype and sex comparisons.
A total of 162 polar metabolites (organic acids, amino acids, and amines and acyl carnitines) and 1141 lipid metabolites were detected in TA samples by LC-MS/MS. Few polar metabolites showed significant differences in KO muscles compared with WT within the same sex group. P-aminobenzoic acid, β-alanine, and dopamine were significantly higher in KO male muscle whereas erythrose-4-phosphate and oxoglutaric acid were significantly reduced in KO females. The lipidomic profile of the KO groups revealed an increase of muscle phospholipids and reduced triacylglycerol and diacylglycerol compared with the WT groups.
Sex differences were detected in polar metabolome and lipids were dependent on genotype. The metabolomic profile of mice with hyperactive skeletal muscle mTORC1 is consistent with an upregulation of mitochondrial function and amino acid utilization for protein synthesis.
摘要：
背景：mTORC1的调节在衰老和营养中起重要作用。例如,热量限制减少mTORC1信号并延长寿命，而营养丰富和肥胖会增加mTORC1信号传导并缩短寿命。骨骼肌特异性敲除含有DEP结构域的5蛋白（DEPDC5）导致组成型活性mTORC1信号，肌肉肥大和线粒体呼吸能力增加。骨骼肌的代谢概况，在mTORC1信号传导过度活跃的情况下，不是众所周知的。
目的：确定雌性和雄性野生型和DEPDC5基因敲除小鼠骨骼肌的代谢组学和脂质组学特征。
方法：来自野生型（WT）和转基因（条件性骨骼肌特异性DEPDC5敲除，KO）从雌性（F）和雄性（M）成年小鼠获得。使用Bligh-Dyer从每组5个样品中提取极性代谢物和脂质，并通过LC-MS/MS进行鉴定和定量。所得分析物峰面积用t检验分析,方差分析,和用于组比较的火山图(例如，WTvsKO)和基因型和性别比较的多变量统计分析。
结果：总共162种极性代谢物（有机酸，通过LC-MS/MS检测TA样品中的氨基酸和胺以及酰基肉碱）和1141脂质代谢物。与同性别组的WT相比，很少有极性代谢物在KO肌肉中显示出显着差异。对氨基苯甲酸,KO男性肌肉中的β-丙氨酸和多巴胺显着升高，而KO女性中的4-磷酸红细胞和氧戊二酸显着降低。与WT组相比，KO组的脂质组学特征显示肌肉磷脂增加，三酰甘油和二酰甘油减少。
结论：在极性代谢组中检测到性别差异，血脂依赖于基因型。骨骼肌mTORC1过度活跃的小鼠的代谢组学特征与线粒体功能和氨基酸对蛋白质合成的利用的上调一致。
目的：确定雌性和雄性野生型和DEPDC5基因敲除小鼠骨骼肌的代谢组学和脂质组学特征。
方法：来自野生型（WT）和转基因（条件性骨骼肌特异性DEPDC5敲除，KO）从雌性（F）和雄性（M）成年小鼠获得。使用Bligh-Dyer从每组5个样品中提取极性代谢物和脂质，并通过LC-MS/MS进行鉴定和定量。所得分析物峰面积用t检验分析,方差分析,和用于组比较的火山图(例如，WTvsKO)和基因型和性别比较的多变量统计分析。
结果：总共162种极性代谢物（有机酸，通过LC-MS/MS检测TA样品中的氨基酸和胺以及酰基肉碱）和1141脂质代谢物。与同性别组的WT相比，很少有极性代谢物在KO肌肉中显示出显着差异。对氨基苯甲酸,KO男性肌肉中的β-丙氨酸和多巴胺显着升高，而KO女性中的4-磷酸红细胞和氧戊二酸显着降低。与WT组相比，KO组的脂质组学特征显示肌肉磷脂增加，三酰甘油和二酰甘油减少。
结论：在极性代谢组中检测到性别差异，血脂依赖于基因型。骨骼肌mTORC1过度活跃的小鼠的代谢组学特征与线粒体功能和氨基酸对蛋白质合成的利用的上调一致。
