School of Biology and Basic Medical Sciences, Medical College, Soochow University, Suzhou 215123, China.
Institute of Agricultural Biotechnology &Ecology (IABE), Soochow University, Suzhou 215123, China.
Sci Rep. 2017 Apr 10;7:46258. doi: 10.1038/srep46258.
Abnormalities in the circadian clock system are known to affect the body's metabolic functions, though the molecular mechanisms responsible remain uncertain. In this study, we achieved continuous knockdown of B. mori Period (BmPer) gene expression in the B. mori ovary cell line (BmN), and generated a Per-KD B. mori model with developmental disorders including small individual cells and slow growth. We conducted cell metabolomics assays by gas chromatography/liquid chromatography-mass spectrometry and showed that knockdown of BmPer gene expression resulted in significant inhibition of glycometabolism. Amino acids that used glucose metabolites as a source were also down-regulated, while lipid metabolism and nucleotide metabolism were significantly up-regulated. Metabolite correlation analysis showed that pyruvate and lactate were closely related to glycometabolism, as well as to metabolites such as aspartate, alanine, and xanthine in other pathways. Further validation experiments showed that the activities of the key enzymes of glucose metabolism, hexokinase, phosphofructokinase, and citrate synthase, were significantly decreased and transcription of their encoding genes, as well as that of pyruvate kinase, were also significantly down-regulated. We concluded that inhibition of the circadian clock gene BmPer repressed glycometabolism, and may be associated with changes in cellular amino acid metabolism, and in cell growth and development.
昼夜节律钟系统的异常已知会影响身体的代谢功能,但负责的分子机制仍不确定。在这项研究中,我们在 B. mori 卵巢细胞系(BmN)中实现了对 B. mori Period(BmPer)基因表达的连续敲低,并生成了具有发育障碍的 Per-KD B. mori 模型,包括个体细胞小和生长缓慢。我们通过气相色谱/液相色谱-质谱法进行了细胞代谢组学分析,结果表明 BmPer 基因表达的敲低导致糖代谢的显著抑制。作为葡萄糖代谢物来源的氨基酸也下调,而脂质代谢和核苷酸代谢则显著上调。代谢物相关性分析表明,丙酮酸和乳酸与糖代谢密切相关,与其他途径中的天冬氨酸、丙氨酸和黄嘌呤等代谢物也密切相关。进一步的验证实验表明,葡萄糖代谢的关键酶,如己糖激酶、磷酸果糖激酶和柠檬酸合酶的活性显著降低,其编码基因以及丙酮酸激酶的转录也显著下调。我们得出结论,昼夜节律钟基因 BmPer 的抑制抑制了糖代谢,可能与细胞氨基酸代谢以及细胞生长和发育的变化有关。