半胱氨酰tRNA合成酶2对奶牛乳腺上皮细胞精氨酸代谢及细胞自噬的影响
Effects of Cysteinyl-tRNA Synthetase 2 on Arginine Metabolism and Autophagy in Bovine Mammary Epithelial Cells
苗婧 1李丰阳 1王婷婷 2张美娜 1李娜 1黄晶 2雷连成1
作者信息
- 1. 吉林大学动物医学学院,长春 130062
- 2. 吉林大学白求恩第一医院,长春 130061
- 折叠
摘要
为探讨半胱氨酰tRNA合成酶2(Cysteinyl-tRNA synthetase 2,CARS2)对奶牛乳腺上皮细胞精氨酸代谢及细胞自噬的影响,采用RNA干扰敲降CARS2,高效液相色谱质谱联用(HPLC-MS)检测奶牛乳腺上皮细胞系MAC-T细胞内精氨酸等相关氨基酸代谢变化;采用ELISA及Western blot检测CARS2对MAC-T细胞精氨酸代琥珀酸合成酶1(Argininosuccinate synthetase 1,ASS1)、鸟氨酸氨甲酰基转移酶(Ornithinetranscarbamylase,OTC)、精氨酸脱羧酶(Arginine decarboxylase,ADC)的表达或酶活力变化;用qPCR及Western blot检测CARS2对MAC-T细胞自噬的影响.结果表明:RNAi敲降CARS2后,MAC-T细胞内精氨酸、瓜氨酸的含量显著下调,胍丁胺的含量上调;ASS1降低,OTC表达量及酶活力均降低,ADC表达量及酶活力升高;同时发现CARS2可以调控MAC-T细胞自噬的发生.说明CARS2下调可以加速MAC-T细胞精氨酸代谢并促进细胞自噬的产生,CARS2可能是人工干预精氨酸代谢及其相关疾病的潜在靶点.
Abstract
To explore the effects of Cysteinyl-tRNA synthetase 2(CARS2)on arginine metabolism and autophagy in bovine mammary epithelial cells,RNA interference was used to knock down CARS2,and high performance liquid chromatography-mass spectrometry(HPLC-MS)was used to de-tect the metabolic changes of arginine and other related amino acids in bovine mammary epithelial cell line MAC-T cells;ELISA and Western blot were used to determine the effects of CARS2 on the expression or enzyme activity of argininosuccinate synthase 1(ASS1),ornithine carbamoyltransferase(OTC)and arginine decarboxylase(ADC)in MAC-T cells.The effect of CARS2 on autophagy in MAC-T cells was detected by qPCR and Western blotting.The results showed that after RNAi knock-down of CARS2,the contents of arginine and citrulline in MAC-T cells were significantly down-regulated,and the content of agmatine was up-regulated;Meantime,ASS1 was decreased,OTC expres-sion and enzymatic activity were also decreased,while ADC expression and enzymatic activity were increased.CARS2 was also found to regulate the occurrence of autophagy in MAC-T cell,indicating that down-regulation of CARS2 can accelerate arginine metabolism in MAC-T cells and promote the appearance of autophagy and CARS2 may be a potential target for artificially intervening arginine metabolism to improve milk quality.
关键词
CARS2/精氨酸代谢/细胞自噬/奶牛乳腺上皮细胞Key words
CARS2/arginine metabolism/autophagy/bovine mammary epithelial cell引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据