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

作　　者：苗婧 李丰阳 王婷婷[2] 张美娜 李娜 黄晶[2] 雷连成 MIAO Jing;LI Fengyang;WANG Tingting;ZHANG Meina;LI Na;HUANG Jing;LEILiancheng(College of Veterinary Medicine,Jilin University,Changchun 130062,China;The First Bethune Hospital of Jilin University,Changchun 130061,China)

机构地区：[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

出　　处：《吉林农业大学学报》2024年第6期1049-1054,共6页Journal of Jilin Agricultural University

基　　金：国家自然科学基金面上项目(31772715)。

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。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 detect 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 knockdown of CARS2,the contents of arginine and citrulline in MAC-T cells were significantly downregulated,and the content of agmatine was up-regulated;Meantime,ASS1 was decreased,OTC expression 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 

分 类 号：S823[农业科学—畜牧学]