Effects of Cysteinyl-tRNA Synthetase 2 on Arginine Metabolism and Autophagy in Bovine Mammary Epithelial Cells
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.
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