论文部分内容阅读
目的观察蛋氨酸(methionine,Met)负载对大鼠肝细胞同型半胱氨酸代谢酶及相关化合物的影响。方法体外培养BRL(Baffalo rat liver)细胞,将其分为两组,对照组和Met组(20 mmol/L Met作用1h)。采用高效液相色谱法检测同型半胱氨酸(Hcy)、半胱氨酸(Cys)、谷胱甘肽(GSH)、S-腺苷蛋氨酸(SAM)、S-腺苷同型半胱氨酸(SAH)含量,全自动氨基酸分析仪检测培养液中丝氨酸(Ser)、谷氨酸(Glu)、甘氨酸(Gly)、胱氨酸(Cys)、牛磺酸(Tau)的含量,参照文献方法检测细胞内S-腺苷同型半胱氨酸水解酶(SAHH)、胱硫醚-β-合成酶(CBS)、甜菜碱同型半胱氨酸甲基转移酶(BHMT)和蛋氨酸合成酶(MS)的活性。结果蛋氨酸负载后,培养液中Hcy、Cys、Ser和胱氨酸含量显著增加(P<0.05),而Glu、Gly和Tau含量显著减少(P<0.05);细胞内SAM含量显著增加(P<0.05),SAHH活性显著增强(P<0.05),BHMT和MS活性显著减弱(P<0.05)。SAH含量和CBS活性变化不明显。结论蛋氨酸负载可活化大鼠肝细胞中同型半胱氨酸的转硫化途径,同时抑制其再甲基化途径。
Objective To investigate the effects of methionine (Met) on the homocysteine metabolism enzymes and related compounds in rat hepatocytes. Methods BRL (Baffalo rat liver) cells were cultured in vitro and divided into two groups. The control group and the Met group (20 mmol / L Met for 1 h). Hcy, Cys, GSH, SAM, S-adenosylhomocysteine (Hcy) were detected by HPLC. (SAH), and the contents of Ser, Glu, Cys and Tau in the culture medium were detected by automatic amino acid analyzer. Reference literature The levels of intracellular S-adenosylhomocysteine hydrolase (SAHH), cystathionine-β-synthase (CBS), betaine homocysteine methyltransferase (BHMT) and methionine synthase ) Activity. Results The contents of Hcy, Cys, Ser and cystine in the culture medium were significantly increased (P <0.05), but Glu, Gly and Tau were significantly decreased (P <0.05) 0.05), SAHH activity was significantly increased (P <0.05), BHMT and MS activity was significantly decreased (P <0.05). SAH content and CBS activity did not change significantly. Conclusion Methionine load can activate the homocysteine transsulfuration pathway in rat hepatocytes and inhibit its remethylation pathway.