亮氨酸对猪胎盘滋养层细胞增殖及氨基酸转运的影响

为研究亮氨酸(Leu)对猪胎盘滋养层细胞(pTr)增殖、凋亡以及氨基酸转运载体表达的影响及其机制,本试验用不同浓度Leu(0、1、10 mmol/L)分别处理pTr细胞24 h和48 h后,使用荧光定量PCR技术检测pTr细胞增殖和凋亡相关基因、氨基酸转运载体以及mTOR信号通路关键蛋白等的mRNA表达水平。结果表明:Leu处理pTr细胞24 h后,1 mmol/L试验组的SNAT1(P<0.01)、4E-BP1 (P<0.05)和eIF4G(P<0.05)的mRNA相对表达量低于对照组;Leu处理pTr细胞48 h后,1 mmol/L试验组LAT1(P<0.05)、4E-BP1(P<0.01)的mRNA相对表达量低于对照组,10 mmol/L试验组CDK4(P<0.05)、4E-BP1 (P <0.01)、SNAT1 (P <0.01)、SNAT2 (P <0.01)、LAT1 (P <0.01)以及rBAT (P <0.05)的mRNA相对表达量也低于对照组;Leu处理pTr细胞24 h和48 h后,10 mmol/L组mTORC1的mRNA相对表达量较对照组和1 mmol/L组均极显著提高(P<0.01)。可见,10 mmol/L Leu会抑制pTr细胞的增殖活力,并可能通过mTOR信号通路的介导,降低了pTr细胞氨基酸转运载体的表达。

Effect and Mechanism of Leucine on Proliferation and Amino Acid Transport in Porcine Trophectoderm Cell

To investigate the effects and mechanisms of leucine on proliferation, apoptosis and regulation of amino acid transporters in porcine trophectoderm cells(pTr), pTr cells were treated with different concentrations of Leu(0, 1, 10 mmol/L) for 24 h and 48 h in this experiment. The mRNA of pTr cell proliferation and apoptosis-related genes, amino acid transporter, the key proteins in mTOR signaling pathway were detected by real-time PCR. The results showed that the relative expression levels of SNAT1(P<0.01), 4 E-BP1(P<0.05) and eIF4 G(P<0.05) in the 1 mmol/L test group were lower than those in the control group after treatment with Leu for 24 h. After treatment of pTr cells for 48 h, the relative expression levels of LAT1(P<0.05) and 4 EBP1(P<0.01) in the 1 mmol/L test group were lower than those in the control group, and 10 mmol/L test group CDK4(P<0.05). 4 E-BP1(P<0.01), SNAT1(P<0.01), SNAT2(P<0.01), LAT1(P<0.01), and rBAT(P<0.05) mRNA expression levels were also lower than the control group; Leu treatment After 24 h and 48 h of pTr cells, the relative expression of mTORC1 mRNA in 10 mmol/L group was significantly higher than that in the control group and 1 mmol/L group(P<0.01). All the currently available data demonstrate that 10 mmol/L Leu can suppress the proliferation of pTr and reduces the expression of amino acid transporters in pTr cells by affecting the mTOR signaling pathway.