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首页> 外文期刊>Biology of Reproduction: Offical Journal of the Society for the Study of Reproduction >Functional Roles of Arginine During the Peri-Implantation Period of Pregnancy. III. Arginine Stimulates Proliferation and Interferon Tau Production by Ovine Trophectoderm Cells via Nitric Oxide and Polyamine-TSC2-MTOR Signaling Pathways
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Functional Roles of Arginine During the Peri-Implantation Period of Pregnancy. III. Arginine Stimulates Proliferation and Interferon Tau Production by Ovine Trophectoderm Cells via Nitric Oxide and Polyamine-TSC2-MTOR Signaling Pathways

机译：精氨酸在妊娠围产期的功能作用。三，精氨酸刺激一氧化氮和多胺-TSC2-MTOR信号通路通过绵羊滋养外胚层细胞增殖和干扰素Tau的生产。

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In mammal species, arginine is a multifunctional amino acid required for survival, growth, and development of conceptuses (embryo/fetus and associated extraembryonic membranes) during the peri-implantation period of pregnancy. However, functional roles of arginine with respect to it being a substrate for production of nitric oxide (NO) and polyamines on trophectoderm cell proliferation and function remain largely unknown. To systematically assess roles of arginine in conceptus development and its effect on interferon tau (IFNT) production for pregnancy recognition signaling in ruminants, an established ovine trophectoderm (oTr1) cell line isolated from Day-15 ovine conceptuses were used to determine their response to arginine, putrescine, and NO donors, as well as their associated inhibitors. Arginine at physiological concentration (0.2 mM) stimulated maximum oTr cell proliferation (increased 2.0-fold at 48 h and 2.6-fold at 96 h; P < 0.05), stimulated IFNT production (IFNT/cell increased 3.1-fold; P < 0.05), and increased total protein per cell by more than 1.5-fold (P < 0.05). It also increased phosphorylated tuberous sclerosis protein (p-TSC2) and phosphorylated mechanistic target of rapamycin (MTOR) abundance by more than 2.7- and 4.3-fold (P < 0.0001) after long-term incubation, respectively. When N omega-nitro-L-arginine methyl ester hydrochloride (L-NAME; NO synthase inhibitor), DL-alpha-difluoromethylornithine hydrochloride hydrate (DFMO; ornithine decarboxylase inhibitor), and the combination (L-NAME + DFMO) were added, the effects of arginine on cell proliferation was reduced by 10.7%, 16.1%, and 22.3% (P < 0.05) at 48 h, and 15.3%, 27.2%, and 39.1% (P < 0.05) at 96 h of incubation, respectively, but values remained 1.5-fold higher (P < 0.05) than for the arginine-free control, which suggests that arginine, per se, serves as a growth factor. Both putrescine and NO stimulate cell proliferation via activation of the TSC2-MTOR signaling cascade, whereas only putrescine increased IFNT production. Collectively, our results indicate that arginine is essential for oTr1 cell proliferation and IFNT production via the NO/polyamine-TSC2-MTOR signaling pathways, particularly the pathway involving polyamine biosynthesis.
机译：在哺乳动物物种中，精氨酸是在怀孕的围植入期生存，生长和繁殖概念（胚胎/胎儿和相关的胚外膜）所需的多功能氨基酸。然而，关于精氨酸作为滋养外胚层细胞增殖和功能的一氧化氮（NO）和多胺生产底物的功能作用仍然是未知的。为了系统评估精氨酸在概念发育中的作用及其对反刍动物中妊娠识别信号的干扰素tau（IFNT）产生的影响，使用从第15天绵羊概念中分离出的已建立的绵羊滋养外胚层（oTr1）细胞系来确定其对精氨酸的反应，腐胺和NO供体及其相关抑制剂。生理浓度（0.2 mM）的精氨酸刺激最大的oTr细胞增殖（在48小时增加2.0倍，在96小时增加2.6倍; P <0.05），刺激IFNτ产生（IFNT /细胞增加3.1倍; P <0.05） ，并使每个细胞的总蛋白质增加1.5倍以上（P <0.05）。长期孵育后，它还能使雷帕霉素的磷酸化结节性硬化蛋白（p-TSC2）和磷酸化的机械靶标（MTOR）丰度分别提高2.7倍和4.3倍以上（P <0.0001）。当添加Nω-硝基-L-精氨酸甲酯盐酸盐（L-NAME; NO合酶抑制剂），DL-α-二氟甲基鸟氨酸盐酸盐水合物（DFMO;鸟氨酸脱羧酶抑制剂）和组合（L-NAME + DFMO）时，在培养48小时后，精氨酸对细胞增殖的影响分别降低了10.7％，16.1％和22.3％（P <0.05），在培养96小时后分别降低了15.3％，27.2％和39.1％（P <0.05）。 ，但其值仍比不含精氨酸的对照高1.5倍（P <0.05），这表明精氨酸本身就是生长因子。腐胺和NO均通过激活TSC2-MTOR信号级联反应来刺激细胞增殖，而仅腐胺会增加IFNτ的产生。总体而言，我们的结果表明，精氨酸通过NO /多胺-TSC2-MTOR信号传导途径，特别是涉及多胺生物合成的途径，对oTr1细胞增殖和IFNτ产生至关重要。

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