Lymphocyte proliferation modulated by glutamine: involved in the endogenous redox reaction
Lymphocyte proliferation modulated by glutamine: involved in the endogenous redox reaction
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DOI：
10.1046/j.1365-2249.1999.01009.x
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发表时间：
1999-09-01
影响因子：
4.6
通讯作者：
Shaio, MF
中科院分区：
文献类型：
--
作者：
Chang, WK;Yang, KD;Shaio, MF
谷氨酰胺浓度降低是在分解代谢应激过程中发现的，与感染易感性有关。然而，对谷氨酰胺调节淋巴细胞功能的机制知之甚少。谷氨酰胺不仅是线粒体中重要的能量来源，而且是谷氨酸的前体，其用于淋巴细胞中细胞谷胱甘肽（GSH）的生物合成。在这项研究中，我们研究了谷氨酰胺对淋巴细胞增殖过程中氧化还原反应的影响。从健康成年志愿者中获得的外周血单个核细胞在不同谷氨酰胺浓度的存在下培养并由植物血凝素（PHA）刺激。收获细胞并制备用于分析淋巴细胞增殖、细胞周期增殖、细胞内谷胱甘肽水平和活性氧（ROS）产生。我们发现，谷氨酰胺补充剂显著增强PHA刺激的淋巴细胞增殖和细胞周期从G（1）到S和G（2）/M期的传播。谷氨酰胺还增强PHA刺激的淋巴细胞中细胞内ROS和CSII水平的产生。通过汞橙子染色法进行的犁流式细胞术分析表明，谷氨酰胺显著增强PHA刺激的CD 4（+）淋巴细胞亚群中的细胞内非蛋白巯基，但不增强CD 8（+）淋巴细胞亚群中的细胞内非蛋白巯基。此外，用一氯二亚胺染料探针检测细胞内GSH，结果表明谷氨酰胺对PHA刺激的CD 4（+）和CD 8（+）淋巴细胞亚群的GSH均有增强作用。谷氨酰胺补充不足导致淋巴细胞增殖减少，细胞内GSH水平降低。添加外源性GSH显着增强淋巴细胞增殖，而GSH合成的阻断增强活性氧的产生和抑制淋巴细胞增殖。这些结果表明谷氨酰胺对PHA刺激的淋巴细胞增殖的调节与维持适当的细胞内氧化还原状态密切相关。
Decreased glutamine concentrations are found during catabolic stress and are related to susceptibility to infections. However, little is known about the mechanism of glutamine modulation of lymphocyte functions. Glutamine is not only an important energy source in mitochondria, but is also a precursor of glutamate, which is used for cellular glutathione (GSH) biosynthesis in lymphocytes. In this study, we investigated the effects of glutamine on the redox reaction during lymphocyte proliferation. Peripheral blood mononuclear cells, obtained from healthy adult volunteers, were cultured and stimulated by phytohaemagglutinin (PHA) in the presence of different glutamine concentrations. Cells were harvested and prepared for analysis of lymphocyte proliferation, cell cycle propagation, intracellular glutathione levels and reactive oxygen species (ROS) production. We found that glutamine supplementation significantly enhanced PHA-stimulated lymphocyte proliferation and propagation of the cell cycle from the G(1) to S and G(2)/M phases. Glutamine also enhanced production of both intracellular ROS and CSII levels in PHA-stimulated lymphocytes. Plow cytometric analysis by the mercury orange staining method showed that glutamine significantly enhanced intracellular non-protein thiols in PHA-stimulated CD4(+), but not CD8(+) lymphocyte subsets. Furthermore, intracellular GSH detected by monochlorobimane dye probe showed that glutamine enhanced GSH both in PHA-stimulated CD4(+) and CD8(+) lymphocyte subsets. Inadequate glutamine supplementation resulted in decreased lymphocyte proliferation in association with decreased levels of intracellular GSH. Addition of exogenous GSH significantly enhanced lymphocyte proliferation, whereas blockade of GSH synthesis enhanced ROS production and suppressed lymphocyte proliferation. These results suggest that the modulation of PHA-stimulated lymphocyte proliferation by glutamine is closely related to the maintenance of appropriate intracellular redox status.