Gajewski Michał, Gajewska Joanna, Rzodkiewicz Przemysław, Wojtecka-Łukasik Elżbieta
Department of Biochemistry and Molecular Biology, National Institute of Geriatrics, Rheumatology and Rehabilitation, Warsaw, Poland.
Screening Department, Institute of Mother and Child, Warsaw, Poland.
Reumatologia. 2016;54(3):103-7. doi: 10.5114/reum.2016.61209. Epub 2016 Jul 18.
Leptin is an adipose cells derived hormone that regulates energy homeostasis within the body. Energy metabolism of immune cells influences their activity within numerous pathological states, but the effect of leptin on these cells in unclear. On the one hand, it was observed that leptin induces neutrophils chemotaxis and modulates phagocytosis. On the other hand, neutrophils exposed to leptin did not display detectable Ca(2+) ions mobilization or β2-integrin upregulation. In this study, we investigated the effect of leptin on the redox homeostasis in lymphocytes and neutrophils.
Neutrophils and lymphocytes were isolated by density-gradient centrifugation of blood from healthy volunteers. Cells were cultured with or without leptin (100 ng/ml for lymphocytes and 500 ng/ml for neutrophils) or with or without synovial fluid (85%) for 0-72 h. Culture media were not changed during incubation. Cells were homogenized and homogenate was frozen until laboratory measurements. Redox homeostasis was assessed by the reduced glutathione (GSH) vs. oxidized glutathione (GSSG) ratio and membrane lipid peroxidation evaluation.
Lymphocytes cultured with leptin and synovial fluid showed a significant increase of the GSSG level. The GSSG/GSH ratio increased by 184 ±37%. In neutrophils incubated in a similar environment, the GSSG/GSH ratio increased by just 21 ±7%, and the effect was observed irrespectively of whether they were exposed to leptin or synovial fluid or both together. Neither leptin nor synovial fluid influenced lipid peroxidation in neutrophils, but in lymphocytes leptin intensified lipid peroxidation.
Leptin altered the lymphocytes, but not neutrophils redox state. Because firstly neutrophils are anaerobic cells and have just a few mitochondria and secondly lymphocytes have typical aerobic metabolism, the divergence of our data supports the hypothesis that leptin induces oxidative stress by modulation of mitochondria.
瘦素是一种由脂肪细胞分泌的激素，可调节体内能量平衡。免疫细胞的能量代谢会影响其在多种病理状态下的活性，但瘦素对这些细胞的影响尚不清楚。一方面，观察到瘦素可诱导中性粒细胞趋化并调节吞噬作用。另一方面，暴露于瘦素的中性粒细胞未表现出可检测到的钙离子动员或β2整合素上调。在本研究中，我们调查了瘦素对淋巴细胞和中性粒细胞氧化还原稳态的影响。
通过密度梯度离心法从健康志愿者血液中分离出中性粒细胞和淋巴细胞。细胞在有或无瘦素（淋巴细胞为100 ng/ml，中性粒细胞为500 ng/ml）或有或无滑液（85%）的条件下培养0 - 72小时。培养期间不更换培养基。将细胞匀浆，匀浆液冷冻保存直至进行实验室检测。通过还原型谷胱甘肽（GSH）与氧化型谷胱甘肽（GSSG）的比值以及膜脂质过氧化评估来评估氧化还原稳态。
用瘦素和滑液培养的淋巴细胞显示GSSG水平显著升高。GSSG/GSH比值增加了184±37%。在类似环境中培养的中性粒细胞中，GSSG/GSH比值仅增加了21±7%，且无论它们是暴露于瘦素、滑液还是两者同时存在，均观察到该效应。瘦素和滑液均未影响中性粒细胞的脂质过氧化，但在淋巴细胞中瘦素加剧了脂质过氧化。
瘦素改变了淋巴细胞而非中性粒细胞的氧化还原状态。由于一方面中性粒细胞是厌氧细胞且线粒体较少，另一方面淋巴细胞具有典型的有氧代谢，我们的数据差异支持了瘦素通过调节线粒体诱导氧化应激的假说。
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