Low infra red laser light irradiation on cultured neural cells: effects on mitochondria and cell viability after oxidative stress
Low infra red laser light irradiation on cultured neural cells: effects on mitochondria and cell viability after oxidative stress
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DOI：
10.1186/1472-6882-9-8
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发表时间：
2009-04-15
影响因子：
--
通讯作者：
Calza, Laura
中科院分区：
文献类型：
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作者：
Giuliani, Alessandro;Lorenzini, Luca;Calza, Laura
背景：近年来，生物系统对可见光敏感这一众所周知的概念引起了人们的极大兴趣。考虑到可见光辐射在远红外到近红外光谱区域的临床应用，我们探索了具有极低能量转移的相干红光照射对大鼠嗜铬细胞瘤神经细胞系的影响。我们专注于脉冲光激光照射的影响与两个不同的生物学效应：神经生长因子刺激下的神经突起延长层粘连蛋白胶原底物和细胞活力在氧化stress.Methods：我们使用670 nm的激光，具有极低的峰值功率输出（3 mW/cm（2））和极低的剂量（0.45 mJ/cm（2））。在培养物中在三天内测量神经突伸长。通过使用JC 1活体染料和激光共聚焦显微镜实时记录线粒体膜电位（MMP），在没有细胞膜电位的情况下，评价相干红光照射对细胞氧化应激反应的影响。（光漂白）和存在H2O2（氧化应激），并通过MTT细胞活力测定。我们发现激光照射刺激神经生长因子诱导的神经突起在层粘连蛋白-胶原包被的基底上的伸长，并保护PC12细胞免受氧化应激。这些数据表明，红光辐射在氧化应激的情况下保护细胞培养物的活力，如MMP测量和MTT测定所示。它还刺激神经突生长，这种作用也可能对轴突保护产生积极影响。
Background: Considerable interest has been aroused in recent years by the well-known notion that biological systems are sensitive to visible light. With clinical applications of visible radiation in the far-red to near-infrared region of the spectrum in mind, we explored the effect of coherent red light irradiation with extremely low energy transfer on a neural cell line derived from rat pheochromocytoma. We focused on the effect of pulsed light laser irradiation vis-a-vis two distinct biological effects: neurite elongation under NGF stimulus on laminin-collagen substrate and cell viability during oxidative stress.Methods: We used a 670 nm laser, with extremely low peak power output (3 mW/cm(2)) and at an extremely low dose (0.45 mJ/cm(2)). Neurite elongation was measured over three days in culture. The effect of coherent red light irradiation on cell reaction to oxidative stress was evaluated through live-recording of mitochondria membrane potential (MMP) using JC1 vital dye and laser-confocal microscopy, in the absence (photo bleaching) and in the presence (oxidative stress) of H2O2, and by means of the MTT cell viability assay.Results: We found that laser irradiation stimulates NGF-induced neurite elongation on a laminin-collagen coated substrate and protects PC12 cells against oxidative stress.Conclusion: These data suggest that red light radiation protects the viability of cell culture in case of oxidative stress, as indicated by MMP measurement and MTT assay. It also stimulates neurite outgrowth, and this effect could also have positive implications for axonal protection.