Giuliani Alessandro, Lorenzini Luca, Gallamini Michele, Massella Alessandro, Giardino Luciana, Calzà Laura
BioPharmaNet-DIMORFIPA, University of Bologna, Via Tolara di Sopra 50, 40064 Ozzano dell'Emilia, Bologna, Italy.
BMC Complement Altern Med. 2009 Apr 15;9:8. doi: 10.1186/1472-6882-9-8.
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-à-vis two distinct biological effects: neurite elongation under NGF stimulus on laminin-collagen substrate and cell viability during oxidative stress.
We used a 670 nm laser, with extremely low peak power output (3 mW/cm2) and at an extremely low dose (0.45 mJ/cm2). 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.
We found that laser irradiation stimulates NGF-induced neurite elongation on a laminin-collagen coated substrate and protects PC12 cells against oxidative stress.
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.
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广泛关注。考虑到可见辐射在光谱中远红到近红外区域的临床应用,我们研究了具有极低能量传递的相干红光照射对源自大鼠嗜铬细胞瘤的神经细胞系的影响。我们重点关注脉冲光激光照射对两种不同生物学效应的影响:在层粘连蛋白 - 胶原蛋白底物上NGF刺激下的神经突伸长以及氧化应激期间的细胞活力。
我们使用了一台670纳米的激光,其峰值功率输出极低(3毫瓦/平方厘米)且剂量极低(0.45毫焦/平方厘米)。在培养的三天内测量神经突伸长。通过使用JC1活性染料和激光共聚焦显微镜实时记录线粒体膜电位(MMP),在不存在(光漂白)和存在(氧化应激)过氧化氢的情况下,以及通过MTT细胞活力测定,评估相干红光照射对细胞氧化应激反应的影响。
我们发现激光照射可刺激层粘连蛋白 - 胶原蛋白包被底物上NGF诱导的神经突伸长,并保护PC12细胞免受氧化应激。
这些数据表明,如MMP测量和MTT测定所示,红光辐射在氧化应激情况下可保护细胞培养物的活力。它还刺激神经突生长,并且这种效应可能对轴突保护也有积极意义。