Sharma Sulbha K, Kharkwal Gitika B, Sajo Mari, Huang Ying-Ying, De Taboada Luis, McCarthy Thomas, Hamblin Michael R
Wellman Center for Photomedicine, Massachusetts General Hospital, Department of Dermatology, Harvard Medical School, Boston, Massachusetts 02114, USA.
Lasers Surg Med. 2011 Sep;43(8):851-9. doi: 10.1002/lsm.21100.
In the past four decades numerous studies have reported the efficacy of low level light (laser) therapy (LLLT) as a treatment for diverse diseases and injuries. Recent studies have shown that LLLT can biomodulate processes in the central nervous system and has been extensively studied as a stroke treatment. However there is still a lack of knowledge on the effects of LLLT at the cellular level in neurons. The present study aimed to study the effect of 810 nm laser on several cellular processes in primary cortical neurons cultured from embryonic mouse brains.
STUDY DESIGN/MATERIALS AND METHODS: Neurons were irradiated with fluences of 0.03, 0.3, 3, 10, or 30 J/cm(2) of 810-nm laser delivered over varying times at 25 mW/cm(2) and intracellular levels of reactive oxygen species (ROS), nitric oxide and calcium were measured using fluorescent probes within 5 minutes of the end of irradiation. The changes in mitochondrial function in response to light were studied in terms of adenosine triphosphate (ATP) and mitochondrial membrane potential (MMP).
Light induced a significant increase in calcium, ATP and MMP at lower fluences and a decrease at higher fluences. ROS was significantly induced at low fluences, followed by a decrease and a second larger increase at 30 J/cm(2). Nitric oxide levels showed a similar pattern of a double peak but values were less significant compared to ROS.
The results suggest that LLLT at lower fluences is capable of inducing mediators of cell signaling processes which in turn may be responsible for the beneficial stimulatory effects of the low level laser. At higher fluences beneficial mediators are reduced and high levels of Janus-type mediators such as ROS and NO (beneficial at low concentrations and harmful at high concentrations) may be responsible for the damaging effects of high-fluence light and the overall biphasic dose response.
在过去的四十年里,众多研究报道了低强度光(激光)疗法(LLLT)对多种疾病和损伤的治疗效果。近期研究表明,LLLT可对中枢神经系统的过程进行生物调节,并且作为一种中风治疗方法已得到广泛研究。然而,关于LLLT在神经元细胞水平上的作用仍缺乏了解。本研究旨在探讨810纳米激光对从小鼠胚胎脑培养的原代皮质神经元中几种细胞过程的影响。
研究设计/材料与方法:用25毫瓦/平方厘米的功率,在不同时间内给予神经元0.03、0.3、3、10或30焦/平方厘米的810纳米激光照射,在照射结束后5分钟内使用荧光探针测量细胞内活性氧(ROS)、一氧化氮和钙的水平。从三磷酸腺苷(ATP)和线粒体膜电位(MMP)方面研究光对线粒体功能的影响。
低能量密度时,光诱导钙、ATP和MMP显著增加,高能量密度时则降低。低能量密度时ROS显著诱导产生,随后下降,在30焦/平方厘米时再次大幅增加。一氧化氮水平呈现类似的双峰模式,但与ROS相比,其值的变化不太显著。
结果表明,低能量密度的LLLT能够诱导细胞信号转导过程的介质,这反过来可能是低强度激光产生有益刺激作用的原因。在高能量密度时,有益介质减少,而高水平的Janus型介质如ROS和NO(低浓度时有益,高浓度时有害)可能是高能量密度光产生损伤作用以及整体双相剂量反应的原因。