Greco M, Vacca R A, Moro L, Perlino E, Petragallo V A, Marra E, Passarella S
Centro di Studio sui Mitocondri e Metabolismo Energetico, C.N.R., Via Amendola 165/A I-70126 BARI, Italy.
Lasers Surg Med. 2001;29(5):433-41. doi: 10.1002/lsm.1137.
To gain some insight into the photostimulation of isolated hepatocytes irradiated with Helium-Neon (He-Ne) laser light certain biochemical events were studied with respect to two mechanisms: i) the direct light dependent activation of certain biochemical events investigated in intact cells and isolated mitochondria, ii) the indirect stimulation of processes per se light independent.
STUDY DESIGNS/MATERIALS AND METHODS: Irradiation of either isolated hepatocytes or isolated rat liver mitochondria was carried out with He-Ne laser (wavelength, 632.8 nm; fluence, 0.24 J cm-2; fluence rate, 12 mW cm-2). Changes in mitochondrial membrane potential in isolated hepatocytes were monitored using the cationic probe safranine. The c-fos expression was studied by Northern blot and immunoblot analysis.
As a result of irradiation, increase of the mitochondrial membrane potential was found to occur in irradiated hepatocytes both in the presence or in the absence of CaCl2. The hyperpolarization of the mitochondrial membrane is assumed to cause an increase in mitochondrial Ca2+ uptake that was measured in isolated mitochondria. Finally, an increase in c-fos expression was found in irradiated hepatocytes when incubated in the presence of CaCl2.
This paper gives additional information on the mechanism by which He-Ne laser light, either directly or in a cascade-like effect dependent on increase in cell Ca2+, can cause cell stimulation.
为深入了解氦氖(He-Ne)激光照射对分离的肝细胞的光刺激作用,针对两种机制研究了某些生化事件:i)在完整细胞和分离的线粒体中研究的某些生化事件的直接光依赖性激活,ii)对本身与光无关的过程的间接刺激。
研究设计/材料与方法:用He-Ne激光(波长632.8nm;通量0.24J/cm²;通量率12mW/cm²)照射分离的肝细胞或分离的大鼠肝线粒体。使用阳离子探针番红监测分离的肝细胞中线粒体膜电位的变化。通过Northern印迹和免疫印迹分析研究c-fos表达。
照射结果显示,无论有无CaCl₂,照射的肝细胞中线粒体膜电位均升高。线粒体膜的超极化被认为会导致分离的线粒体中测量到的线粒体Ca²⁺摄取增加。最后,当在CaCl₂存在下孵育时,照射的肝细胞中c-fos表达增加。
本文提供了关于He-Ne激光直接或通过依赖于细胞Ca²⁺增加的级联效应导致细胞刺激的机制的更多信息。