PMA 처리한 인간 불멸화 상피세포에서 청색 및 적색 광 조사에 따른 활성산소 제거에 대한 비교 연구 A Comparative Study of Blue and Red Light Irradiation on PMA-Treated HaCaT Cell Line on ROS Scavenging EWffect
- 언어KOR
- URLhttp://db.koreascholar.com/Article/Detail/292905
pp.27-36
Intracellular reactive oxygen species(ROS) produced in a various pathologic state was known to intermediate many cellular response such as inflammation. Recently, low level light irradiation by HeNe laser used in many clinical field could improve inflammatory state by scavenging intracellular ROS through photo-detachment/dissociation process. The purpose of this study is to investigate the differential effects of blue and red light irradiation on ROS scavenging effects. Immortalized human oral keratinocyte HaCat cells were used. Phorbol 12-myristate 13-acetate(PMA) was treated for inflammation. Red(635nm) and blue(470nm) light irradiation was carried out. To asses the intracellular ROS by light irradiation, confocal microscopic and flow cytometric assay with DCF fluorescence for total ROS and ESR spectrometry of DMPO-O2 - for superoxide anion were caried out. And microarray was performed for mRNA expression level. Released intracellular total ROS in PMA treated HaCat cell lines was dissociated efficiently by red light irradiation, while blue light irradiation did not. Rather, blue light irradiation increased ROS formation. For superoxide anion generated the first synthetic form of ROS, red light irradiation reduced its amount but blue light irradiation did not. In the mRNA expression in line with cyclooxygenase(COX) pathway, prostagrandin endoperoxide synthase 1(PTGS 1), prostagrandin endoperoxide synthase 2(PTGS 2) and phospholipase A2(PLA2) were increased by both light irradiation and they were decreased as time flows. And genes associated with ROS releasing, mRNA expressions of tumor necrosis factor receptor (TNFR) and interleukin 1beta(IL1B) were increased by 1 hour red light irradiation but did not by blue light irradiation. As a result, red and blue light irradiation showed different response in affecting the level of ROS. These findings indicate that red light rather than blue light is more useful for anti-inflammation in clinical field
II. MATERIALS AND METHODS
1. Primary cell culture and chemicals
2. Light source and irradiation
3. Enzyme-linked immunoassay for PGE2
4. Flow cytometer and laser scanning confocalmicroscope analysis for detectionof ROS formation
5. Electron spin resonance(ESR) spin trappingdetermination of superoxide anion(O2-)
6. Total RNA isolation
7. Analysis of mRNA expression by microarraychip
8. Statistical Data Analysis
III. RESULTS
1. PGE2 release and ROS detection
2. Microarray analysis
IV. DISCUSSION
V. CONCLUSIONS
VI. REFERENCES