低强度氦氖（He-Ne）激光对α轻型地中海贫血患者红细胞影响的光镊拉曼光谱研究
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摘要
目的:利用激光光镊拉曼光谱系统,研究人血正常红细胞与α轻型地中海贫血患者血红细胞之间的差异,测定低强度氦氖(He-Ne)激光对单个红细胞照射前后的拉曼光谱变化,探讨了低强度He-Ne激光对正常红细胞和α轻型地贫患者红细胞的影响作用及机理。
方法:取健康献血者、α轻型地贫患者新鲜血液标本,利用MR450型低强度氦氖(He-Ne)激光仪,采取不同功率(2mW～5mW)和不同照射时间(40s、10min、30min、1h)的激光对红细胞悬液进行照射。用激光光镊拉曼光谱系统获得激光照射红细胞前后的拉曼光谱。
结果:1.与正常红细胞相比,α轻型地贫患者红细胞的整体谱线偏弱;部分的特征谱数发生位移;谱数为1212cm-1、1224cm-1融合为一个特征峰值1216cm-1。表明:α轻型地贫患者红细胞与正常红细胞光谱有一定区别。2.低强度He-Ne激光对正常红细胞的拉曼光谱影响作用不明显。3.低强度He-Ne激光对α轻型地贫患者红细胞影响:⑴照射组和对照组间比较:①照射组整体强度也有所减弱;②谱数1622cm-1和谱数1604 cm-1的相对强度增加,主要由于谱数1604cm-1强度减弱引起;③谱数1004cm-1强度变弱峰形变尖,半高宽减少。⑵不同照射时间具有不同的效应:①激光照射时间40s后1h,谱数754cm-1强度降到最低;②谱数1622cm-1与1604cm-1的相对强度在激光照射10min后1h发生明显变化,而在照射时间30min后30min时出现该效应,在照射1h后即刻出现这一效应。表明:照射时间越长,激光对红细胞苯丙氨酸平行振动模和酪氨酸拉伸模的影响效应出现越早,并且在静置1h后效应增加;③激光照射时间30min后1h,出现一个新峰值1226cm-1。
结论: 1.激光光镊拉曼光谱为探讨人血正常红细胞与α轻型地贫患者血红细胞之间的差异提供有用的信息;2.适当功率和适当照射时间的低强度He-Ne激光对α轻型地贫患者红细胞携氧功能有一定的调节作用,对细胞结构改变、键与键之间的扭转等都有一定的影响;这些影响在激光照射停止后仍维持一段时间,并可能达到最大效应;这表明了低强度激光对α轻型地贫患者红细胞作用具有较为明显的延迟效应和累积效应。
Objective: To make use of laser tweezers Raman spectroscopy system, Study the differences of red blood cells between normal human and patients withα-light thalassemia, and to discuss the effects and mechanism of low-intensity He-Ne laser on red blood cells from normal andα-thalassemia patients .
Methods: To take the fresh blood samples from healthy donors andα- light thalassemia patients,using MR450-type low-intensity He-Ne laser vertically irradiated red blood cell suspension by different power (2mW～5mW) and different irradiated time (40s, 10min, 30min, 1h) . To obtain Raman spectra of red blood cells by laser tweezers Raman spectroscopy system.
Results: 1.Comparred with normal RBC (red blood cells), overall intensity of Raman spectroscopy of RBC inα-light thalassemia is weak; Some characteristic peaks have happened a few displacement: spectrum of 1212 cm-1, 1224cm-1 integrated as one a feature of the peak 1216cm-1. Shows that there must some spectral difference of RBC betweenα-ight thalassemia and normal. 2. The effect of low-intensity He-Ne laser on normal RBC was not obvious in Raman spectra. 3. The effect of low-intensity He-Ne laser on RBC fromα-light thalassemia patients:⑴Comparred irradiated group with control group, the overall intensity of irradiated group is also weak; the relative intensity of 1622cm-1 and 1604cm-1 increased mainly because of the spectral 1604cm-1 intensity reduced; for spectral 1004cm-1,the intensity decreased and the peak shape sharped and FWHM reduced. (2) Different irradiated time has a different effect: an hour later after laser irradiation time 40 seconds,the intensity of 754cm-1 reached minimum; an hour later after irradiated 10min ,the relative intensity of the spectrum 1622cm-1 and 1604cm-1 significant reduced; and same effect has seen at 30min later after irradiation 30min and immediately after irradiation at 1h.
Conclusion: 1.Laser tweezers Raman spectroscopy provides useful information in the difference of red blood cells between normal human and patients withα-light thalassemia. 2.The appropriate power and the appropriate irradiated time of low-intensity laser play important part in the oxygen-carrying function of RBC fromα-light thalassemia patients, and also impacted cell structure. These effects after the cessation of laser irradiation for a certain period is still maintained, and may reached maxmoium, this indicates that the delayed effect and the cumulative effect of low-intensity laser on theα-light thalassemia red blood cells is more obvious.
方法:取健康献血者、α轻型地贫患者新鲜血液标本,利用MR450型低强度氦氖(He-Ne)激光仪,采取不同功率(2mW～5mW)和不同照射时间(40s、10min、30min、1h)的激光对红细胞悬液进行照射。用激光光镊拉曼光谱系统获得激光照射红细胞前后的拉曼光谱。
结果:1.与正常红细胞相比,α轻型地贫患者红细胞的整体谱线偏弱;部分的特征谱数发生位移;谱数为1212cm-1、1224cm-1融合为一个特征峰值1216cm-1。表明:α轻型地贫患者红细胞与正常红细胞光谱有一定区别。2.低强度He-Ne激光对正常红细胞的拉曼光谱影响作用不明显。3.低强度He-Ne激光对α轻型地贫患者红细胞影响:⑴照射组和对照组间比较:①照射组整体强度也有所减弱;②谱数1622cm-1和谱数1604 cm-1的相对强度增加,主要由于谱数1604cm-1强度减弱引起;③谱数1004cm-1强度变弱峰形变尖,半高宽减少。⑵不同照射时间具有不同的效应:①激光照射时间40s后1h,谱数754cm-1强度降到最低;②谱数1622cm-1与1604cm-1的相对强度在激光照射10min后1h发生明显变化,而在照射时间30min后30min时出现该效应,在照射1h后即刻出现这一效应。表明:照射时间越长,激光对红细胞苯丙氨酸平行振动模和酪氨酸拉伸模的影响效应出现越早,并且在静置1h后效应增加;③激光照射时间30min后1h,出现一个新峰值1226cm-1。
结论: 1.激光光镊拉曼光谱为探讨人血正常红细胞与α轻型地贫患者血红细胞之间的差异提供有用的信息;2.适当功率和适当照射时间的低强度He-Ne激光对α轻型地贫患者红细胞携氧功能有一定的调节作用,对细胞结构改变、键与键之间的扭转等都有一定的影响;这些影响在激光照射停止后仍维持一段时间,并可能达到最大效应;这表明了低强度激光对α轻型地贫患者红细胞作用具有较为明显的延迟效应和累积效应。
Objective: To make use of laser tweezers Raman spectroscopy system, Study the differences of red blood cells between normal human and patients withα-light thalassemia, and to discuss the effects and mechanism of low-intensity He-Ne laser on red blood cells from normal andα-thalassemia patients .
Methods: To take the fresh blood samples from healthy donors andα- light thalassemia patients,using MR450-type low-intensity He-Ne laser vertically irradiated red blood cell suspension by different power (2mW～5mW) and different irradiated time (40s, 10min, 30min, 1h) . To obtain Raman spectra of red blood cells by laser tweezers Raman spectroscopy system.
Results: 1.Comparred with normal RBC (red blood cells), overall intensity of Raman spectroscopy of RBC inα-light thalassemia is weak; Some characteristic peaks have happened a few displacement: spectrum of 1212 cm-1, 1224cm-1 integrated as one a feature of the peak 1216cm-1. Shows that there must some spectral difference of RBC betweenα-ight thalassemia and normal. 2. The effect of low-intensity He-Ne laser on normal RBC was not obvious in Raman spectra. 3. The effect of low-intensity He-Ne laser on RBC fromα-light thalassemia patients:⑴Comparred irradiated group with control group, the overall intensity of irradiated group is also weak; the relative intensity of 1622cm-1 and 1604cm-1 increased mainly because of the spectral 1604cm-1 intensity reduced; for spectral 1004cm-1,the intensity decreased and the peak shape sharped and FWHM reduced. (2) Different irradiated time has a different effect: an hour later after laser irradiation time 40 seconds,the intensity of 754cm-1 reached minimum; an hour later after irradiated 10min ,the relative intensity of the spectrum 1622cm-1 and 1604cm-1 significant reduced; and same effect has seen at 30min later after irradiation 30min and immediately after irradiation at 1h.
Conclusion: 1.Laser tweezers Raman spectroscopy provides useful information in the difference of red blood cells between normal human and patients withα-light thalassemia. 2.The appropriate power and the appropriate irradiated time of low-intensity laser play important part in the oxygen-carrying function of RBC fromα-light thalassemia patients, and also impacted cell structure. These effects after the cessation of laser irradiation for a certain period is still maintained, and may reached maxmoium, this indicates that the delayed effect and the cumulative effect of low-intensity laser on theα-light thalassemia red blood cells is more obvious.
引文
[1] Cenevive Chottard,Irina Kazanskaya,Mireille Bruschi.Eur J Biochem,2000 267:1050
[2] Alexei V Feofanov,Alexei I Grichine,Larissa A Shitova et a1.Biophys.J,2000,78:499
[3] Cheng Ji-Xin,Kevin Jia Y,Zheng Gengfeng et a1.Biophys.J.,2002,83:502
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[7] Yan Xun-ling,Wang Qiu-guo,Dong Rui-xin(闫循领,王秋国,董瑞新).Spectro6copy and Spectral Analysis(光谱学与光谱分析),(待发表)
[8] PuppelsG J,GarritsenH S,Segers-Nolten G M et a1.Biophys.J.,1991,60:1046
[9] Narma M Sijtserna , Arian G J Tibbe , Segers-Nolten G M J et a1.Biophys.J.,2000,78:2606
[10]余国滔.拉曼光谱在医学上的应用[J].光散射学报,1996,8(3):174～180
[11] G. J. Puppels, H. S. P. Garritsen, G. M. J. Segers-Nolten, et a1.Raman microspectroscopic approach to the study of human granulocytes[J]. Biophys. J. c Biophysical Society,1992,60:1046-1056.
[12]岳粮跃,姚辉璐,莫华,等.光镊捕获的单个解冻人血红细胞的拉曼光谱研究[J].生物物理学,2007,23(1):75-79
[13]岳粮跃,莫华,姚辉璐,等.人血红细胞在直流电作用后光镊操纵的拉曼光谱研究[J].生物医学工程学杂志,2007,24(2):404-408
[14] Xie C A,Dinno M A.Li Y Q.Near-infrared Raman spectroscopy of single optically trapped biological cells[J].Opt Lett,2002,27:249.
[15] Xie C A,Li Y Q.Confocal micro-Raman spectroscopy of single biological cell using optical trapping and shifted excitation diference tehniques[J].Appl Phys,2003,93:2982.
[16] Xie C A,Li Y Q,Tang W.et a1.Study of dynamical process of heat denaturation in opticallytraIIped single micro organisms by near-infrared Raman spectroscopy[J].Appl Phys,2003,94:6138.
[17] Iijima K,Shimoyarna N,Shimoyarrm M,et a1.Effect of low power He—Ne laser on deformability of stored human erythrocytes[J].JClin LaserMed Surg,1993,1l(4):185-189
[18] Yova D.Antagonistic effects of epinephrine and helium—neon (He—Ne)laser irradiation on red blood cells deformability[J].Clin Hemorheol,1994,14(3):369-378.
[19]降雨强,王晓波,冯玫,等.He—Ne激光照射对兔血液携氧能力影响的实验研究[J]. 2004,21(5):270-271
[20]陈英华,池景全,陈敏,等.低强度氦氖激光调整红细胞变形性的机理研.应用激光,2001,21(4):271-274
[21]闫循领,董瑞新,王秋国.人血单个红细胞的共振拉曼光谱研究[J].光谱学与光谱分析,2004,24(5):576-578
[22]宓现强,岑剡,周正谊,等.低强度激光照射对离体动物红细胞流变学性质的影响[J].中国激光,2004,31(7):889-891
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[2]丁艳,郭海,顾文军.红细胞免疫在肿瘤中的进展[J].辽宁中医药大学学报,2007 ,9(1):68-69.
[3]Nelson RA.The immune adherence phenomenon[J].Science,1953,118(3077):733-737.
[4]Siegel I,LiuliuT,Gleicher N.The red-cell immune system[J].Lancet,1981,1(8246):556-558.
[5]Marilyn J.Erythrocyte adhesion receptors: blood group antigens and related molecules [J].Transfus Med Rev,2005,19(1):32-44.
[6] Nielsen CH,Matthiesen SH,Lyng I,et al.The role of complement receptor type 1 (CR1,CD35) in determining the cellular distribution of opsonized immune complexes between whole blood cells: kinetic analysis of the buffering capacity of erythrocytes [J].Immunology,1997,90 (1):129-137.
[7] Inada Y, Kamiyama M, Kanemitsu T, et al.In vivo binding of circulating immune complexes by C3b receptors (CR1) of transfused erythrocytes[J].Annals of the Rheumatic Diseases,1989,48(4):287-294.
[8]王彩虹.红细胞免疫调控研究进展[J].中国兽医杂志,2002,38(2):28-31
[9]郭峰.红细胞免疫及调节功能测定方法[J].免疫学杂志,1990,6(1):60-65.
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[2] Alexei V Feofanov,Alexei I Grichine,Larissa A Shitova et a1.Biophys.J,2000,78:499
[3] Cheng Ji-Xin,Kevin Jia Y,Zheng Gengfeng et a1.Biophys.J.,2002,83:502
[4] Zhao Hongxia,Xu Yi-ming,Zhang Zhi-yi.Chinese Science Bulletin(科学通报),1998,43(9):955
[5] Xu Yi-ming,Zhang Zhi-yi,Zhang Wei.Science in ChinaC(中国科学C),1998,28(5):417
[6] Sun Chuan-Wen,Xu Yi-zhuang,Sun Kai-hua et al(孙传文,徐怡庄,孙开华等).Spectroscopy and Spectral Analysis(光谱学与光谱分析),1996,16(5):22
[7] Yan Xun-ling,Wang Qiu-guo,Dong Rui-xin(闫循领,王秋国,董瑞新).Spectro6copy and Spectral Analysis(光谱学与光谱分析),(待发表)
[8] PuppelsG J,GarritsenH S,Segers-Nolten G M et a1.Biophys.J.,1991,60:1046
[9] Narma M Sijtserna , Arian G J Tibbe , Segers-Nolten G M J et a1.Biophys.J.,2000,78:2606
[10]余国滔.拉曼光谱在医学上的应用[J].光散射学报,1996,8(3):174～180
[11] G. J. Puppels, H. S. P. Garritsen, G. M. J. Segers-Nolten, et a1.Raman microspectroscopic approach to the study of human granulocytes[J]. Biophys. J. c Biophysical Society,1992,60:1046-1056.
[12]岳粮跃,姚辉璐,莫华,等.光镊捕获的单个解冻人血红细胞的拉曼光谱研究[J].生物物理学,2007,23(1):75-79
[13]岳粮跃,莫华,姚辉璐,等.人血红细胞在直流电作用后光镊操纵的拉曼光谱研究[J].生物医学工程学杂志,2007,24(2):404-408
[14] Xie C A,Dinno M A.Li Y Q.Near-infrared Raman spectroscopy of single optically trapped biological cells[J].Opt Lett,2002,27:249.
[15] Xie C A,Li Y Q.Confocal micro-Raman spectroscopy of single biological cell using optical trapping and shifted excitation diference tehniques[J].Appl Phys,2003,93:2982.
[16] Xie C A,Li Y Q,Tang W.et a1.Study of dynamical process of heat denaturation in opticallytraIIped single micro organisms by near-infrared Raman spectroscopy[J].Appl Phys,2003,94:6138.
[17] Iijima K,Shimoyarna N,Shimoyarrm M,et a1.Effect of low power He—Ne laser on deformability of stored human erythrocytes[J].JClin LaserMed Surg,1993,1l(4):185-189
[18] Yova D.Antagonistic effects of epinephrine and helium—neon (He—Ne)laser irradiation on red blood cells deformability[J].Clin Hemorheol,1994,14(3):369-378.
[19]降雨强,王晓波,冯玫,等.He—Ne激光照射对兔血液携氧能力影响的实验研究[J]. 2004,21(5):270-271
[20]陈英华,池景全,陈敏,等.低强度氦氖激光调整红细胞变形性的机理研.应用激光,2001,21(4):271-274
[21]闫循领,董瑞新,王秋国.人血单个红细胞的共振拉曼光谱研究[J].光谱学与光谱分析,2004,24(5):576-578
[22]宓现强,岑剡,周正谊,等.低强度激光照射对离体动物红细胞流变学性质的影响[J].中国激光,2004,31(7):889-891
[1]Denomme GA. The structure and function of the molecules that carry human red blood cell and platelet antigens[J]. Transfus Med Rev,2004,18(3):203-231.
[2]丁艳,郭海,顾文军.红细胞免疫在肿瘤中的进展[J].辽宁中医药大学学报,2007 ,9(1):68-69.
[3]Nelson RA.The immune adherence phenomenon[J].Science,1953,118(3077):733-737.
[4]Siegel I,LiuliuT,Gleicher N.The red-cell immune system[J].Lancet,1981,1(8246):556-558.
[5]Marilyn J.Erythrocyte adhesion receptors: blood group antigens and related molecules [J].Transfus Med Rev,2005,19(1):32-44.
[6] Nielsen CH,Matthiesen SH,Lyng I,et al.The role of complement receptor type 1 (CR1,CD35) in determining the cellular distribution of opsonized immune complexes between whole blood cells: kinetic analysis of the buffering capacity of erythrocytes [J].Immunology,1997,90 (1):129-137.
[7] Inada Y, Kamiyama M, Kanemitsu T, et al.In vivo binding of circulating immune complexes by C3b receptors (CR1) of transfused erythrocytes[J].Annals of the Rheumatic Diseases,1989,48(4):287-294.
[8]王彩虹.红细胞免疫调控研究进展[J].中国兽医杂志,2002,38(2):28-31
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