氢离子对血红蛋白-氧亲和力影响的拉曼光谱研究  

Raman Spectroscopy Study of the Effect of H^+ on the Oxygen Affinity Capacity of Hemoglobin

作　　者：刘培培[1] 关晓月[1] 曾常春[1,2] 聂广[3] 刘颂豪[4] 

机构地区：[1]华南师范大学生物光子学研究院,激光生命科学教育部重点实验室,广东广州510631 [2]桂林医学院基础医学院,广西桂林541004 [3]深圳市第三人民医院,广东深圳518112 [4]华南师范大学信息光电子科技学院,广东广州510631

出　　处：《光谱学与光谱分析》2013年第5期1234-1238,共5页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金项目(30873238);广东省科技计划项目(2007B031401010,2011B031700056)资助

摘　　要：静脉采血,分离并制备血红蛋白溶液,用拉曼光谱方法观察不同pH时H+对血红蛋白-氧结合状态与能力的影响。结果显示:在514.5nm激光的激发下,当pH为7.4时,随着氧分压降低,血红蛋白在1 375,1 562,1 585和1 638cm-1峰强均逐渐减弱,其中以1 375和1 638cm-1峰渐变最明显且相关性显著。以氧分压为横坐标,拉曼峰强为纵坐标,对pH为5.7,7.4和8.0时的血红蛋白在1 375和1 638cm-1峰的强度及其氧分压进行线性拟合,结果显示2处的拟合效果均良好,拉曼峰强度/氧分压的斜率关系表现为K8.0>K7.4>K5.7,表明pH越低血红蛋白的氧亲和力越低,越有利于向组织释放氧气。研究表明,应用拉曼光谱技术可检测血红蛋白与氧的亲和力,通过对血红蛋白-氧亲和力的量化检测,可以观察pH对血氧结合状态的影响,为血氧分压的监测以及氢离子和二氧化碳等血氧亲和力调节因子调节能力的研究提供新的研究方法与理论基础。The hemoglobin was extracted from the blood which was provided by the healthy volunteers and the impact of the pH on hemoglobin oxygen binding capacity was studied with microscopic Raman spectroscopy. The results indicated that： under the excitation light of 514.5 nm, with the reducing of the oxygen partial pressure （PO2）, the Raman peak intensity at 1 375, 1 562, 1 585 and 1 638 cm-1 of the control hemoglobin （pH 7. 4） reduced gradually, among which, the change of the 1 375 and 1 638 cm-1 were the most significant and had a good relevance with the PO2. The curves were plotted by regarding the PO2 as the x-axis and the Raman absolute intensity as the y-axis, and the relationship between hemoglobin Raman absolute intensity of the 1 375 and 1 638 cm-1 and their related PO2 levels when the pH was 5.7, 7. 4 and 8. 0 respectively were analyzed. The data was well linear fitted and the fitting equation was obtained. The relationship of the slope （Raman intensity/PO2 level） among them were K8. 0 〉K7.4 〉K5. 7, indicating that the lower the pH, the easier the release of the oxygen molecules. It was showed that the Raman spectroscopy technique could be used to detect the oxygen binding rate of hemoglobin quantitatively, and the effect of the PH on oxygen binding state of hemoglobin could be observed, which could provide a new method and make a foundation for the monitoring of the PO2 levels in the blood, as well as the research on the regulatory factors of the blood oxygen affinity, such as H＋ and CO2.

关 键 词：氢离子 pH值 血红蛋白 氧亲和力 氧分压 拉曼光谱 

分 类 号：Q632[生物学—生物物理学]