To investigate the effect of suspension pH on the structure and function of the hemoglobin in living crocodilian erythrocyte by laser confocal Raman spectroscopy. Methods: Living crocodilian blood was extracted used heparin as an anticoagulant. The isolated cells were putted into suspension solutions with pH 2.55, 3.02, 4.39, 5.50, 6.58, 7.13, 7.82, 8.31, 8.80 and 10.0 respectively. Then the Raman spectra was taken using a laser confocal Raman spectro-microscope to obtain information about the variation of the structure and oxygen carrying function of the hemoglobin as a function of pH. Results: 1. Hemoglobin in living crocodilian function normally between pH=3.02 to pH=8.80; 2. Ratio of oxyhemoglobin in hemoglobin of crocodilian erythrocyte is higher in pH=7.13 and pH=7.80. It reduces neither pH higher nor lower, 3.Crocodilian eiythrocyte adjusts the ratio of oxyhemoglobin to satisfy the different demand in different pH, in physiology pH condition it only need part of hemoglobin to satisfy the oxygen carrying capacity, in acidic condition, in need all hemoglobin to accomplish the oxygen carrying capacity, 4.Crocodilian erythrocyte has better acid resistance than alkali resistance. Conclusions: Croc odilian erythrocyte has a stronger ability to adapt to the environment and to carry oxygen.%目的:使用激光共焦拉曼光谱扫描术对不同pH环境下活态鳄鱼红细胞内血红蛋白结构功能的变化进行测定研究.以期揭示鳄鱼红细胞及其血红蛋白随不pH环境下的结构功能状态的变化规律.方法:鳄鱼血活体采集后用肝素抗凝,无菌分离出细胞后加入pH值分别为2.55、3.02、4.39、5.50、6.58、7.13、7.82、8.31、8.80、10.0的细胞缓冲溶液中,静置20 min后,使用拉曼光谱仪进行扫描获得红细胞拉曼谱,通过分析测定获得其血红蛋白的分子结构与携氧功能随pH值变化的情况.结果:1.鳄鱼红细胞血红蛋白在pH=3.02～pH=8.80均能保持正常的生理活性;2.鳄鱼红细胞中氧合蛋白占总蛋白百分比在pH=7.13和pH=7.80条件下最高,随着pH的升高或降低,红细胞中氧合蛋白占总血红蛋白的总量百分比均下降;3.鳄鱼红细胞通过调节携氧血红蛋白比例来适应不同pH值条件下的携氧需求,在正常pH条件下无需所有血红蛋白即可完成携氧功能,而在酸性条件下可以使用全部血红蛋白完成携氧功能;4.鳄鱼红细胞抗酸性能力比抗碱性能力强.结论:研究表明鳄鱼红细胞的血红蛋白具有比人类红细胞更强的环境适应能力和更强的携氧能力,对人造血的研究有指导作用.
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