温度对人红细胞及血红蛋白结构功能的影响
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摘要
温度作为血红蛋白和红细胞的一个重要变构因子，其微小变化即可引起从血红蛋白的分子结构、浓度、功能以至红细胞的形态、结构、功能的显著改变。本文采用显微激光共焦拉曼散射技术、动态光散射技术、紫外—可见光谱技术以及多维显微图像分析技术等多种手段，从人体可能发生高温的温度特征及持续时间有时会较长的实际情况出发，从形态、结构及功能三个方面对血红蛋白和红细胞随温度的改变所对应的变化情况进行研究测定。本文发现：
1．离体血红蛋白在高温(39℃-41℃)或低温(4℃-30℃)情况下，只要达到一定的作用时间，就会发生团聚。39℃-41℃团聚体的粒径在100nm到200nm间，且随温度的升高与作用时间的延长而增大。
2．离体血红蛋白在39℃-41℃高温作用下，其二级结构、空间结构会发生变化，主要表现在有序结构明显减少，无规卷曲增加、血红蛋白卟啉环丢失或松散、疏水性氨基酸残基暴露在外，血红蛋白的携氧能力也随加热温度的升高和加热时间的延长呈下降趋势。
3．高温下血红蛋白的团聚及其分子结构与功能的变化是不可逆行为。而低温作用下血红蛋白的团聚是物理团聚，是可逆的过程，且其分子结构以及功能没有发生变化。
4．高温下孵育的红细胞其形态从正常双凹形转变成棘形，且随加热温度的升高与加热时间的延长红细胞的畸变率升高；且胞内血红蛋白分子结构与功能也相应发生了变化。
本文的研究填补了有关温度对红细胞及其血红蛋白结构和功能的影响作用随作用时间的变化关系，对于阐明红细胞及血红蛋白在有关生理、病理状态下的结构与功能的变化，以及其临床表现与有关防治对策都有重要的指导意义。
As a significant factor, temperature would affect the structures and functions of humanerythrocyte and hemoglobin. Because of the fact that human may have fever and sometimes itwould keep on a long time, so in this thesis, the effect of temperature on the structure andfunction of erythrocyte and hemoglobin was studied by using some advanced techniques likeConfocal laser Raman microscopy, dynamic light scattering, ultraviolet-visible absorptionspectrum, and multi-dimensional micro-imaging. According to the experimental results, we havethe following conclusion:
1. Ex vivo hemoglobin would aggregate at the higher temperature (from 39 degree to 41degree) or lower temperature (from 4 degree to 30 degree) than the normal bodytemperature. It is found that the diameter of the aggregation would increase from100nm to 200nm when the temperature rises from 39 degree to 41 degree.
2. The protein structure of the ex vivo hemoglobin would change by the action of highertemperature, and exhibits obvious decrease in its nonrandom structure, increase withandom coil, loosened porphyrin in the hemoglobin, exposure of the hydrophobic aminoacid. In addition, the oxygen carrying capacity of the red blood cell decreases withincreasing temperature and warm-up time.
3. It is found that at higher temperature, once upon the aggregates are formed; it wouldnot disaggregate to normal tetramer by lower down temperature. However, theaggregates formed at lower temperature would disaggregate when they are put back tothe normal body temperature. In addition, the protein's structure and function are notinfluenced.
4. Red blood cell would change its shape from biconcave disc to echinocyte at highertemperature. The distortion rate of the erythrocyte at higher temperature increase withtemperature and the warm-up time, and the molecular structure and function of theintercellular hemoglobin are also changed.
This work not only reveals the effect of temperature on the structure and functions oferythrocyte and ex vivo hemoglobin, as a function of the warm-up time, but also elucidates the possible variation of the erythrocyte and hemoglobin under different pathologic states, isbelieved to be significant for the prevention and treatment of some diseases.
1．离体血红蛋白在高温(39℃-41℃)或低温(4℃-30℃)情况下，只要达到一定的作用时间，就会发生团聚。39℃-41℃团聚体的粒径在100nm到200nm间，且随温度的升高与作用时间的延长而增大。
2．离体血红蛋白在39℃-41℃高温作用下，其二级结构、空间结构会发生变化，主要表现在有序结构明显减少，无规卷曲增加、血红蛋白卟啉环丢失或松散、疏水性氨基酸残基暴露在外，血红蛋白的携氧能力也随加热温度的升高和加热时间的延长呈下降趋势。
3．高温下血红蛋白的团聚及其分子结构与功能的变化是不可逆行为。而低温作用下血红蛋白的团聚是物理团聚，是可逆的过程，且其分子结构以及功能没有发生变化。
4．高温下孵育的红细胞其形态从正常双凹形转变成棘形，且随加热温度的升高与加热时间的延长红细胞的畸变率升高；且胞内血红蛋白分子结构与功能也相应发生了变化。
本文的研究填补了有关温度对红细胞及其血红蛋白结构和功能的影响作用随作用时间的变化关系，对于阐明红细胞及血红蛋白在有关生理、病理状态下的结构与功能的变化，以及其临床表现与有关防治对策都有重要的指导意义。
As a significant factor, temperature would affect the structures and functions of humanerythrocyte and hemoglobin. Because of the fact that human may have fever and sometimes itwould keep on a long time, so in this thesis, the effect of temperature on the structure andfunction of erythrocyte and hemoglobin was studied by using some advanced techniques likeConfocal laser Raman microscopy, dynamic light scattering, ultraviolet-visible absorptionspectrum, and multi-dimensional micro-imaging. According to the experimental results, we havethe following conclusion:
1. Ex vivo hemoglobin would aggregate at the higher temperature (from 39 degree to 41degree) or lower temperature (from 4 degree to 30 degree) than the normal bodytemperature. It is found that the diameter of the aggregation would increase from100nm to 200nm when the temperature rises from 39 degree to 41 degree.
2. The protein structure of the ex vivo hemoglobin would change by the action of highertemperature, and exhibits obvious decrease in its nonrandom structure, increase withandom coil, loosened porphyrin in the hemoglobin, exposure of the hydrophobic aminoacid. In addition, the oxygen carrying capacity of the red blood cell decreases withincreasing temperature and warm-up time.
3. It is found that at higher temperature, once upon the aggregates are formed; it wouldnot disaggregate to normal tetramer by lower down temperature. However, theaggregates formed at lower temperature would disaggregate when they are put back tothe normal body temperature. In addition, the protein's structure and function are notinfluenced.
4. Red blood cell would change its shape from biconcave disc to echinocyte at highertemperature. The distortion rate of the erythrocyte at higher temperature increase withtemperature and the warm-up time, and the molecular structure and function of theintercellular hemoglobin are also changed.
This work not only reveals the effect of temperature on the structure and functions oferythrocyte and ex vivo hemoglobin, as a function of the warm-up time, but also elucidates the possible variation of the erythrocyte and hemoglobin under different pathologic states, isbelieved to be significant for the prevention and treatment of some diseases.
引文
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