血液粘度增高模型动物及其血清致脑细胞毒性的实验研究

发布时间：2018-06-27 10:47

本文选题：血粘度增高动物模型 + 脑缺氧　；参考：《兰州大学》2006年硕士论文

【摘要】： 目的：1．探讨血粘度增高模型大鼠血液粘度与动脉血氧含量的相关性。2．用模型动物血清干预脑细胞，观察细胞凋亡及细胞内游离钙离子([Ca~(2+)]i)浓度变化，探讨模型动物血清中存在的致脑细胞毒性因素。方法：1．采用高分子右旋糖酐(dextran，DT)法建立血粘度增高大鼠模型，用血液粘度仪、血液分析仪检测血常规及血液流变学指标的变化。 2．用i-STAT临床血液分析系统(EG7~+血气片)检测造模大鼠颈内动、静脉血氧饱和度及血氧分压的变化。 3．以Ca~(2+)特异性荧光探针Flou-3／AM为荧光指示剂，采用不同时间造模大鼠血清干预培养的大鼠原代脑细胞，在激光共聚焦显微镜下分时间段实时动态观察[Ca~(2+)]i的变化。 4．以吖啶橙(acridine orange，AO)和碘化丙锭(propidium iodide，PI)为染色剂，在激光共聚焦显微镜下分时间段观察经造模5d大鼠血清培养的原代脑细胞凋亡和死亡情况。结果：1．造模时间与全血粘度和血浆粘度呈正相关，与血液有形成份持续减少呈负相关。 2．造模大鼠全血粘度和血浆粘度与动脉血氧含量(CaO_2)、静脉血氧饱和度(SjvO_2)、静脉血氧分压(PvO_2)及静脉血氧含量(CjvO_2)呈负相关，而与动脉血氧饱和度(SaO_2)、动脉血氧分压(PaO_2)无相关性。 3．与正常对照比较，造模20min血清干预导致脑细胞[Ca~(2+)]i出现钙波动；造模5d血清干预导致脑细胞[Ca~(2+)]i出现持续性钙超载。 4．造模5d血清干预脑细胞，，在30min内出现凋亡和死亡细胞，且凋亡和死亡程度随干预时间的延长而加重。结论：1．DT制作血粘度增高模型动物方法简单易行，重复率高是理想的慢性缺氧缺血模型动物。 2．CaO_2可作为DT制作血粘度增高模型动物组织缺氧程度的评估指标。 3．造模5d动物血清中存在致脑细胞损伤因素，是何种成份起作用有待进一步研究。
[Abstract]:Objective 1. To investigate the correlation between blood viscosity and arterial blood oxygen content in rats with increased blood viscosity. 2. To observe apoptosis and the concentration of intracellular free calcium ([Ca ~ (2)] I) in brain cells with model animal serum. To investigate the factors of brain cytotoxicity in the serum of model animals. Methods 1. The rat model of increased blood viscosity was established by dextran (DT) method, and blood viscometer was used. The changes of blood routine and hemorheological indexes were detected by blood analyzer. 2. The internal motion of the model rats was detected by i-STAT clinical blood analysis system (EG7 ~ XG). The changes of oxygen saturation and partial pressure of oxygen in vein. 3. The primary brain cells of rats were cultured with different time serum of model rats with Ca ~ (2) specific fluorescence probe Flou-3 / AM as fluorescent indicator. The changes of [Ca ~ (2)] _ I were observed in real time under laser confocal microscope. (4) acridine orange (acridine orangeo) and propidium iodide (Pi) were used as chromatin. The apoptosis and death of primary brain cells cultured in rat serum for 5 days were observed under laser confocal microscope. Results (1) the modeling time was positively correlated with the whole blood viscosity and plasma viscosity. The whole blood viscosity and plasma viscosity were negatively correlated with the arterial oxygen content (CaO2), the venous oxygen saturation (SjvO _ 2), the venous oxygen partial pressure (PvO _ 2) and the venous blood oxygen content (CjvO _ 2) in the model rats. However, there was no correlation with Sao _ 2 and Pao _ 2. 3. Compared with normal control, serum intervention of model 20min resulted in calcium fluctuation of [Ca ~ (2)] _ I in brain cells. The serum intervention for 5 days resulted in persistent calcium overload in brain cells. 4. After 5 days of model making, there were apoptosis and death cells in 30min, and the degree of apoptosis and death increased with the prolongation of intervention time. Conclusion 1. The method of making animal model with increased blood viscosity is simple and feasible. High repetition rate is an ideal chronic hypoxic ischemia model animal. 2. CaOs _ 2 can be used as an index to evaluate the hypoxic degree of model animal with increased blood viscosity of DT. 3. Modeling for 5 days There are brain cell injury factors in animal serum. What kind of composition works needs further study.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R363

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