三种血瘀模型红细胞膜生物学差异性及寒冷刺激血瘀模型方证相关实验研究
发布时间：2019-05-24 20:35
【摘要】：目的：从红细胞膜分子层次探讨大鼠寒冷刺激血瘀模型、饥饿加力竭游泳血瘀模型、声光电复合刺激加钳尾激惹血瘀模型在血液流变学方面的生物学差异；应用少腹逐瘀汤、补阳还五汤、丹参饮反证寒冷刺激血瘀模型红细胞膜生物学特征 方法：以大鼠为研究对象。(1)用冰水浸泡模拟寒冷刺激而致血瘀模型、以饥饿加力竭游泳模拟气虚血瘀模型、以不可预期的声光电复合刺激加钳尾激惹模拟惊恐、愤怒过度而致气滞血瘀模型。持续刺激直至2周后各组大鼠分别出现寒凝血瘀、气虚血瘀、气滞血瘀表征,颈动脉取血测血液黏度、红细胞变形性、血沉K值、凝血四项、血浆胆固醇及甘油三酯含量；取血制备红细胞膜,检测红细胞膜Na+-K+-ATP酶活力、唾液酸含量、巯基含量、胆固醇含量、SOD活性、MDA含量。(2)寒冷刺激血瘀模型大鼠分别灌胃给予高、低剂量少腹逐瘀汤,高、低剂量补阳还五汤及高、低剂量丹参饮,连续2周,颈动脉取血,除检测上述指标外,以激光共聚焦显微镜观察各组大鼠红细胞膜骨架结构,并检测红细胞膜蛋白组成及百分含量。 结果：(1)与空白对照比较,寒冷刺激血瘀模型组全血黏度、血沉K值、FIB含量均显著增高,红细胞变形能力明显降低,APTT明显缩短,红细胞膜Na+-K+-ATP酶活力、SOD活性显著下降,唾液酸含量显著降低(P0.05或P0.01)；饥饿加力竭游泳血瘀模型高切变率全血黏度、红细胞变形性降低,APTT及PT明显缩短,血浆胆固醇含量下降,红细胞膜红细胞膜Na+-K+-ATP酶活力及SOD活性显著降低(P0.05或P0.01)；不可预期声光电刺激加钳尾激惹血瘀大鼠APTT明显缩短,红细胞膜Na+-K+-ATP酶活力及SOD活性显著下降、唾液酸含量显著降低,MDA含量增加(P0.05或P0.01)。(2)与寒冷刺激血瘀模型组比较,少腹逐瘀汤、补阳还五汤、丹参饮均可显著降低低切变率全血黏度,在高切变率下全血黏度少腹逐瘀汤降低效果最好；三方均能显著升高红细胞变形指数显著降低FIB含量,增加红细胞膜上唾液酸的含量及SOD酶活力(P0.05或P0.01)；丹参饮可显著降低K值(P0.05),升高红细胞膜带1蛋白百分含量；少腹逐瘀汤可显著纠正APTT缩短及降低MDA含量,提高Na+-K+-ATP酶活力,明显改善F-actin解聚状态,升高红细胞膜上带1蛋白、带2蛋白、带3蛋白百分含量(P0.05或P0.01)。 结论：(1)宏观血液流变学方面：寒冷刺激血瘀模型血液流变学呈高流变性特征；饥饿加力竭游泳血瘀模型血液流变学呈现低流变特点；声光电复合刺激加钳尾激惹血瘀模型除APTT明显缩短,其余宏观血液流变学指标变化不明显；(2)三种血瘀模型均表现为红细胞膜Na+-K+-ATP酶活性、SOD活性降低；寒冷刺激和声光电复合刺激血瘀模型红细胞膜唾液酸含量减少；(3)补阳还五汤、少腹逐瘀汤、丹参饮三方可纠正寒冷刺激血瘀模型全血黏度增高、红细胞变形性降低、聚集性增强、纤维蛋白原含量增加及膜骨架构象异常；纠正APTT缩短、提高膜Na+-K+-ATP酶活性、减少膜MDA含量方面,少腹逐瘀汤优于其他两方；少腹逐瘀汤可纠正寒冷刺激血瘀模型红细胞膜带1蛋白、带2蛋白、带3蛋白百分含量异常,丹参饮作用于膜带1蛋白、带7蛋白。
[Abstract]:Objective: To study the biological difference of the blood stasis model of the rat cold-stimulated blood stasis model, the starvation and the exhaustive swimming and blood stasis model and the acoustic-photoelectrical composite stimulation of the blood stasis model in the blood rheology from the molecular level of the red cell membrane. The biological characteristics of red cell membrane in the model of cold stimulation and blood stasis of Danshen Yin Methods: The rats were used as the research group. The method comprises the following steps of: (1) soaking in ice water to simulate the blood stasis model due to cold stimulation, and simulating the blood stasis model by using the hunger and exhausted swimming to simulate the blood stasis model, Type: After 2 weeks of continuous stimulation, the rats in each group had cold and blood stasis, Qi deficiency and blood stasis, qi stagnation and blood stasis, blood viscosity, erythrocyte deformability, erythrocyte sedimentation rate, blood coagulation, blood coagulation, plasma cholesterol and triglyceride content, and blood-taking preparation of red blood cells. The membrane was used to detect the activity of Na +-K +-ATPase in erythrocyte membrane, the content of sialic acid, the content of sodium peroxide, the content of cholesterol, the activity of SOD and the content of MDA. (2) In cold-stimulated and blood-stasis model rats, the rats were given high, low-dose, low-dose, no-blood-stasis decoction, high-dose, low-dose Buyang Huanwu decoction and high-dose, low-dose red-rooted salvia, for 2 weeks, the carotid artery was taken for blood, and the above-mentioned indexes were detected. In addition, the structure of red cell membrane was observed by laser confocal microscope and the composition and percentage of red blood cell membrane protein were detected. Results: (1) Compared with the blank control, the whole blood viscosity, the erythrocyte sedimentation rate and the FIB content of the model group were significantly higher than that of the blank control, and the erythrocyte deformability was significantly reduced, the APTT was obviously shortened, the Na +-K +-ATPase activity and the activity of SOD in the erythrocyte membrane were significantly shortened. There was a significant decrease in the content of sialic acid (P0.05 or P0.01), the high shear rate of the whole blood viscosity, the decrease of erythrocyte deformability, the decrease of APTT and PT, the decrease of plasma cholesterol, the decrease of erythrocyte membrane Na +-K +-ATPase activity and the activity of SOD (P0.05 or P0). The activity of Na +-K +-ATPase and the activity of SOD and the content of sialic acid decreased significantly, and the content of MDA increased (P0.05 or P0). (1). (2) Compared with the cold-stimulated blood stasis model group, the whole blood viscosity of the whole blood with low shear rate can be obviously reduced, the whole blood viscosity of the whole blood is reduced, the blood viscosity of the whole blood is reduced and the effect of the blood stasis decoction is reduced under the high shear rate, The results showed that the red blood cell deformation index significantly decreased the content of the FIB, increased the content of the sialic acid on the erythrocyte membrane and the activity of the SOD (P0.05 or P0.01). The content of APTT and the content of MDA, Na +-K +-ATPase activity were significantly improved, the activity of Na +-K +-ATPase was improved, the depolymerized state of F-actin was obviously improved, and 1 protein,2-protein and 3-protein in the erythrocyte membrane were increased (P0.05 or P0). 01). Conclusion: (1) The blood rheology in the model of blood rheology in cold-stimulated blood-stasis model is characterized by high rheological property, and the blood rheology of the model is characterized by low rheology. (2) The activity of Na +-K +-ATPase and the decrease of SOD activity of erythrocyte membrane Na +-K +-ATPase were observed in the three blood stasis models. The blood viscosity of the whole blood of the cold-stimulated blood-stasis model can be corrected by the three-way of the lower abdomen and the blood-stasis decoction, the deformability of the red blood cell is reduced, the aggregation is enhanced, the content of the fibrinogen is increased, and the conformation of the membrane skeleton is abnormal; the APTT is shortened, the Na +-K +-ATPase activity of the membrane is improved, the membrane MD is reduced, In the aspect of content A, the lower abdomen and the blood stasis decoction is superior to the other two; the lower abdomen and the blood stasis decoction can correct the red cell membrane band 1 protein, the 2 protein and the 3 protein content of the red cell membrane of the cold stimulation blood stasis model, the red sage root drink is applied to the membrane band 1 protein,
【学位授予单位】：黑龙江中医药大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R-332
本文编号：2485165
[Abstract]:Objective: To study the biological difference of the blood stasis model of the rat cold-stimulated blood stasis model, the starvation and the exhaustive swimming and blood stasis model and the acoustic-photoelectrical composite stimulation of the blood stasis model in the blood rheology from the molecular level of the red cell membrane. The biological characteristics of red cell membrane in the model of cold stimulation and blood stasis of Danshen Yin Methods: The rats were used as the research group. The method comprises the following steps of: (1) soaking in ice water to simulate the blood stasis model due to cold stimulation, and simulating the blood stasis model by using the hunger and exhausted swimming to simulate the blood stasis model, Type: After 2 weeks of continuous stimulation, the rats in each group had cold and blood stasis, Qi deficiency and blood stasis, qi stagnation and blood stasis, blood viscosity, erythrocyte deformability, erythrocyte sedimentation rate, blood coagulation, blood coagulation, plasma cholesterol and triglyceride content, and blood-taking preparation of red blood cells. The membrane was used to detect the activity of Na +-K +-ATPase in erythrocyte membrane, the content of sialic acid, the content of sodium peroxide, the content of cholesterol, the activity of SOD and the content of MDA. (2) In cold-stimulated and blood-stasis model rats, the rats were given high, low-dose, low-dose, no-blood-stasis decoction, high-dose, low-dose Buyang Huanwu decoction and high-dose, low-dose red-rooted salvia, for 2 weeks, the carotid artery was taken for blood, and the above-mentioned indexes were detected. In addition, the structure of red cell membrane was observed by laser confocal microscope and the composition and percentage of red blood cell membrane protein were detected. Results: (1) Compared with the blank control, the whole blood viscosity, the erythrocyte sedimentation rate and the FIB content of the model group were significantly higher than that of the blank control, and the erythrocyte deformability was significantly reduced, the APTT was obviously shortened, the Na +-K +-ATPase activity and the activity of SOD in the erythrocyte membrane were significantly shortened. There was a significant decrease in the content of sialic acid (P0.05 or P0.01), the high shear rate of the whole blood viscosity, the decrease of erythrocyte deformability, the decrease of APTT and PT, the decrease of plasma cholesterol, the decrease of erythrocyte membrane Na +-K +-ATPase activity and the activity of SOD (P0.05 or P0). The activity of Na +-K +-ATPase and the activity of SOD and the content of sialic acid decreased significantly, and the content of MDA increased (P0.05 or P0). (1). (2) Compared with the cold-stimulated blood stasis model group, the whole blood viscosity of the whole blood with low shear rate can be obviously reduced, the whole blood viscosity of the whole blood is reduced, the blood viscosity of the whole blood is reduced and the effect of the blood stasis decoction is reduced under the high shear rate, The results showed that the red blood cell deformation index significantly decreased the content of the FIB, increased the content of the sialic acid on the erythrocyte membrane and the activity of the SOD (P0.05 or P0.01). The content of APTT and the content of MDA, Na +-K +-ATPase activity were significantly improved, the activity of Na +-K +-ATPase was improved, the depolymerized state of F-actin was obviously improved, and 1 protein,2-protein and 3-protein in the erythrocyte membrane were increased (P0.05 or P0). 01). Conclusion: (1) The blood rheology in the model of blood rheology in cold-stimulated blood-stasis model is characterized by high rheological property, and the blood rheology of the model is characterized by low rheology. (2) The activity of Na +-K +-ATPase and the decrease of SOD activity of erythrocyte membrane Na +-K +-ATPase were observed in the three blood stasis models. The blood viscosity of the whole blood of the cold-stimulated blood-stasis model can be corrected by the three-way of the lower abdomen and the blood-stasis decoction, the deformability of the red blood cell is reduced, the aggregation is enhanced, the content of the fibrinogen is increased, and the conformation of the membrane skeleton is abnormal; the APTT is shortened, the Na +-K +-ATPase activity of the membrane is improved, the membrane MD is reduced, In the aspect of content A, the lower abdomen and the blood stasis decoction is superior to the other two; the lower abdomen and the blood stasis decoction can correct the red cell membrane band 1 protein, the 2 protein and the 3 protein content of the red cell membrane of the cold stimulation blood stasis model, the red sage root drink is applied to the membrane band 1 protein,
【学位授予单位】：黑龙江中医药大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R-332
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