C57小鼠不同剂量自体血注射脑出血模型的建立与评价
发布时间：2019-06-12 19:27
【摘要】： 第一部分自体血输注小鼠脑出血模型成模评估 目的:利用二次注射法,建立不同剂量自体血输注的小鼠脑出血模型,评价造模动物的死亡率及神经功能改变。 方法:选取C57雄性小鼠104只,经随机分组,应用立体定向仪定位,通过二次注射法将5ul(N=28)、10ul(N=32)、20ul(N=30)不含抗凝剂的自体血注入基底节区,制作不同血肿量的脑出血模型,并以假手术组(N=14)为对照。术后统计各组死亡率,将各组存活小鼠再随机分为3个不同时间点的亚组,分别在术后第1、第2、第3天行前臂放置实验及转角实验。 结果:1.成功制作C57小鼠脑出血模型共86只。2.不同注射量的小鼠死亡率无统计学差异。3.各组间小鼠体重无统计学差异。4.前臂放置实验中5ul组、10ul组、20ul组及假手术组小鼠瘫痪侧与自身的健侧相比有明显统计学差异(P＜0.01);5ul组、10ul组、20ul组与假手术组瘫侧比较也显示明显差异(P＜0.01)。瘫侧结果中,5ul组-第2天为同组内最低值(P＜0.05);10ul组-第3天、20ul组-第3天及假手术组-第1天为各自组内最低值(P＞0.05);5ul-第1天与假手术组-第1天无统计学差异。5.转角实验中5ul组、10ul组、20ul组与假手术组LI结果相比有统计学差异(P＜0.05);5ul组-第2天、10ul组-第3天、20ul组-第3天、假手术组-第1天LI分别为各自组内最大值(P＞0.05);5ul-第3天与假手术组-第3天LI无统计学差异。 结论:5ul、10ul及20ul三种不同自体血量利用二次注射法均可形成具有明显偏侧神经功能缺损的小鼠脑出血模型,且损伤程度随注射量增加而明显。不同注射血量与小鼠死亡率无关。不同注射血量所致脑损伤最明显的时间点不同,分别为5ul-第2天、10ul-第3天、20ul-第3天及假手术组-第1天。5ul组模型创伤小,恢复快,特异性欠佳。 第二部分不同血肿量与神经功能变化的评估 目的:估算5ul组、10ul组及20ul组的血肿形成量,评估不同血肿量与神经功能变化的关系。 方法:前述成模小鼠19只,5ul组(N=6)、10ul组(N=7)、20ul组(N=6)各自随机分为3个不同时间点的亚组,分别在术后第1、第2、第3天行前臂放置实验及转角实验。取固定后脑组织冰冻切片,行H-E染色,划分血肿范围,估算血肿体积。 结果:1.血肿形成情况:5ul组、10ul组及20ul组小鼠均在基底节区形成局限性血肿;血肿实际大小分别为5ul组0.9868±0.2493mm~3,10ul组2.3254±0.3939mm~3,20ul组3.6465±0.6623mm~3。2.神经功能变化与血肿量的关系:随注射量增加三组形成的血肿量逐渐递增(P＜0.01);10ul组、20ul组瘫侧前臂放置均较5ul组差(P＜0.01),但10ul组与20ul组两者之间无统计学差异,与血肿量变化不一致。 结论:5ul组及10ul组均可形成与注射量接近的血肿,且神经功能变化与血肿形成量变化相关。20ul组由于操作时返流明显,形成的血肿量及神经功能变化不稳定。 第三部分脑水肿程度变化与神经功能变化的评估 目的:量化5ul组、10ul组、20ul组及假手术组的脑水肿情况,评估不同脑水肿程度与神经功能变化、不同血肿量与脑水肿程度的关系。 方法:前述成模小鼠67只,5ul组(N=18)、10ul组(N=19)、20ul组(N=18)及假手术组(N=12),各自随机分为3个不同时间点的亚组,分别在术后第1、第2、第3天行前臂放置实验及转角实验。取新鲜脑组织,利用干湿重法获得脑组织含水量。 结果:1.各组脑水肿情况:5ul组、10ul组、20ul组及假手术组患侧与自身的健侧相比有统计学差异(P＜0.05),10ul组、20ul组患侧均较假手术组有统计学差异(P＜0.05),5ul组患侧与假手术组无统计学差异,各组间健侧脑组织含水量结果无统计学差异;三组患侧最大值分别出现于5ul组-第2天、10ul组-第3天、20ul组-第3天,较自身对侧脑组织及假手术组患侧脑组织有统计学差异(P＜0.05),三者互相无统计学差异。2.神经功能变化与脑水肿程度的关系:5ul组、10ul组及20ul组的神经功能缺损结果较假手术组明显(P＜0.01),且程度逐渐递增;假手术组、5ul组、10ul(P＜0.01)组患侧脑组织含水量结果呈逐渐递增,但20ul组结果较10ul组结果下降(P＜0.05),与神经功能变化不一致。3.不同血肿量与脑水肿程度的关系:血肿量与患侧脑组织含水量在5ul组与10ul组有较好的递增对应关系,而20ul组的变化趋势相反。 结论:10ul组及20ul组患侧可形成较假手术组患侧更明显的脑水肿,而5ul组患侧脑水肿不明显。不同注射量的患侧脑水肿程度分别在5ul组-第2天、10ul组-第3天、20ul组-第3天、假手术组-第1天最明显。假手术组、5ul组、10ul组各自的神经功能缺损与患侧脑水肿程度呈良好的对应关系,5ul组与10ul组各自的血肿量与患侧脑水肿程度也呈良好的对应关系;而20ul组由于血肿形成量不稳定,其神经功能缺损及患侧脑组织含水量变化均未能形成对应增加。
[Abstract]:The evaluation of the model of cerebral hemorrhage in the first part of auto-transfusion of mice Objective: To establish the model of cerebral hemorrhage of mice with different dose of autologous blood infusion by the method of secondary injection and to evaluate the mortality and nerve work of the model animals. Methods:104 male mice were randomly divided,5 ul (N = 28),10 ul (N = 32),20 ul (N = 30) were injected into the basal ganglia area without anticoagulant, and the cerebral hemorrhage model of different hematomas was made. = 14) In the control group, the mortality of each group was counted after operation, and the surviving mice in each group were randomly divided into three sub-groups at different time points, and the forearm was placed on the first, second and third day after operation. experiment and corner The results were as follows:1. The C57 mice were successfully produced. A total of 86 rats with a cerebral hemorrhage model.2. Mice with different injection quantity There was no statistical difference in mortality.3. Small groups There was no statistical difference in the weight of the mice.4. There was a significant difference in the body weight of the control group (P <0.01) in the 5 ul group, the 10 ul group, the 20 ul group and the sham operation group in the forearm placement experiment. Significant difference (P <0.01). In the paralytic results, the lowest value (P <0.05) in the same group in the 5 ul group and the second day group (P <0.05); the lowest value in each group (P> 0.05) in the group of 10 ul-day 3, the 20 ul group and the third day and the sham operation group-day 1; and the 5 ul-day 1 and the sham operation group- There was no statistical difference in the first day.5 ul,10 ul, and 20 ul in the corner experiment were statistically different (P <0.05) compared to the results of the sham-operation group LI (P <0.05); the 5 ul group-day 2, the 10 ul group-day 3, the 20ul group-day 3, the sham-operation group-the first day LI, respectively, were within the respective groups Maximum (P> 0.05);5 ul-Day 3 and sham operation group- There was no statistical difference of LI in the third day. Conclusion:5 ul,10 ul and 20 ul of different autologous blood volume can form a model of cerebral hemorrhage with obvious side-side neurological deficit by using the secondary injection method. And the degree of damage is obviously increased with the increase of the injection amount. . The different injection volumes were not related to the mouse mortality. The most significant time points for brain damage due to different injection volumes were 5 ul-day 2,10 ul-day 3,20 ul-day 3, and sham-operation group-day 1.5 ul The model is small in trauma, fast in recovery and poor in specificity. The purpose of the second part of the assessment of the changes in the amount of hematoma and the neurological function was to estimate the 5 ul,10 ul and 20 ul groups. Methods:19,5 ul (N = 6),10 ul (N = 7) and 20 ul (N = 6) groups were randomly divided into 3 subgroups of different time points. In the first, second and third day of the operation, the forearm placement experiment and the angle experiment were performed. tissue freezing and cutting The results were as follows:1. The formation of the hematoma:1. The formation of the hematoma:1. The formation of the hematoma:5 ul,10 ul, and 20 ul of the mice had localized hematomas in the basal ganglia; the actual size of the hematoma was 0.9868 and 0.2493 mm ~ 3 in the 5ul group, 2.3254 and 0.39 in the 10ul group, respectively. The relationship between the changes of nerve function and the amount of hematoma in the group of 39mm ~ 3 and 20ul group was 3.6465 and 0.6623 mm ~ 3.2. The volume of hematoma formed by three groups was gradually increased with the increase of injection volume (P <0.01). There were 5 ul groups (P <0.01) in the group of 10 ul and 20ul group. There was no statistical difference between the group of 10 ul and the 20 ul group, and it was not consistent with the change in the volume of the hematoma. Conclusion: The hematoma with close to the injection amount can be formed in the 5 ul and the 10 ul groups, and the change of the function of the nerve function and the amount of hematoma formation The 20 ul group was formed due to the obvious regurgitation during operation. The volume of the hematoma and the changes of the neurological function were not stable. The purpose of the assessment of the change of the degree of cerebral edema in the third part and the changes of the neurological function was to quantify the 5 ul group, the 10 ul group and the 20 ul group. Methods:67,5 ul (N = 18),10 ul (N = 19),20 ul (N = 18) and sham-operated group (N = 18),20 ul (N = 18) and sham-operated group (N = 18) were evaluated. 12), each of which is divided into three sub-groups at different time points, and the sub-groups are divided into three sub-groups, Don't be post-op The results were as follows:1. The water content of brain tissue was obtained by dry-wet and dry-wet method in the first, second and third day. Results:1. The results showed that:1. There was a significant difference in the brain edema in each group (P <0.05). There was no statistical difference between the group and the sham operation group (P <0.05) in the group of 10 ul and 20 ul, there was no statistical difference between the group affected side and the sham operation group, and there was no statistical difference in the water content of the healthy side of the group. The maximum values of the three groups were found in the 5ul group and the second day respectively. Group 0 ul-Day 3, 20ul There was no statistical difference (P <0.05) between the group and the side of the group on the third day, and there was no statistical difference between the three groups.2. The relationship between the neurological function and the degree of cerebral edema: the results of the neurological deficit in the 5 ul group, the 10 ul group and the 20 ul group were more obvious than that in the sham operation group (P <0 .01), and gradually increased in degree; sham operation group,5 ul group,10 ul (P <0.01) group affected side brain group The results of water content increased gradually, but the results of the 20 ul group decreased significantly (P <0.05), which did not correspond to the change of the neurological function.3. The amount of different hematomas and the degree of cerebral edema The relationship between the amount of hematoma and the water content of the brain tissue in the affected side was better than that of the control group in the group of 5 ul and 10 ul, and the change of the 20ul group was the trend. Conclusion: There may be more obvious brain edema in the affected side of the group of 10 ul and 20 ul, while the brain edema in the 5 ul group is not obvious. The degree of brain edema at the affected side of the same injection was 5 ul group-day 2,10 ul group-day 3,20 ul group-day 3, and the sham operation group-day 1 was the most obvious. Corresponding relationship, the amount of hematoma in each of the 5 ul and the 10 ul groups and the cerebral edema in the affected side
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R743.34;R-332
本文编号：2498251
[Abstract]:The evaluation of the model of cerebral hemorrhage in the first part of auto-transfusion of mice Objective: To establish the model of cerebral hemorrhage of mice with different dose of autologous blood infusion by the method of secondary injection and to evaluate the mortality and nerve work of the model animals. Methods:104 male mice were randomly divided,5 ul (N = 28),10 ul (N = 32),20 ul (N = 30) were injected into the basal ganglia area without anticoagulant, and the cerebral hemorrhage model of different hematomas was made. = 14) In the control group, the mortality of each group was counted after operation, and the surviving mice in each group were randomly divided into three sub-groups at different time points, and the forearm was placed on the first, second and third day after operation. experiment and corner The results were as follows:1. The C57 mice were successfully produced. A total of 86 rats with a cerebral hemorrhage model.2. Mice with different injection quantity There was no statistical difference in mortality.3. Small groups There was no statistical difference in the weight of the mice.4. There was a significant difference in the body weight of the control group (P <0.01) in the 5 ul group, the 10 ul group, the 20 ul group and the sham operation group in the forearm placement experiment. Significant difference (P <0.01). In the paralytic results, the lowest value (P <0.05) in the same group in the 5 ul group and the second day group (P <0.05); the lowest value in each group (P> 0.05) in the group of 10 ul-day 3, the 20 ul group and the third day and the sham operation group-day 1; and the 5 ul-day 1 and the sham operation group- There was no statistical difference in the first day.5 ul,10 ul, and 20 ul in the corner experiment were statistically different (P <0.05) compared to the results of the sham-operation group LI (P <0.05); the 5 ul group-day 2, the 10 ul group-day 3, the 20ul group-day 3, the sham-operation group-the first day LI, respectively, were within the respective groups Maximum (P> 0.05);5 ul-Day 3 and sham operation group- There was no statistical difference of LI in the third day. Conclusion:5 ul,10 ul and 20 ul of different autologous blood volume can form a model of cerebral hemorrhage with obvious side-side neurological deficit by using the secondary injection method. And the degree of damage is obviously increased with the increase of the injection amount. . The different injection volumes were not related to the mouse mortality. The most significant time points for brain damage due to different injection volumes were 5 ul-day 2,10 ul-day 3,20 ul-day 3, and sham-operation group-day 1.5 ul The model is small in trauma, fast in recovery and poor in specificity. The purpose of the second part of the assessment of the changes in the amount of hematoma and the neurological function was to estimate the 5 ul,10 ul and 20 ul groups. Methods:19,5 ul (N = 6),10 ul (N = 7) and 20 ul (N = 6) groups were randomly divided into 3 subgroups of different time points. In the first, second and third day of the operation, the forearm placement experiment and the angle experiment were performed. tissue freezing and cutting The results were as follows:1. The formation of the hematoma:1. The formation of the hematoma:1. The formation of the hematoma:5 ul,10 ul, and 20 ul of the mice had localized hematomas in the basal ganglia; the actual size of the hematoma was 0.9868 and 0.2493 mm ~ 3 in the 5ul group, 2.3254 and 0.39 in the 10ul group, respectively. The relationship between the changes of nerve function and the amount of hematoma in the group of 39mm ~ 3 and 20ul group was 3.6465 and 0.6623 mm ~ 3.2. The volume of hematoma formed by three groups was gradually increased with the increase of injection volume (P <0.01). There were 5 ul groups (P <0.01) in the group of 10 ul and 20ul group. There was no statistical difference between the group of 10 ul and the 20 ul group, and it was not consistent with the change in the volume of the hematoma. Conclusion: The hematoma with close to the injection amount can be formed in the 5 ul and the 10 ul groups, and the change of the function of the nerve function and the amount of hematoma formation The 20 ul group was formed due to the obvious regurgitation during operation. The volume of the hematoma and the changes of the neurological function were not stable. The purpose of the assessment of the change of the degree of cerebral edema in the third part and the changes of the neurological function was to quantify the 5 ul group, the 10 ul group and the 20 ul group. Methods:67,5 ul (N = 18),10 ul (N = 19),20 ul (N = 18) and sham-operated group (N = 18),20 ul (N = 18) and sham-operated group (N = 18) were evaluated. 12), each of which is divided into three sub-groups at different time points, and the sub-groups are divided into three sub-groups, Don't be post-op The results were as follows:1. The water content of brain tissue was obtained by dry-wet and dry-wet method in the first, second and third day. Results:1. The results showed that:1. There was a significant difference in the brain edema in each group (P <0.05). There was no statistical difference between the group and the sham operation group (P <0.05) in the group of 10 ul and 20 ul, there was no statistical difference between the group affected side and the sham operation group, and there was no statistical difference in the water content of the healthy side of the group. The maximum values of the three groups were found in the 5ul group and the second day respectively. Group 0 ul-Day 3, 20ul There was no statistical difference (P <0.05) between the group and the side of the group on the third day, and there was no statistical difference between the three groups.2. The relationship between the neurological function and the degree of cerebral edema: the results of the neurological deficit in the 5 ul group, the 10 ul group and the 20 ul group were more obvious than that in the sham operation group (P <0 .01), and gradually increased in degree; sham operation group,5 ul group,10 ul (P <0.01) group affected side brain group The results of water content increased gradually, but the results of the 20 ul group decreased significantly (P <0.05), which did not correspond to the change of the neurological function.3. The amount of different hematomas and the degree of cerebral edema The relationship between the amount of hematoma and the water content of the brain tissue in the affected side was better than that of the control group in the group of 5 ul and 10 ul, and the change of the 20ul group was the trend. Conclusion: There may be more obvious brain edema in the affected side of the group of 10 ul and 20 ul, while the brain edema in the 5 ul group is not obvious. The degree of brain edema at the affected side of the same injection was 5 ul group-day 2,10 ul group-day 3,20 ul group-day 3, and the sham operation group-day 1 was the most obvious. Corresponding relationship, the amount of hematoma in each of the 5 ul and the 10 ul groups and the cerebral edema in the affected side
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R743.34;R-332
【共引文献】
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1 刘小利,罗祖明;脑出血的止血治疗[J];华西医学;2005年01期
2 邓鹏飞;赵伟佳;;重组活化凝血因子Ⅶ与超早期脑出血的止血治疗[J];医学综述;2008年07期
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