C57小鼠不同劑量自體血注射腦出血模型的建立與評價
發(fā)布時間：2019-06-12 19:27
【摘要】： 第一部分自體血輸注小鼠腦出血模型成模評估 目的:利用二次注射法,建立不同劑量自體血輸注的小鼠腦出血模型,評價造模動物的死亡率及神經(jīng)功能改變。 方法:選取C57雄性小鼠104只,經(jīng)隨機(jī)分組,應(yīng)用立體定向儀定位,通過二次注射法將5ul(N=28)、10ul(N=32)、20ul(N=30)不含抗凝劑的自體血注入基底節(jié)區(qū),制作不同血腫量的腦出血模型,并以假手術(shù)組(N=14)為對照。術(shù)后統(tǒng)計各組死亡率,將各組存活小鼠再隨機(jī)分為3個不同時間點(diǎn)的亞組,分別在術(shù)后第1、第2、第3天行前臂放置實(shí)驗(yàn)及轉(zhuǎn)角實(shí)驗(yàn)。 結(jié)果:1.成功制作C57小鼠腦出血模型共86只。2.不同注射量的小鼠死亡率無統(tǒng)計學(xué)差異。3.各組間小鼠體重?zé)o統(tǒng)計學(xué)差異。4.前臂放置實(shí)驗(yàn)中5ul組、10ul組、20ul組及假手術(shù)組小鼠癱瘓側(cè)與自身的健側(cè)相比有明顯統(tǒng)計學(xué)差異(P＜0.01);5ul組、10ul組、20ul組與假手術(shù)組癱側(cè)比較也顯示明顯差異(P＜0.01)。癱側(cè)結(jié)果中,5ul組-第2天為同組內(nèi)最低值(P＜0.05);10ul組-第3天、20ul組-第3天及假手術(shù)組-第1天為各自組內(nèi)最低值(P＞0.05);5ul-第1天與假手術(shù)組-第1天無統(tǒng)計學(xué)差異。5.轉(zhuǎn)角實(shí)驗(yàn)中5ul組、10ul組、20ul組與假手術(shù)組LI結(jié)果相比有統(tǒng)計學(xué)差異(P＜0.05);5ul組-第2天、10ul組-第3天、20ul組-第3天、假手術(shù)組-第1天LI分別為各自組內(nèi)最大值(P＞0.05);5ul-第3天與假手術(shù)組-第3天LI無統(tǒng)計學(xué)差異。 結(jié)論:5ul、10ul及20ul三種不同自體血量利用二次注射法均可形成具有明顯偏側(cè)神經(jīng)功能缺損的小鼠腦出血模型,且損傷程度隨注射量增加而明顯。不同注射血量與小鼠死亡率無關(guān)。不同注射血量所致腦損傷最明顯的時間點(diǎn)不同,分別為5ul-第2天、10ul-第3天、20ul-第3天及假手術(shù)組-第1天。5ul組模型創(chuàng)傷小,恢復(fù)快,特異性欠佳。 第二部分不同血腫量與神經(jīng)功能變化的評估 目的:估算5ul組、10ul組及20ul組的血腫形成量,評估不同血腫量與神經(jīng)功能變化的關(guān)系。 方法:前述成模小鼠19只,5ul組(N=6)、10ul組(N=7)、20ul組(N=6)各自隨機(jī)分為3個不同時間點(diǎn)的亞組,分別在術(shù)后第1、第2、第3天行前臂放置實(shí)驗(yàn)及轉(zhuǎn)角實(shí)驗(yàn)。取固定后腦組織冰凍切片,行H-E染色,劃分血腫范圍,估算血腫體積。 結(jié)果:1.血腫形成情況:5ul組、10ul組及20ul組小鼠均在基底節(jié)區(qū)形成局限性血腫;血腫實(shí)際大小分別為5ul組0.9868±0.2493mm~3,10ul組2.3254±0.3939mm~3,20ul組3.6465±0.6623mm~3。2.神經(jīng)功能變化與血腫量的關(guān)系:隨注射量增加三組形成的血腫量逐漸遞增(P＜0.01);10ul組、20ul組癱側(cè)前臂放置均較5ul組差(P＜0.01),但10ul組與20ul組兩者之間無統(tǒng)計學(xué)差異,與血腫量變化不一致。 結(jié)論:5ul組及10ul組均可形成與注射量接近的血腫,且神經(jīng)功能變化與血腫形成量變化相關(guān)。20ul組由于操作時返流明顯,形成的血腫量及神經(jīng)功能變化不穩(wěn)定。 第三部分腦水腫程度變化與神經(jīng)功能變化的評估 目的:量化5ul組、10ul組、20ul組及假手術(shù)組的腦水腫情況,評估不同腦水腫程度與神經(jīng)功能變化、不同血腫量與腦水腫程度的關(guān)系。 方法:前述成模小鼠67只,5ul組(N=18)、10ul組(N=19)、20ul組(N=18)及假手術(shù)組(N=12),各自隨機(jī)分為3個不同時間點(diǎn)的亞組,分別在術(shù)后第1、第2、第3天行前臂放置實(shí)驗(yàn)及轉(zhuǎn)角實(shí)驗(yàn)。取新鮮腦組織,利用干濕重法獲得腦組織含水量。 結(jié)果:1.各組腦水腫情況:5ul組、10ul組、20ul組及假手術(shù)組患側(cè)與自身的健側(cè)相比有統(tǒng)計學(xué)差異(P＜0.05),10ul組、20ul組患側(cè)均較假手術(shù)組有統(tǒng)計學(xué)差異(P＜0.05),5ul組患側(cè)與假手術(shù)組無統(tǒng)計學(xué)差異,各組間健側(cè)腦組織含水量結(jié)果無統(tǒng)計學(xué)差異;三組患側(cè)最大值分別出現(xiàn)于5ul組-第2天、10ul組-第3天、20ul組-第3天,較自身對側(cè)腦組織及假手術(shù)組患側(cè)腦組織有統(tǒng)計學(xué)差異(P＜0.05),三者互相無統(tǒng)計學(xué)差異。2.神經(jīng)功能變化與腦水腫程度的關(guān)系:5ul組、10ul組及20ul組的神經(jīng)功能缺損結(jié)果較假手術(shù)組明顯(P＜0.01),且程度逐漸遞增;假手術(shù)組、5ul組、10ul(P＜0.01)組患側(cè)腦組織含水量結(jié)果呈逐漸遞增,但20ul組結(jié)果較10ul組結(jié)果下降(P＜0.05),與神經(jīng)功能變化不一致。3.不同血腫量與腦水腫程度的關(guān)系:血腫量與患側(cè)腦組織含水量在5ul組與10ul組有較好的遞增對應(yīng)關(guān)系,而20ul組的變化趨勢相反。 結(jié)論:10ul組及20ul組患側(cè)可形成較假手術(shù)組患側(cè)更明顯的腦水腫,而5ul組患側(cè)腦水腫不明顯。不同注射量的患側(cè)腦水腫程度分別在5ul組-第2天、10ul組-第3天、20ul組-第3天、假手術(shù)組-第1天最明顯。假手術(shù)組、5ul組、10ul組各自的神經(jīng)功能缺損與患側(cè)腦水腫程度呈良好的對應(yīng)關(guān)系,5ul組與10ul組各自的血腫量與患側(cè)腦水腫程度也呈良好的對應(yīng)關(guān)系;而20ul組由于血腫形成量不穩(wěn)定,其神經(jīng)功能缺損及患側(cè)腦組織含水量變化均未能形成對應(yīng)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
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R743.34;R-332
本文編號：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
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R743.34;R-332
【共引文獻(xiàn)】
相關(guān)期刊論文 前3條
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1 鄧鵬飛;554例腦出血患者預(yù)后相關(guān)因素分析[D];廣西醫(yī)科大學(xué);2008年
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