Ⅳ胶原酶结合自体血构建脑出血性脑水肿大鼠实验模型  

Establishment of brain edema model of rats with intracerebral hemorrhage by Ⅳ collagenase combined with autologous blood

作　　者：赵晓勇[1] 张晓丽[1] 江向明[1] 

机构地区：[1]南方医科大学附属花都医院神经外科,广州510800

出　　处：《中华行为医学与脑科学杂志》2016年第10期891-895,共5页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：广东省科技厅科技项目（2015A030302003）;广东省中医药局科技项目（20151061）;广东省自然科学基金（2016A030313419）

摘　　要：目的探讨Ⅳ胶原酶结合自体血构建复合型脑出血性脑水肿大鼠模型，为脑水肿相关研究提供实验依据。方法180只SD大鼠随机均分6组：A组（Ⅳ胶原酶＋自体凝固血＋自体抗凝血）、B组（自体凝固血）、C组（Ⅳ胶原酶）、D组（自体抗凝血）、E组（空白对照）、F组（假手术）；A、B、C、D组建模，E组无处理，F组注射生理盐水；Longa／Bederson、肢体对称实验测评神经功能缺损；干湿重法测脑组织含水量、原子吸收光谱法测电解质K^＋、Na^＋、Ca^2＋；透射电镜（TEM）观察胶质细胞超微结构变化；免疫荧光法进行胶质细胞鉴定；FCM和TUNEL测胶质细胞凋亡；CCK-8和EDU测胶质细胞增殖；实时定量PCR和蛋白免疫印迹测定胶质细胞中基质金属蛋白酶-9（MMP-9）和水通道蛋白4（AQP4）mRNA和蛋白表达；划痕实验和侵袭小室（Transwell）测胶质细胞迁移及侵袭能力。结果与其他各组相比较，A组Longa／Bederson评分、脑组织含水量、电解质Na^＋最高，电解质K^＋、Ca^＋最少；A组胶质细胞凋亡最多（32．27±3．33），增殖最少（5．33±1．14），迁移（85．35±8．11）与侵袭（8．08±1．96）能力最低；A组中MMP．9和AQP4 mRNA（9．31±0．84；15．69±1．77）和蛋白（4．25±0.27；4.34±0．33）最高（P〈0．05）。结论Ⅳ胶原酶结合自体血构建的脑出血性脑水肿大鼠模型脑水肿标志物MMP-9和AQP4特异性更强，细胞学的功能性变化更显著。Objective To explore the model of IV combined with autologous blood to build a model of compound brain edema in rats, and provide experimental evidence for the related research of brain edema. Methods 180 SD rats were randomly divided into 6 groups： Group A （IV collagenase ＋ autologous coagulation blood ＋ autologous blood coagulation）, group B （autologous coagulation）, group C （IV eollagenase）, group D （ autologous anti coagulation） , E group （ blank control） , F group （ sham operation）.Group A, B, C, D were modeled, Group E was not given treatment, Group F was injected with normal saline.Neurological deficit was evalated by longa/Bederson and physical symmetry test score.The water content in brain tissue was detected by wet and dry weight method.K^＋, Na^＋, Ca^2＋ were detected by atomic absorption spectrometry.The uhrastructural changes of glial cells were observed by transmission electron microscope （ TEM ）. Glial cells were identified by immunofluorescence assay.FCM and TUNEL tested the apoptosis of glial cells.CCK-8 and EDU tested the proliferation of glial cells.Real time quantitative polymerase chain reaction and Western blot were used to measure matrix metalloproteinase-9（MMP-9） in glial cells and water channel protein 4（AQP4） mRNA and protein expression.The scratch test and Transwell assayed glial cell migration and invasion.Results Comparison among groups ： Longa/Bederson score of Neurologic impairment, Brain tissue water content, electrolyte Na^＋ was the highest level of group A, electrolyte K^＋, Ca^2＋ at least.The apoptosis of group A was the most （32.27±3.33 ）, the least proliferation （5.33±1.14）, migration and invasion ability were the wea- kest（85.35±8.11;8.08±1.96）; mRNA（9.31±0.84;15.69±1.77） and protein（4.25±0.27;4.34±0.33） of MMP-9 and AQP4 were the most significant in group A （P〈 0.05 ）. Conclusion IV collagenase combined with autologous blood to build a model of brain edema animal model makes the brain edem

关 键 词：脑出血 脑水肿 大鼠 动物模型 基质金属蛋白酶-9 水通道蛋白4 

分 类 号：R743.34[医药卫生—神经病学与精神病学] R-332[医药卫生—临床医学]