载脂蛋白E短肽COG1410对蛛网膜下腔出血后早期脑水肿的影响

发布时间：2018-01-03 06:18

本文关键词：载脂蛋白E短肽COG1410对蛛网膜下腔出血后早期脑水肿的影响　出处：《西南医科大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的:探讨载脂蛋白E短肽COG1410对小鼠实验性蛛网膜下腔出血(subarachnoid hemorrhage,SAH)后早期脑损伤(early brain injury,EBI)中脑水肿、运动功能和水通道蛋白4(AQP-4)的影响。方法:108只C57BL/6J小鼠分成假手术对照组(sham组)36只、生理盐水组(SAH+生理盐水组)36只、治疗组(SAH+COG1410组)36只,分别采用血管内穿刺法建立SAH模型。36只小鼠采用类似血管内穿刺法但不刺破颈内动脉建成假手术对照组(sham组);36只小鼠采用血管内穿刺法建成SAH模型,并在1d、2d、3d时经尾静脉向小鼠体内注入生理盐水建成生理盐水组(SAH+生理盐水组);36只小鼠采用血管内穿刺法建成SAH模型在1d、2d、3d时经尾静脉向小鼠体内注入相同剂量的载脂蛋白E短肽COG1410建成治疗组(SAH+COG1410组)。随机选取6只假手术对照组小鼠、6只生理盐水组小鼠、6只治疗组小鼠用于观察小鼠1d、2d、3d的运动功能变化。随机选取6只假手术对照组小鼠、6只生理盐水组小鼠、6只治疗组小鼠采用干湿质量法测定小鼠1d、2d、3d的脑组织含水量的变化。随机选取6只假手术对照组小鼠、6只生理盐水组小鼠、6只治疗组小鼠采用免疫组化法测定小鼠1d、2d、3d的脑组织表达AQP-4阳性细胞数。随机选取6只假手术对照组小鼠、6只生理盐水组小鼠、6只治疗组小鼠采用酶联免疫吸附法(ELISA)测定小鼠1d、2d、3d的脑组织AQP-4蛋白含量。随机选取6只假手术对照组小鼠、6只生理盐水组小鼠、6只治疗组小鼠采用实时定量PCR测定小鼠1d、2d、3d的脑组织AQP-4 mRNA的变化。结果:SAH后生理盐水组(SAH+生理盐水组)和治疗组(SAH+COG1410组)1d、2d、3d神经功能损伤严重性评分均明显高于假手术对照组(sham组)所对应相应时间点的神经功能损伤严重性评分,且两者之间差异有统计学意义(P0.05);生理盐水组(SAH+生理盐水组)1d、2d、3d神经功能损伤严重性评分均明显高于治疗组(SAH+COG1410组)所对应相应时间点的神经功能损伤严重性评分,且两者之间差异有统计学意义(P0.05);小鼠在SAH后神经功能损伤严重性评分在1d时明显增加,2d时达到高峰,之后逐渐下降。SAH后生理盐水组(SAH+生理盐水组)和治疗组(SAH+COG1410组)1d、2d、3d脑组织含水量均明显高于假手术对照组(sham组)所对应相应时间点的脑组织含水量,且差异有统计学意义(P0.05);生理盐水组(SAH+生理盐水组)1d、2d、3d脑组织含水量均明显高于治疗组(SAH+COG1410组)所对应相应时间点的脑组织含水量,且两者之间差异有统计学意义(P0.05);小鼠在SAH后脑组织含水量在1 d时明显增加,2d时达到高峰,之后逐渐下降。SAH后生理盐水组(SAH+生理盐水组)和治疗组(SAH+COG1410组)1d、2d、3d脑组织AQP4蛋白含量均明显高于假手术对照组(sham组)所对应相应时间点的脑组织AQP4蛋白含量,且差异有统计学意义(P0.05);生理盐水组(SAH+生理盐水组)1d、2d、3d脑组织AQP4蛋白含量均明显高于治疗组(SAH+COG1410组)所对应相应时间点的脑组织AQP4蛋白含量,且两者之间差异有统计学意义(P0.05);小鼠在SAH后脑组织AQP4蛋白含量在1 d时明显增加,2d时达到高峰,之后逐渐下降。SAH后生理盐水组(SAH+生理盐水组)和治疗组(SAH+COG1410组)1d、2d、3d脑组织AQP4阳性细胞数均明显高于假手术对照组(sham组)所对应相应时间点的脑组织AQP4阳性细胞数,且差异有统计学意义(P0.05);生理盐水组(SAH+生理盐水组)1d、2d、3d脑组织AQP4阳性细胞数均明显高于治疗组(SAH+COG1410组)所对应相应时间点的脑组织AQP4阳性细胞数,且两者之间差异有统计学意义(P0.05);小鼠在SAH后脑组织AQP4阳性细胞数在1d时明显增加,2d时达到高峰,之后逐渐下降。SAH后生理盐水组(SAH+生理盐水组)和治疗组(SAH+COG1410组)1d、2d、3d脑组织AQP4mRNA表达量均明显高于假手术对照组(sham组)所对应相应时间点的脑组织AQP4mRNA表达量,且差异有统计学意义(P0.05);生理盐水组(SAH+生理盐水组)1d、2d、3d脑组织AQP4mRNA表达量均明显高于治疗组(SAH+COG1410组)所对应相应时间点的脑组织AQP4mRNA表达量,且两者之间差异有统计学意义(P0.05);小鼠在SAH后脑组织AQP4mRNA表达量在1 d时明显增加,2d时达到高峰,之后逐渐下降。结论:载脂蛋白E短肽COG1410可能通过抑制AQP-4的过表达缓解EBI中的脑水肿。
[Abstract]:Objective: To investigate the effect of apolipoprotein E peptide COG1410 on mice with experimental subarachnoid hemorrhage (subarachnoid hemorrhage, SAH) after early brain injury (early brain, injury, EBI) of brain edema, and motor function of aquaporin 4 (AQP-4) effect. Methods: 108 C57BL/6J mice were divided into sham operation group (sham group 36), saline group (saline group SAH+ = 36), treatment group (group SAH+COG1410) 36, SAH models were established in.36 mice by using similar endovascular puncture but not puncture of internal carotid artery into the sham operation group by endovascular puncture (sham group); 36 mice with vascular in the puncture built SAH model, and in 1D, 2D, 3D into the tail vein of mice injected saline into saline group (SAH+ saline group); 36 mice by endovascular puncture method to build SAH model in 1D, 2D, 3D by intravenous injection to mice. The same dose of apolipoprotein E peptide COG1410 into the treatment group (SAH+COG1410 group). 6 rats were randomly selected in sham control mice, 6 mice in the control group, 6 treatment group were used to observe the changes of mouse 1D, 2D, 3D's motion function. Randomly selected 6 rabbits in sham control mice, 6 only the mice in the control group, 6 mice treated by dry wet weight method for the determination of mouse 1D, 2D, 3D changes the water content of the brain tissue. Randomly selected 6 rabbits in sham control mice, 6 mice in the control group, 6 mice treated with immunohistochemical determination of 1D mice, 2D expression the number of AQP-4 positive cells in 3D brain tissue. Randomly selected 6 rabbits in sham control mice, 6 mice in the control group, 6 mice treated by enzyme linked immunosorbent assay (ELISA) determination of mouse 1D, 2D, 3D of brain tissue AQP-4 protein content. Randomly selected 6 rabbits sham operation control group mice. Only 6 students The mice in the control group and treatment group, 6 mice were determined by real-time quantitative 1D, PCR 2D, 3D AQP-4 mRNA changes in brain tissue. Results: SAH after saline group (saline group SAH+) and treatment group (SAH+COG1410 group) 1D, 2D, 3D nerve function injury severity score were significantly higher than that of the sham operation control group (Group sham) corresponding to the corresponding time points of the nerve function injury severity score, there were statistically significant differences between them (P0.05); saline group (saline group SAH+) 1D, 2D, 3D nerve function injury severity score were significantly higher than the treatment group (group SAH+COG1410) corresponding to at the same time the neurological injury severity score, there were statistically significant differences between them (P0.05); mice in SAH after nerve injury severity score increased significantly at 1D, peaked at 2D, then gradually decreased after.SAH under saline group (saline SAH+ Group) and treatment group (SAH+COG1410 group) 1D, 2D, 3D and brain water content were significantly higher than those in sham control group (Group sham) corresponding to the corresponding time points of the water content of brain tissue, and the difference was statistically significant (P0.05); saline group (saline group SAH+) 1D, 2D, 3D in the brain the tissue water content were significantly higher than the treatment group (group SAH+COG1410) corresponding to the corresponding time points of the water content of brain tissue, there were statistically significant differences between them (P0.05); SAH in mouse brain tissue water content at 1 D was significantly increased, reached the peak at 2D, then gradually decreased after.SAH under saline group (SAH+ the physiological saline group) and treatment group (SAH+COG1410 group) 1D, 2D, AQP4 protein content of 3D in brain tissue were significantly higher than those in sham control group (Group sham) corresponding to the corresponding time points of brain tissue AQP4 protein content, and the difference was statistically significant (P0.05); saline group (saline group 1D, SAH+) 2D, 3D brain AQP4 protein content were significantly higher than the treatment group (group SAH+COG1410) corresponding to the corresponding time points of brain tissue AQP4 protein content, there were statistically significant differences between them (P0.05); the content of AQP4 protein in SAH mouse brain tissue at 1 D significantly increased, reached the peak at 2D, then gradually decreased in saline group after.SAH (SAH+ saline group) and treatment group (SAH+COG1410 group) 1D, 2D, AQP4 the number of 3D positive cells in brain tissue were significantly higher than those in sham control group (Group sham) corresponding to the number of AQP4 positive cells in brain tissue at different time points, and the difference was statistically significant (P0.05); saline group (saline SAH+ group 1D, 2D, AQP4) the number of 3D positive cells in brain tissue were significantly higher than the treatment group (group SAH+COG1410) corresponding to the number of AQP4 positive cells in brain tissue at the same time, there were statistically significant differences between them (P0.05); SAH in mice brain tissue AQP4 positive The number of cells in 1D was significantly increased, reached the peak at 2D, then decreased gradually after.SAH saline group (SAH+ saline group) and treatment group (SAH+COG1410 group) 1D, 2D, 3D in brain tissue of AQP4mRNA expression was significantly higher than that in the sham operation group (Group sham) corresponding to the corresponding time points of the brain tissue AQP4mRNA the expression, and the difference was statistically significant (P0.05); saline group (saline group SAH+) 1D, 2D, 3D in brain tissue of AQP4mRNA expression was significantly higher than the treatment group (group SAH+COG1410) corresponding to the corresponding time points of the expression of AQP4mRNA in brain tissue, there were statistically significant differences between them (P0.05); mice the expression of SAH in brain tissue AQP4mRNA content at 1 D was significantly increased, reached the peak at 2D, then gradually decreased. Conclusion: apolipoprotein E peptide COG1410 EBI may alleviate cerebral edema by inhibiting the expression of AQP-4.

【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R743.35

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