不同等级闭合性脑损伤大鼠HSP70的表达变化及意义

发布时间：2018-05-24 12:26

本文选题：HSP70 + 闭合性脑损伤　；参考：《南昌大学》2010年硕士论文

【摘要】： 目的:探索HSP70蛋白及其mRNA在大鼠不同等级闭合性脑损伤中的表达;探索大鼠闭合性脑损伤分级自由落体打击技术;探讨HSP70在大鼠不同等级闭合性脑损伤中的意义。方法:1、根据自由落体原理,采用不同重量的砝码从同一高度下落,以不同冲击应力(128.90Kpa、386.70Kpa、644.50Kpa)的方法,造成轻、中、重度大鼠闭合性脑损伤模型共54只为实验组,颅脑假手术18只大鼠为对照组。2、应用免疫组织化学EliVision方法,检测不同等级大鼠闭合性脑损伤中HSP70蛋白的表达变化情况。3、采用逆转录多聚酶链反应(RT-PCR)技术对不同等级大鼠闭合性脑损伤中HSP70mRNA进行定量分析。结果:1、HE染色结果显示伤后3h轻度损伤组软脑膜下皮质出血,神经胶质细胞增生,中度损伤组皮质浅层出血,脑实质水肿,神经胶质细胞增生,重度损伤组皮质广泛出血,出血灶周围见中性粒细胞浸润,神经元消失,周边脑实质网格样。2、免疫组织化学染色结果显示伤后3h HSP70蛋白在对照组、轻度损伤组、中度损伤组及重度损伤组均有不同程度的表达,其阳性表达率分别为8.1%、11.24%、20.56%、30.43%;轻度损伤组与对照组之间表达差异无统计学意义(P0.05);中度、重度损伤组与对照组表达差异有统计学意义(P0.05);轻、中、重度损伤组之间表达差异有统计学意义(P0.05)。3、RT-PCR结果显示伤后3 h HSP70mRNA在对照组、轻度损伤组、中度损伤组及重度损伤组均有不同程度的表达,HSP70与β-actin的光密度比值平均值分别为0.330、0.415、0.550、0.767;轻度损伤组与对照组之间比值差异无统计学意义(P0.05);中度、重度损伤组与对照组比值差异有统计学意义(P0.05);轻、中、重度损伤组之间比值差异有统计学意义(P0.05)。结论: 1、HSP70mRNA及蛋白在应激状态下的神经元损伤的表达是一致的,表明短时间内HSP70mRNA及其蛋白表达率可随着损伤程度的加重明显升高,HSP70的表达差异可作为短时间内中度以上闭合性脑损伤程度的一个较好的分级指标。2、该模型解决并完善了经典落体打击法中打击部位不稳定的问题,重复性好,对闭合性脑损伤实验研究有实用价值。
[Abstract]:Objective: to investigate the expression of HSP70 protein and its mRNA in rat brain injury of different grades, to explore the technique of free falling body attack in different grades of closed brain injury in rats, and to explore the significance of HSP70 in different grades of closed brain injury in rats. Methods according to the principle of free falling body, 54 rats with light, moderate and severe closed brain injury were induced by using different weights falling from the same height and different impact stress (128.90Kpa-386.70Kpa-644.50Kpaa). Eighteen rats with craniocerebral sham-operation were used as control group (n = 18). Immunohistochemical EliVision method was used. The changes of HSP70 protein expression in closed brain injury of rats of different grades were detected. Reverse transcriptase polymerase chain reaction (RT PCR) technique was used to quantitatively analyze HSP70mRNA in closed brain injury of rats of different grades. Results the results showed that the subpial cortex hemorrhage, glial cell proliferation, superficial cortical hemorrhage, cerebral parenchyma edema, glial cell proliferation and extensive hemorrhage in severe injury group were found in mild injury group 3 hours after injury. Neutrophil infiltration and neuronal disappearance were observed around the hemorrhage foci. The results of immunohistochemical staining showed that HSP70 protein was present in the control group and mild injury group at 3 h after injury. The positive expression rates of moderate injury group and severe injury group were 8.1and 11.240.56and 30.433.There was no significant difference between mild injury group and control group (P 0.05). There was significant difference in the expression of HSP70mRNA between the severe injury group and the control group (P 0.05), and between the mild, moderate and severe injury groups, the RT-PCR results showed that the expression of HSP70mRNA was in the control group and the mild injury group at 3 hours after injury. The average optical density ratio of HSP70 and 尾 -actin in moderate injury group and severe injury group were 0.330, 0.415, 0.550 and 0.767.There was no significant difference between mild injury group and control group (P 0.05). There was significant difference in the ratio between severe injury group and control group (P 0.05), while there was significant difference between light, moderate and severe injury group (P 0.05). Conclusion: 1 the expression of HSP70 mRNA and protein in the injured neurons under stress is consistent. The results indicate that the expression rate of HSP70mRNA and its protein can increase significantly with the severity of injury in a short period of time, and the difference of HSP70 expression can be used as a good grading index of the degree of closed brain injury in a short period of time. Improved the problem of unstable hit parts in the classical falling body attack method, It has good reproducibility and has practical value for the experimental study of closed brain injury.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R363

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