去骨瓣减压联合亚低温治疗对重型颅脑损伤大鼠TRPM2、caspase-9表达的影响

发布时间：2018-06-29 15:32

本文选题：重型颅脑损伤 + 去骨瓣减压　；参考：《川北医学院》2017年硕士论文

【摘要】：目的:采用改良的Feeney's自由落体损伤装置建立重型颅脑损伤动物模型,运用此模型进行实验研究:1.亚低温对重型颅脑损伤后大鼠TRPM2、caspase-9表达的影响;2.去骨瓣减压术对重型颅脑损伤后大鼠TRPM2、caspase-9表达的影响;3.亚低温治疗联合去骨瓣减压术对重型颅脑损伤后大鼠TRPM2、caspase-9表达的影响;4.通过TRPM2、caspase-9的表达比较单纯两种治疗及联合治疗方式对重型颅脑损伤疗效的影响;5.为TRPM2通道靶点在STBI治疗上的深入研究提供依据。方法:选取SPF(specific pathogen free)(清洁)级成年SD(Sprague-Dawley)雄性大鼠192只,体重280g~340g。随机分为4组,颅脑损伤组(STBI组即仅钻孔打击)48只,颅脑损伤后亚低温治疗组(STBI+MHT组)48只,颅脑损伤后去骨瓣减压治疗组(STBI+DC组)48只,颅脑损伤后亚低温联合去骨瓣减压组(STBI+MHT+DC组)48只,各组又根据打击后不同时间点分为6个亚组,分别为打击后6h、12h、24h、48h、72h和168h,每个时间点8只大鼠。损伤后不同时间点进行断头,取脑损伤处脑组织来测定脑组织含水量(干湿称重法)、检测TRPM2、casepase-9的表达(免疫组化法)。结果:1.干湿称重法结果:STBI组脑组织含水量:伤后6h开始增加,12h进一步增加,24h达高峰,48h开始降低,72h明显降低,168h降低更明显。STBI+MHT组伤后6h、12 h递增,24h达到高峰,48h、72h递减,168h仍有水肿,与STBI组比较,各时间点脑组织含水量有显著差异(p0.05);STBI+DC组伤后6h、12h递增,24h达到高峰,48h、72h递减,168h仍有水肿,与TBI组比较,各时间点脑组织含水量有显著差异(p0.05);与STBI+MHT组比较,各时间点脑组织含水量有显著差异(p0.05);STBI+MHT+DC组伤后6h、12h递增,24h达到高峰,48h、72h递减,168h趋于正常,与STBI组比较各时间点脑组织含水量有显著差异(p0.05);与STBI+MHT组及TBI+DC组比较,伤后各时间点脑组织含水量有显著差异(p0.05)。2.免疫组化法结果:TRPM2的阳性细胞表达随时间的变化而变化,在伤后6h、12h、24h、48h、72h、168h的各时间点,TRPM2的表达呈现出先升高后降低的趋势,伤后6h即有升高,24h达到高峰,伤后168h降至最低。STBI+MHT组与STBI组比较各时间点TRPM2表达有显著差异(p0.05);STBI+DC组与STBI组比较各时间点TRPM2表达有显著差异(p0.05);STBI+DC组与STBI+MHT组比较各时间点TRPM2表达有显著差异(p0.05);STBI+MHT+DC组与STBI组比较各时间点TRPM2表达有显著差异(p0.05);STBI+MHT+DC组与STBI+DC组各时间点TRPM2表达有显著差异(p0.05);STBI+MHT+DC组与STBI+MHT组各时间点TRPM2表达有显著差异(p0.05)。Casepase-9的阳性细胞表达随时间的变化而变化,在伤后6h、12 h、24h、48h、72h、168h的各时间点,casepase-9的表达呈现出先升高后降低的趋势,伤后6h即有升高,24h达到高峰,伤后168h降至最低。STBI+MHT组与STBI组比较各时间点casepase-9表达有显著差异(p0.05);STBI+DC组与STBI组比较各时间点casepase-9表达有显著差异(p0.05);STBI+DC组与STBI+MHT组比较各时间点有显著差异(p0.05);STBI+MHT+DC组与STBI组比较各时间点casepase-9表达有显著差异(p0.05);STBI+MHT+DC组与STBI+DC组各时间点casepase-9表达有显著差异(p0.05);STBI+MHT+DC组与STBI+MHT组各时间点casepase-9表达有显著差异(p0.05)。结论:1.本实验方法采用改良Feeney's法自由落体制作颅脑损伤模型,成功建立大鼠重型颅脑损伤模型,其制作方法具有简单、可控性好、可重复性好的特点;2.颅脑损伤后氧化应激可使过度的TRPM2表达,Caspase-9表达水平相应增加,促使细胞凋亡,加重继发性脑损伤;3.去大骨瓣减压、亚低温治疗均对重型颅脑损伤具有治疗作用,且两种治疗方法具有协同作用,联合治疗优于单一治疗。
[Abstract]:Objective: to establish an animal model of severe craniocerebral injury with a modified Feeney's free falling body damage device and use this model to study the effect of 1. mild hypothermia on the expression of TRPM2 and caspase-9 in rats after severe craniocerebral injury; (2.) the effect of bone flap decompression on the expression of TRPM2 and caspase-9 in rats after severe craniocerebral injury; 3. hypothermia therapy The effect of combined bone flap decompression on the expression of TRPM2 and caspase-9 in rats after severe craniocerebral injury; 4. through the expression of TRPM2 and caspase-9, the effect of pure two treatment and combined therapy on severe craniocerebral injury was compared; 5. the TRPM2 channel targets were provided for in-depth study on STBI treatment. Methods: SPF (specific pathogen FR) was selected. EE) (EE) (clean) grade adult SD (Sprague-Dawley) male rats, body weight 280g~340g. randomly divided into 4 groups, craniocerebral injury group (group STBI or only drilling) 48, 48 after craniocerebral injury, mild hypothermia treatment group (group STBI+MHT), 48 after craniocerebral trauma decompression treatment group (group STBI+DC), mild hypothermia combined with bone flap decompression group after craniocerebral injury (STB) Group I+MHT+DC) 48, each group was divided into 6 subgroups according to the different time points after the attack, which were after the attack of 6h, 12h, 24h, 48h, 72h and 168h, and 8 rats at each time point. After the injury, the brain tissue was taken to determine the water content of the brain tissue (dry wet weighing method), and the expression of TRPM2 and casepase-9 (immunohistochemistry) was detected. Results: 1. the results of dry and wet weighing method: the water content of brain tissue in group STBI: after injury, 6h began to increase, 12h increased further, 24h reached its peak, 48h began to decrease, 72h decreased obviously, 168h decreased more obviously after.STBI+MHT group, and the 24h reached the peak, 48H, decreased, and still had edema. Compared with the group, the water content of brain tissue at each time point was significantly worse. In group STBI+DC, 6h, 12h increased, 24h reached the peak, 48h, 72h decreased, 168h still had edema. Compared with the TBI group, the water content of the brain tissue was significantly different at each time point (P0.05). Compared with the STBI+MHT group, the water content of the brain tissue was significantly different (P0.05) at each time point (P0.05). Compared with group STBI and group TBI+DC, the water content of brain tissue was significantly different from that of group STBI+MHT and TBI+DC (P0.05).2. immunohistochemical method: the expression of positive cells in TRPM2 changes with time, and 6h, 12h, 24h, 48h, 72h, and all time points after injury. The expression of TRPM2 showed a tendency to increase first and then decrease, and the 6h increased after injury, and the 24h reached the peak. The 168h decreased to the lowest.STBI+MHT group after the injury and the TRPM2 expression at each time point in the.STBI+MHT group was significantly different (P0.05). The STBI+DC group compared with the STBI group, there was a significant difference (P0.05) in each time point (P0.05). The expression of TRPM2 was significantly different (P0.05), and there was significant difference in TRPM2 expression between group STBI+MHT+DC and STBI group at each time point (P0.05); the expression of TRPM2 in STBI+MHT+DC group and STBI+DC group at each time point was significantly different (P0.05), and the expression of positive cells in STBI+MHT+DC group and STBI+MHT group The changes in the time points of 6h, 12 h, 24h, 48h, 72h, 168h after injury, the expression of casepase-9 showed a tendency to increase first and then decrease, and 6h increased after injury, 24h reached its peak, and 168h decreased to the lowest level in.STBI+MHT group after injury. There was significant difference in point casepase-9 expression (P0.05), and there was significant difference in time points between group STBI+DC and STBI+MHT group (P0.05), and there was significant difference in casepase-9 expression between STBI+MHT+DC and STBI groups at each time point (P0.05), and there was a significant difference between STBI+MHT+DC and STBI+DC groups at each time point. The expression of time point casepase-9 was significantly different (P0.05). Conclusion: 1. the experimental method used the modified Feeney's free falling body to make the brain injury model, and successfully established the model of severe brain injury in rats. The production method was simple, good controllability and reproducible characteristics. 2. the oxidative stress after craniocerebral injury can make the excessive expression of TRPM2, Ca The expression level of spase-9 increases correspondingly, promotes cell apoptosis and aggravates secondary brain injury; 3. decompression of large bone flap and mild hypothermia therapy have therapeutic effect on severe craniocerebral injury, and two kinds of treatment methods have synergistic effect, combined therapy is superior to single treatment.
【学位授予单位】：川北医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R651.15

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