tPA介导的BDNF转换障碍在异丙酚致大鼠脑功能障碍中的机制研究
发布时间：2018-11-12 06:50
【摘要】：目的探讨异丙酚对新生大鼠海马tPA表达的影响以及是否通过影响tPA表达、阻碍脑发育高峰期海马proBDNF向mBDNF的成熟转换,最终导致大鼠学习记忆的受损。方法SD大鼠162只,年龄7天,SPF级,体重介于12g到16g之间,雌性雄性参半。随机将其分为三组(每组n=54只):生理盐水组(C组)为连续7天注射0.9%NaCl溶液。单次异丙酚注射组(SP组)为前6天连续注射0.9%NaCl溶液,第7天注射异丙酚。多次异丙酚注射组(RP组)为连续7天注射异丙酚。各组大鼠均采用腹腔注射异丙酚75 mg/kg或0.9%NaCl 7.5 ml/kg。随机地选取每组六只大鼠行血糖和血气测定。建模完成的17d行水迷宫测试大鼠的学习与记忆能力。HE染色法以及尼氏染色法观测海马的形态结构。Western blolt法检测建模完成的2 h、24 h、48 h、72 h、22 d五个时间点下tPA蛋白表达的动态变化,RT-PCR法测定tPA mRNA的改变。Western blolt法检测建模完成后22 d时pro BDNF、mBDNF、Caspase3蛋白表达改变。TUNEL法测定神经元的凋亡。结果与C组和SP组比较,RP组逃逸时间增加,且在原平台象限的探索时间以及穿越原平台所在位置的次数减少(P0.05);RP组tPA的protein表达于五个时间点均呈显著地下降趋势;tPA的mRNA表达也出现明显的下调(P0.05);伴随tPA、mBDNF表达的减弱,proBDNF表达显著增强(P0.05);RP组海马神经细胞数量减少且排列紊乱,神经元内尼氏体明显减少,部分神经元出现变性及坏死;神经元凋亡细胞数量增多,pro-Caspase3表达减低,cleaved-Caspase3表达增高(P0.05)。而SP组与C组比较仅24 h时tPA的protein表达呈下降趋势(P0.05),其他各结果比较均没有统计学显著性(P0.05)。结论多次注射异丙酚后导致新生大鼠远期空间学习记忆能力的下降,其中的机制可能与海马tPA表达下调、海马pro BDNF向mBDNF成熟转换障碍以及海马神经元正常形态及功能破坏、神经元凋亡发生有关;而单次注射异丙酚对此影响不显著。
[Abstract]:Objective to investigate the effect of propofol on the expression of tPA in hippocampus of neonatal rats and whether the effect of propofol on the expression of tPA hinders the maturation of proBDNF to mBDNF in hippocampus during the peak period of brain development, resulting in the impairment of learning and memory in rats. Methods 162 SD rats, aged 7 days, with SPF grade, weighing between 12 g and 16 g, were divided into two groups: female and male. They were randomly divided into three groups (n = 54 in each group): saline group (C group) was injected with 0.9%NaCl solution for 7 days. The single propofol injection group (SP group) was continuously injected with 0.9%NaCl solution on the first 6 days, and propofol was injected on the 7th day. Multiple propofol injection group (RP group) was injected with propofol for 7 days. Rats in each group were given intraperitoneal injection of propofol for 75 mg/kg or 0.9%NaCl 7.5 ml/kg.. Six rats in each group were randomly selected to measure blood glucose and blood gas. The learning and memory abilities of the rats were tested by water maze on 17 days after modeling. The morphologic structure of hippocampus was observed by HE staining method and Nissl staining method. The morphologic structure of hippocampus was detected by. Western blolt method at 2 h, 24 h, 48 h and 72 h, respectively. The dynamic changes of tPA protein expression at five time points at 22 d, the change of tPA mRNA by RT-PCR method, the expression of pro BDNF,mBDNF,Caspase3 protein at 22 d after modeling were detected by. Western blolt method, and the apoptosis of neurons by TUNEL method. Results compared with group C and group SP, the escape time of RP group increased, and the exploring time in the original platform quadrant and the times of crossing the original platform location decreased (P0.05) the protein expression of tPA in); RP group was significantly decreased at five time points. The expression of mRNA in tPA was also down-regulated (P0.05), and the expression of proBDNF was significantly increased with the decrease of tPA,mBDNF expression (P0.05). In RP group, the number of hippocampal neurons decreased and the number of neurons was disordered, and the number of neuronal apoptotic cells increased, the expression of pro-Caspase3 decreased and the expression of cleaved-Caspase3 increased (P0.05). Compared with C group, the protein expression of tPA in SP group decreased at 24 h (P0.05), and there was no significant difference in other results (P0.05). Conclusion after multiple injections of propofol, the long-term spatial learning and memory ability of neonatal rats is decreased, which may be related to the down-regulation of tPA expression in hippocampus, the disturbance of maturation of pro BDNF to mBDNF in hippocampus and the normal morphology and function of hippocampal neurons. Neuronal apoptosis is related to neuronal apoptosis. But single injection of propofol had no significant effect on it.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R614
本文编号：2326362
[Abstract]:Objective to investigate the effect of propofol on the expression of tPA in hippocampus of neonatal rats and whether the effect of propofol on the expression of tPA hinders the maturation of proBDNF to mBDNF in hippocampus during the peak period of brain development, resulting in the impairment of learning and memory in rats. Methods 162 SD rats, aged 7 days, with SPF grade, weighing between 12 g and 16 g, were divided into two groups: female and male. They were randomly divided into three groups (n = 54 in each group): saline group (C group) was injected with 0.9%NaCl solution for 7 days. The single propofol injection group (SP group) was continuously injected with 0.9%NaCl solution on the first 6 days, and propofol was injected on the 7th day. Multiple propofol injection group (RP group) was injected with propofol for 7 days. Rats in each group were given intraperitoneal injection of propofol for 75 mg/kg or 0.9%NaCl 7.5 ml/kg.. Six rats in each group were randomly selected to measure blood glucose and blood gas. The learning and memory abilities of the rats were tested by water maze on 17 days after modeling. The morphologic structure of hippocampus was observed by HE staining method and Nissl staining method. The morphologic structure of hippocampus was detected by. Western blolt method at 2 h, 24 h, 48 h and 72 h, respectively. The dynamic changes of tPA protein expression at five time points at 22 d, the change of tPA mRNA by RT-PCR method, the expression of pro BDNF,mBDNF,Caspase3 protein at 22 d after modeling were detected by. Western blolt method, and the apoptosis of neurons by TUNEL method. Results compared with group C and group SP, the escape time of RP group increased, and the exploring time in the original platform quadrant and the times of crossing the original platform location decreased (P0.05) the protein expression of tPA in); RP group was significantly decreased at five time points. The expression of mRNA in tPA was also down-regulated (P0.05), and the expression of proBDNF was significantly increased with the decrease of tPA,mBDNF expression (P0.05). In RP group, the number of hippocampal neurons decreased and the number of neurons was disordered, and the number of neuronal apoptotic cells increased, the expression of pro-Caspase3 decreased and the expression of cleaved-Caspase3 increased (P0.05). Compared with C group, the protein expression of tPA in SP group decreased at 24 h (P0.05), and there was no significant difference in other results (P0.05). Conclusion after multiple injections of propofol, the long-term spatial learning and memory ability of neonatal rats is decreased, which may be related to the down-regulation of tPA expression in hippocampus, the disturbance of maturation of pro BDNF to mBDNF in hippocampus and the normal morphology and function of hippocampal neurons. Neuronal apoptosis is related to neuronal apoptosis. But single injection of propofol had no significant effect on it.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R614
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