【摘要】：目的探討異丙酚對新生大鼠海馬tPA表達(dá)的影響以及是否通過影響tPA表達(dá)、阻礙腦發(fā)育高峰期海馬proBDNF向mBDNF的成熟轉(zhuǎn)換,最終導(dǎo)致大鼠學(xué)習(xí)記憶的受損。方法SD大鼠162只,年齡7天,SPF級,體重介于12g到16g之間,雌性雄性參半。隨機將其分為三組(每組n=54只):生理鹽水組(C組)為連續(xù)7天注射0.9%NaCl溶液。單次異丙酚注射組(SP組)為前6天連續(xù)注射0.9%NaCl溶液,第7天注射異丙酚。多次異丙酚注射組(RP組)為連續(xù)7天注射異丙酚。各組大鼠均采用腹腔注射異丙酚75 mg/kg或0.9%NaCl 7.5 ml/kg。隨機地選取每組六只大鼠行血糖和血氣測定。建模完成的17d行水迷宮測試大鼠的學(xué)習(xí)與記憶能力。HE染色法以及尼氏染色法觀測海馬的形態(tài)結(jié)構(gòu)。Western blolt法檢測建模完成的2 h、24 h、48 h、72 h、22 d五個時間點下tPA蛋白表達(dá)的動態(tài)變化,RT-PCR法測定tPA mRNA的改變。Western blolt法檢測建模完成后22 d時pro BDNF、mBDNF、Caspase3蛋白表達(dá)改變。TUNEL法測定神經(jīng)元的凋亡。結(jié)果與C組和SP組比較,RP組逃逸時間增加,且在原平臺象限的探索時間以及穿越原平臺所在位置的次數(shù)減少(P0.05);RP組tPA的protein表達(dá)于五個時間點均呈顯著地下降趨勢;tPA的mRNA表達(dá)也出現(xiàn)明顯的下調(diào)(P0.05);伴隨tPA、mBDNF表達(dá)的減弱,proBDNF表達(dá)顯著增強(P0.05);RP組海馬神經(jīng)細(xì)胞數(shù)量減少且排列紊亂,神經(jīng)元內(nèi)尼氏體明顯減少,部分神經(jīng)元出現(xiàn)變性及壞死;神經(jīng)元凋亡細(xì)胞數(shù)量增多,pro-Caspase3表達(dá)減低,cleaved-Caspase3表達(dá)增高(P0.05)。而SP組與C組比較僅24 h時tPA的protein表達(dá)呈下降趨勢(P0.05),其他各結(jié)果比較均沒有統(tǒng)計學(xué)顯著性(P0.05)。結(jié)論多次注射異丙酚后導(dǎo)致新生大鼠遠(yuǎn)期空間學(xué)習(xí)記憶能力的下降,其中的機制可能與海馬tPA表達(dá)下調(diào)、海馬pro BDNF向mBDNF成熟轉(zhuǎn)換障礙以及海馬神經(jīng)元正常形態(tài)及功能破壞、神經(jīng)元凋亡發(fā)生有關(guān);而單次注射異丙酚對此影響不顯著。
[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.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R614

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