U50488H对体外循环术后大鼠认知功能与海马细胞凋亡影响
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摘要
目的探讨κ-阿片受体激动剂U50488H对体外循环术后大鼠认知功能与海马细胞凋亡的影响。方法选取清洁级成年雄性SD大鼠48只,随机分为假手术组(S组)、体外循环组(C组)、U50488H+体外循环组(K组)及U50488H+特异性U50488H拮抗剂(Nor-BNI)+体外循环组(NK组),每组各12只。4组大鼠于麻醉置管/体外循环后1 d行水迷宫检测。酶联免疫吸附法检测血清中白细胞介素-6、中枢神经特异蛋白、肿瘤坏死因子α的浓度;免疫蛋白印记检测海马caspase-3表达的变化。结果 C组、K组、NK组的海马神经元凋亡阳性细胞积分吸光度值、caspase-3蛋白表达及血清白细胞介素-6、中枢神经特异蛋白、肿瘤坏死因子α浓度均高于S组; C组、NK组的海马神经元凋亡阳性细胞积分吸光度值、caspase-3蛋白表达及血清白细胞介素-6、中枢神经特异蛋白、肿瘤坏死因子α浓度均高于K组,组间比较,差异均有统计学意义(P <0. 05)。与S组比较,C组、K组、NK组大鼠逃避潜伏期时间明显延长,空间探索试验进入目标象限的次数明显减少;与C组比较,K组大鼠平均逃避潜伏期时间明显缩短,空间探索试验中进入目标象限的次数明显增多,差异有统计学意义(P <0. 05)。结论 U50488H可在一定程度降低体外循环诱发的大鼠海马细胞凋亡及炎症反应,对体外循环后认知功能有一定保护作用。
Objective To investigate the effect of U50488 H on cognitive function and hippocampal cell apoptosis by CPB in rats.Methods Forty-eight healthy adult male SD rats of clean grade were randomly divided into sham operation group( Group S),cardiopulmonary bypass group( Group C),U50488 H + CPB group( Group K),U50488 H + specific U50488 H antagonist + CPB group( Group NK),with 12 rats in each group. The rats in the 4 groups were tested for water maze 1 day after anesthesia catheterization/extracorporeal circulation. Apoptotic morphology of hippocampal tissue was observed by HE staining,IL-6,S100βand TNF-α in serum were detected by ELISA. The expression changes of caspase-3 protein was detected by westen-blot. Results The values of integral-absorbance,expression of caspase-3 protein,IL-6,S100β and TNF-α in hippocampal neurons of Group C,Group K and Group NK were higher than those of Group S. The hippocampal neuron apoptosis positive cell integral absorbance value,caspase-3 protein expression,IL-6,S100β and TNF-α concentration in group C and NK were all higher than those in Group K( P < 0. 05). Compared with Group S,the escape latency time of rats in Group C,Group K and Group NK was significantly prolonged,and the number of space exploration experiments entering the target quadrant was significantly reduced. Compared with Group C,the average escape latency time of rats in Group K was significantly shortened,and the number of entering the target quadrant was significantly increased in the space exploration experiment( P < 0. 05). Conclusion U50488 H could reduce hippocampal cell apoptosis by cardiopulmonary bypass in rats to some extent,and protect the cognitive function in rates.
Objective To investigate the effect of U50488 H on cognitive function and hippocampal cell apoptosis by CPB in rats.Methods Forty-eight healthy adult male SD rats of clean grade were randomly divided into sham operation group( Group S),cardiopulmonary bypass group( Group C),U50488 H + CPB group( Group K),U50488 H + specific U50488 H antagonist + CPB group( Group NK),with 12 rats in each group. The rats in the 4 groups were tested for water maze 1 day after anesthesia catheterization/extracorporeal circulation. Apoptotic morphology of hippocampal tissue was observed by HE staining,IL-6,S100βand TNF-α in serum were detected by ELISA. The expression changes of caspase-3 protein was detected by westen-blot. Results The values of integral-absorbance,expression of caspase-3 protein,IL-6,S100β and TNF-α in hippocampal neurons of Group C,Group K and Group NK were higher than those of Group S. The hippocampal neuron apoptosis positive cell integral absorbance value,caspase-3 protein expression,IL-6,S100β and TNF-α concentration in group C and NK were all higher than those in Group K( P < 0. 05). Compared with Group S,the escape latency time of rats in Group C,Group K and Group NK was significantly prolonged,and the number of space exploration experiments entering the target quadrant was significantly reduced. Compared with Group C,the average escape latency time of rats in Group K was significantly shortened,and the number of entering the target quadrant was significantly increased in the space exploration experiment( P < 0. 05). Conclusion U50488 H could reduce hippocampal cell apoptosis by cardiopulmonary bypass in rats to some extent,and protect the cognitive function in rates.
引文
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[2]Charron C,Messier C,Plamondon H,et al. Neuroprotection and functional recovery conferred by administration of kappa-and deltaopioid agonists in a rat model of global ischemia[J]. Physiol Behav,2015,93(3):502-511.
[3]段铭杰,孙莹杰,郑翔,等.改良穿刺法建立大鼠无血预充非停跳体外循环后认知功能障碍模型[J].中国体外循环杂志,2017,15(3):179-184.
[4]Doble BW,Woodgett JR. GSK-3:tricks of the trade for a multitasking kinase[J]. J Cell Sci,2003,116(7):1175-1186.
[5]Chou CH,Chou AK,Lin CC,et a1. GSK3βregulates BCL2L12 and BCL2L12A anti-alpoposis signaling in glioblastoma and is inhibited by Li CI[J]. Cell Cycle,2012,11(3):532-542.
[6]Undar A,Wang S. Translational research is a necessity for selecting the best components of the extracorporeal circuitry for neonatal and pediatric CPB patients[J]. Perfusion,2013,28(2):171-172.
[7]Duan X,Ji B,Liu G,et al. Evaluation of shunting flow differences in varied conditions in a simulated adult CPB model during normothermia[J]. Perfusion,2011,26(3):207-213.
[8]许德芳.不同全身麻醉药物及针刺干预对体外循环患者围术期氧化应激的影响[D].天津医科大学,2016.
[9]Ramlawi B,Rudolph JL,Mieno S,et al. C-reactive protein and inflammatory response associated to neurocognitive decline following cardiac surgery[J]. Surgery,2016,140(2):221-226.
[10]向玲,孙广运.术后认知功能障碍与麻醉相关性的研究进展[J].中国民族民间医药,2014,23(23):21-23.
[11]Limpachayaporn P,Schfers M,Haufe G. Isatin sulfonamides:potent caspases-3 and-7 inhibitors,and promising PET and SPECT radiotracers for apoptosis imaging[J]. Future Med Chem,2015,7(9):1173-1196.
[12]刘铭,刘欣伟,刘宪民,等. TNF-α、Caspase-3在大鼠脊髓损伤后神经细胞凋亡中相关性研究[J].临床军医杂志,2017,45(4):366-370.
[13]Pastuszko P,Pirzadeh A,Reade E,et al. The effect of hypothermia on neuronal viability following cardiopulmonary bypass and circulatory arrest in newborn piglets[J]. Eur J Cardiothorac Surg,2009,35(4):577-581.