针刺脑保护作用的机制探讨和临床应用
发布时间：2018-03-21 05:04
本文选题：脑缺血/再灌注 切入点：电针预处理 出处：《大连医科大学》2014年硕士论文 论文类型：学位论文
【摘要】：目的：探讨电针预处理对脑缺血/再灌注小鼠海马及皮质区神经元单磷酸腺苷激活蛋白激酶(AMP-activated protein kinase，AMPK)、神经行为学评分、神经元凋亡及其凋亡蛋白Bax、caspase-9、caspase-3表达的影响。 方法：健康雄性C57BL6小鼠80只，7周龄，采用随机数字表法，将其随机分为5组：空白对照(N)组：对小鼠不施加任何处理措施；假手术(S)组：仅分离双侧颈总动脉作对照；脑缺血/再灌注(I/R)组：建立脑缺血模型前无电针预处理；电针百会穴(EA+I/R)组：建立模型前行电针百会穴预处理；电针无关穴位(SEA+I/R)组：建立模型前针刺百会穴旁2mm处以作对照。针刺方式：选取“百会”穴，，在建立脑缺血/再灌注模型前使用0.25mm×40mm的毫针，于头顶部两耳连线中点沿皮平刺入2mm，选取疏密波，强度1mA，频率2/15Hz，以小鼠胡须轻微抖动作为针刺有效的标志，每天1次，每次持续30min，持续5天，待最后一次电针刺激结束后24小时，建立有效的前脑缺血模型。对SEA+I/R组小鼠，针刺百会穴旁2mm，其他处理与EA+I/R组相同。采用双侧颈总动脉阻塞(bilateral common carotid artery occlusion，BCCAO)法建立前脑缺血模型，脑缺血15min再灌注。观察指标：(1)神经行为学评分：3天后对所有小鼠行神经行为学评分；(2)海马及皮质区神经元形态、凋亡神经元计数：对小鼠行神经行为学评分后，每组随机取8只行甲醛固定后制作石蜡切片，HE染色观察海马及皮质神经元形态，TUNEL法检测凋亡神经元；(3)海马及皮质区总AMPK、pAMPK、Bax、caspase-9、caspase-3蛋白的表达：每组其余8只提取新鲜海马及皮质脑组织，PCR法测AMPK2mRNA的表达，western blot法测总AMPK、pAMPK、Bax、caspase-9、caspase-3蛋白的表达。 结果：(1)与N组和S组比较，I/R组、EA+I/R组、SEA+I/R组的神经行为学评分显著升高(P0.05)，海马、皮质区凋亡细胞和Bax、caspase-9、caspase-3表达也升高(P0.05)， pAMPK蛋白水平升高(P0.05)。 (2)与I/R组、SEA+I/R组相比，EA+I/R组神经行为学评分降低(P0.05)，海马、皮质区凋亡细胞和Bax、caspase-9、caspase-3表达降低(P0.05)，pAMPK蛋白水平升高(P0.05)。 (3)各组间AMPK2mRNA及总AMPK蛋白的表达差异无统计学意义(P0.05)。 结论：电针预处理能激活脑缺血/再灌注小鼠海马及皮质区AMPK，升高pAMPK水平，发挥脑保护作用。 目的：探讨针刺对老年髋关节置换术患者血清神经元特异性烯醇化酶(NSE)、S-100β蛋白浓度与术后认知功能障碍之间的关系。 方法：一般资料：86例择期腰麻-硬膜外联合阻滞麻醉下行髋关节年患者，年龄70-80岁，体重48-89公斤，ASA分级Ⅱ～Ⅲ级，手术中镇痛完善，咪达唑仑镇静。随机分为2组：对照组(N组)46例，观察组(针刺组A组)40例。观察项目：(1)运用简易智能量表(mini-mental state examination, MMSE)对患者于术前1天(T0)，术后3天(T1)及术后7天(T2)进行认知功能评分。(2) POCD：比较两组术后3天(T1)及术后7天(T2)POCD的发生率；(3) NSE、S-100β蛋白的浓度：两组患者麻醉前(t0)、术后6小时(t1)、术后24小时(t3)及48小时(t4)分别抽取静脉血，监测NSE、S-100β血清浓度；(4)比较两组手术时间，术中咪达唑仑用量、输液量、出血量、尿量、阿托品及麻黄碱使用率及其他严重并发症。 结果：(1) POCD：与N组相比，A组在术后T1及T2时POCD发生率明显降低(P0.05)；(2) NSE、S-100β蛋白的浓度：与N组相比，A组血清NSE浓度在t2-t4时降低(P0.05)，S-100β蛋白的浓度在t1-t4时降低(P0.05)；组内比较，N组NSE在t2-t4时较t0时升高， S-100β蛋白浓度在t1-t4时较t0时升高。A组血清NSE浓度在t2、t3时较t0时升高(P0.05)，S-100β蛋白浓度在t1、t4时较t0时升高(P0.05)，(3)两组手术时间、术中咪达唑仑用量、输液量、出血量、尿量、阿托品及麻黄碱使用率差异无统计学意义(P0.05)；并且两组均没有严重的并发症。 结论：针刺能够降低老年髋关节置换术患者血清神经元特异性烯醇化酶(NSE)、S-100β蛋白，改善术后认知功能，有利于患者康复。
[Abstract]:Objective: To investigate the effect of electroacupuncture pretreatment on AMP-activated protein kinase (AMPK), neurobehavioral score, neuronal apoptosis and apoptosis protein Bax, caspase-9 and Caspase-3 expression in hippocampus and cortex of mice after cerebral ischemia / reperfusion.
Methods: 80 healthy male C57BL6 mice, aged 7 weeks, were randomly divided into 5 groups: control group (N): without any treatment measures of mice; sham operation group (S): only separated the bilateral common carotid artery as control; cerebral ischemia / reperfusion (I/R) group: establishment of electroacupuncture preconditioning cerebral ischemia; Electroacupuncture Baihui (EA+I/R) group: to establish the model on EA Baihui acupoint electroacupuncture pretreatment; independent (SEA+I/R) group: to establish the model before acupuncture Baihui 2mm by side as control. Methods: selected acupuncture "Baihui" point in the establishment of cerebral ischemia the use of 0.25mm * 40MM / reperfusion model before the needle head at the top two ear piercing 2mm along the midpoint of Pippin, select the density wave, intensity of 1mA, frequency of 2/15Hz, with a slight jitter as mice beard effective acupuncture, 1 times a day, each lasting 30min, lasting 5 days, for the last time Electroacupuncture at 24 hours after the end, the establishment of an effective model of forebrain ischemia. In SEA+I/R group, acupuncture Baihui side 2mm, EA+I/R and other treatments were the same. The bilateral common carotid artery occlusion (bilateral common carotid artery occlusion, BCCAO) forebrain ischemia model was established, 15min in cerebral ischemia reperfusion. Observation index: (1) neurological score: 3 days for all mice underwent neurological behaviors; (2) morphology of neurons in hippocampus and cortex, the number of apoptotic neurons of mouse nerve behavioral score after each group were randomly selected for 8 formaldehyde fixed paraffin sections, observe the neurons of hippocampus and cortex morphology HE staining, detection the apoptosis of neurons by TUNEL; (3) the total AMPK in hippocampus and cortex of pAMPK, Bax, caspase-9, caspase-3 protein expression: the remaining 8 rats in each group were extracted from fresh hippocampus and cortex of brain tissue, the expression of AMPK2mRNA measured by PCR, we The expression of total AMPK, pAMPK, Bax, caspase-9, caspase-3 protein was measured by stern blot.
Results: (1) compared with group N and group S, the neurobehavioral score of group I/R, group EA+I/R and SEA+I/R increased significantly (P0.05), and the expression of apoptotic cells and Bax, caspase-9 and caspase-3 increased in hippocampus (P0.05), and the level of pAMPK protein increased (P0.05).
(2) compared with group I/R and group SEA+I/R, the neurobehavioral score of group EA+I/R decreased (P0.05), and the expression of apoptotic cells and Bax, caspase-9 and caspase-3 decreased (P0.05), and pAMPK protein level increased (P0.05) in hippocampus and cortex.
(3) there was no significant difference in the expression of AMPK2mRNA and total AMPK protein in each group (P0.05).
Conclusion: electroacupuncture preconditioning can activate the AMPK in the hippocampus and cortex of cerebral ischemia / reperfusion mice, increase the level of pAMPK and play the role of brain protection.
Objective: To explore the relationship between serum neuron specific enolase (NSE), S-100 beta protein and postoperative cognitive dysfunction in elderly patients undergoing hip replacement.
鏂规硶锛氫竴鑸祫鏂欙細86渚嬫嫨鏈熻叞楹
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