本文選題：腦缺血/再灌注　切入點：電針預處理　出處：《大連醫(yī)科大學》2014年碩士論文　論文類型：學位論文

【摘要】：目的：探討電針預處理對腦缺血/再灌注小鼠海馬及皮質區(qū)神經元單磷酸腺苷激活蛋白激酶(AMP-activated protein kinase，AMPK)、神經行為學評分、神經元凋亡及其凋亡蛋白Bax、caspase-9、caspase-3表達的影響。 方法：健康雄性C57BL6小鼠80只，7周齡，采用隨機數字表法，將其隨機分為5組：空白對照(N)組：對小鼠不施加任何處理措施；假手術(S)組：僅分離雙側頸總動脈作對照；腦缺血/再灌注(I/R)組：建立腦缺血模型前無電針預處理；電針百會穴(EA+I/R)組：建立模型前行電針百會穴預處理；電針無關穴位(SEA+I/R)組：建立模型前針刺百會穴旁2mm處以作對照。針刺方式：選取“百會”穴，，在建立腦缺血/再灌注模型前使用0.25mm×40mm的毫針，于頭頂部兩耳連線中點沿皮平刺入2mm，選取疏密波，強度1mA，頻率2/15Hz，以小鼠胡須輕微抖動作為針刺有效的標志，每天1次，每次持續(xù)30min，持續(xù)5天，待最后一次電針刺激結束后24小時，建立有效的前腦缺血模型。對SEA+I/R組小鼠，針刺百會穴旁2mm，其他處理與EA+I/R組相同。采用雙側頸總動脈阻塞(bilateral common carotid artery occlusion，BCCAO)法建立前腦缺血模型，腦缺血15min再灌注。觀察指標：(1)神經行為學評分：3天后對所有小鼠行神經行為學評分；(2)海馬及皮質區(qū)神經元形態(tài)、凋亡神經元計數：對小鼠行神經行為學評分后，每組隨機取8只行甲醛固定后制作石蠟切片，HE染色觀察海馬及皮質神經元形態(tài)，TUNEL法檢測凋亡神經元；(3)海馬及皮質區(qū)總AMPK、pAMPK、Bax、caspase-9、caspase-3蛋白的表達：每組其余8只提取新鮮海馬及皮質腦組織，PCR法測AMPK2mRNA的表達，western blot法測總AMPK、pAMPK、Bax、caspase-9、caspase-3蛋白的表達。 結果：(1)與N組和S組比較，I/R組、EA+I/R組、SEA+I/R組的神經行為學評分顯著升高(P0.05)，海馬、皮質區(qū)凋亡細胞和Bax、caspase-9、caspase-3表達也升高(P0.05)， pAMPK蛋白水平升高(P0.05)。 (2)與I/R組、SEA+I/R組相比，EA+I/R組神經行為學評分降低(P0.05)，海馬、皮質區(qū)凋亡細胞和Bax、caspase-9、caspase-3表達降低(P0.05)，pAMPK蛋白水平升高(P0.05)。 (3)各組間AMPK2mRNA及總AMPK蛋白的表達差異無統計學意義(P0.05)。 結論：電針預處理能激活腦缺血/再灌注小鼠海馬及皮質區(qū)AMPK，升高pAMPK水平，發(fā)揮腦保護作用。 目的：探討針刺對老年髖關節(jié)置換術患者血清神經元特異性烯醇化酶(NSE)、S-100β蛋白濃度與術后認知功能障礙之間的關系。 方法：一般資料：86例擇期腰麻-硬膜外聯合阻滯麻醉下行髖關節(jié)年患者，年齡70-80歲，體重48-89公斤，ASA分級Ⅱ～Ⅲ級，手術中鎮(zhèn)痛完善，咪達唑侖鎮(zhèn)靜。隨機分為2組：對照組(N組)46例，觀察組(針刺組A組)40例。觀察項目：(1)運用簡易智能量表(mini-mental state examination, MMSE)對患者于術前1天(T0)，術后3天(T1)及術后7天(T2)進行認知功能評分。(2) POCD：比較兩組術后3天(T1)及術后7天(T2)POCD的發(fā)生率；(3) NSE、S-100β蛋白的濃度：兩組患者麻醉前(t0)、術后6小時(t1)、術后24小時(t3)及48小時(t4)分別抽取靜脈血，監(jiān)測NSE、S-100β血清濃度；(4)比較兩組手術時間，術中咪達唑侖用量、輸液量、出血量、尿量、阿托品及麻黃堿使用率及其他嚴重并發(fā)癥。 結果：(1) POCD：與N組相比，A組在術后T1及T2時POCD發(fā)生率明顯降低(P0.05)；(2) NSE、S-100β蛋白的濃度：與N組相比，A組血清NSE濃度在t2-t4時降低(P0.05)，S-100β蛋白的濃度在t1-t4時降低(P0.05)；組內比較，N組NSE在t2-t4時較t0時升高， S-100β蛋白濃度在t1-t4時較t0時升高。A組血清NSE濃度在t2、t3時較t0時升高(P0.05)，S-100β蛋白濃度在t1、t4時較t0時升高(P0.05)，(3)兩組手術時間、術中咪達唑侖用量、輸液量、出血量、尿量、阿托品及麻黃堿使用率差異無統計學意義(P0.05)；并且兩組均沒有嚴重的并發(fā)癥。 結論：針刺能夠降低老年髖關節(jié)置換術患者血清神經元特異性烯醇化酶(NSE)、S-100β蛋白，改善術后認知功能，有利于患者康復。
[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.
鏂規(guī)硶錛氫竴鑸祫鏂欙細86渚嬫嫨鏈熻叞楹

本文編號：1642374