“百会”“大椎”穴电针预处理减轻反复丙泊酚麻醉诱导的大鼠学习记忆障碍.pdf

.78“百会”“大椎”穴电针预处理减轻反复丙泊酚麻醉诱导的大鼠学习记忆障碍JCAM.Aug.2023,Vol.39,NO.8范舜钦1,王喜军1，韦松里,陆思施,郭永榕（1.广西中医药大学附属国际壮医医院，广西南宁530 2 0 1;2.广西中医药大学，广西南宁530 2 0 0）摘要目的：观察“百会”“大椎”穴电针预处理减轻反复丙泊酚麻醉诱发大鼠学习记忆障碍的作用。方法：通过给予连续5d丙泊酚麻醉诱导学习记忆障碍模型，在丙泊酚给药前给予电针预处理连续5d完成电针干预。Morris水迷宫检测大鼠学习记忆能力;HE染色用于观察海马神经细胞数量及形态改变；免疫荧光染色检测海马DentateGyrus区磷酸化环磷腺苷效应元件结合蛋白（pCREB）蛋白表达；用Western Blotting测定B细胞淋巴瘤-2（Bcl-2）、半胱氨酸蛋白酶-3（Caspase-3）、B细胞淋巴瘤-2相关X蛋白（Bax）和CREB蛋白的表达情况。结果：Morris水迷宫结果：与对照组比较，丙泊酚组大鼠的穿台次数、目标象限游泳时长及路径占比明显减少，差异具有统计学意义（P0.05）；电针组大鼠的穿台次数、目标象限游泳时长及路径占比均明显高于丙泊酚组，差异具有统计学意义（P0.05）。H E结果：丙泊酚组海马DG区细胞层数减少，细胞密集度降低；电针组海马DG区神经元细胞层数显著恢复，细胞密集度明显升高。免疫荧光结果：与对照组比较，丙泊酚组海马区pCREB蛋白累积荧光强度显著降低，差异具有统计学意义（P0.05）；与丙泊酚组比较，电针组海马区pCREB蛋白累积荧光强度显著升高，差异具有统计学意义(P0.05）。W e s t e r n b l o t 结果：与对照组比较,Bax、Ca s p a s e-3在丙泊酚组显著表达，差异具有统计学意义（P0.05），而Bcl-2、CREB表达明显减少，差异具有统计学意义（P0.05）；与丙泊酚组比较,Bax、C a s p a s e-3在电针组表达明显减少,差异具有统计学意义（P0.05）,相反,Bcl-2、CREB在电针组显著表达，差异具有统计学意义（P0.05）。结论：“百会”“大椎”穴电针预处理可以有效减轻反复丙泊酚麻醉诱发的大鼠学习记忆障碍,其机制可能与上调pCREB、CREB蛋白以及调控凋亡相关因子的表达，进而抑制神经元凋亡，改善学习记忆，发挥神经保护作用。关键词电针预处理；丙泊酚；凋亡；学习记忆障碍中图分类号：R246.6D0I:10.19917/ki.1005-0779.023161Electro-Acupuncture Preconditioning at DU20 and DU14 Attenuating Learningand Memory Impairment Induced by Repeated Propofol Anesthesia in RatsFAN Shunqin*2,WANG Xijun,WEI Songl,LU Sishil,CUO Yongrong(1.Guangxi International Zhuang Medical Hospital Afiliated to Guangxi University of Chinese Medicine,Nanning 530201,China;2.Guangxi University of Chinese Medicine,Nanning 530200,China)Abstract Objective:To observe the effect of electro-acupuncture(EA)preconditioning at Baihui(DU20)and Dazhui(DU14)on reducing learning memory impairment induced by repeated propofol anesthesiain rats.Methods:The model of learning and memory impairment was induced by 5 consecutive days of propofolanesthesia,and EA pretreatment was given for 5 consecutive days before propofol administration.The learningand memory ability was evaluated by Morris water maze.The amount and morphological change of hippocampalneurons were observed by HE staining.The protein expression of pCREB in the Dentate Gyrus region of基金项目：广西中医药管理局项目，编号：ZGJ202173；广西国际壮医医院高层次人才引进基金，编号：GZ2021RC009；广西中医药大学研究生教育创新计划，编号：YCSY2023059。作者简介：范舜钦（1997），男，2 0 2 1级麻醉学专业硕士研究生，研究方向：围术期器官功能保护。通讯作者：王喜军（197 3-），男，教授，硕士研究生导师，研究方向：围术期重要器官功能保护。文献标识码：A针灸临床杂志2 0 2 3年第39卷第8 期hippocampus was measured by immunofluorescence staining.The protein expressions of Bcl-2,Caspase-3,Bax and CREB were determined by Western Blotting.Results:In terms of Morris water maze results,comparedwith those in the control group,the number of platform crossing,the swimming time and the percentage of pathsin the target quadrant were significantly reduced in the propofol group(P0.05);compared with those in thepropofol group,the number of platform crossing,the swimming time and the percentage of paths in the targetquadrant were significantly increased in the EA group(P0.05).In terms of the HE results,compared withthose in the control group,the number of cell layers in the hippocampal DG area decreased and the cell densitydecreased in the propofol group;compared with those in the propofol group,the number of neuronal cell layersin the hippocampal DG area significantly recovered and the cell density significantly increased in the EA group.In terms of the immunofluorescence results,compared with that in the control group,the intergratedfluorescence intensity of pCREB protein in the hippocampal region was significantly reduced in the propofolgroup(P0.05);compared with that in the propofol group,the intergrated fluorescence intensity of pCREBprotein in the hippocampal region was significantly increased in the EA group(P0.05).In terms of theWestern Blotting results,compared with those in the control group,the expressions of Bax and Caspase-3 weresignificantly increased(P0.05),whereas the expressions of Bcl-2 and CREB were significantly decreasedin the propofol group(P 0.05);compared with those in the propofol group,the expressions of Bax andCaspase-3 were significantly decreased(P 0.05),and the expressions of Bcl-2 and CREB weresignificantly increased in the EA group(P0.05).Conclusion:Electro-needling DU20 and DU14 caneffectively reduce learning and memory impairment induced by repeated propofol anesthesia in rats,and themechanism may be related to up-regulating the proteins of pCREB and CREB and the expression ofapoptosis-related factors,which in turn inhibits neuronal apoptosis,improves learning and memory ability,and exerts neuroprotective effects.Key words Electro-acupuncture preconditioning;Propofol;Apoptosis;Learning and memory;impairment79.丙泊酚作为临床使用面广、安全系数较高的静脉麻醉药,因其诱导快、半衰期短和苏醒快等特点而广泛应用于婴幼儿麻醉诱导及维持。研究发现,反复丙泊酚麻醉会产生发育神经毒性，可引起发育期大鼠大脑神经元调亡，激活大脑炎症反应、氧化应激反应等，最终导致大鼠学习记忆能力的损害。因此,麻醉前采取必要的干预措施对预防或减轻学习记忆损伤具有积极意义。电针是传统针灸与现代电刺激的完美结合产物,具有简便、安全与稳定等优点，研究表明2 ，电针“百会”“大椎”穴可减轻神经元调亡，发挥脑保护效应。基于此，本研究观察“百会”“大椎”穴电针预处理对反复丙泊酚麻醉诱导的大鼠学习记忆能力损害的影响，为围术期麻醉安全提供参考。1材料与方法1.1实验动物广西医科大学实验动物中心（实验动物许可证号：SCXK桂 2 0 140 0 0 2）提供3周龄雄性健康SD大鼠6 0 只，体质量55 7 0 g。SD 大鼠分组完成后饲养于温度2 3 2 5，湿度52%56%，可自由进食水的清洁动物房。广西中医药大