目的 研究针刺对脑缺血再灌注损伤(cerebral ischemia reperfusion injury,CIRI)大鼠海马区环状RNA_009775 表达的影响,探索针刺对CIRI可能的作用机制.方法 将 39只SD大鼠随机分为假手术组、模型组、针刺组,每组 13只.除假手术组外,其余两组均采用线栓法制备CIRI模型.造模后,针刺组大鼠捆绑固定行针刺干预,间隔 12h进行 1次,共 7次;假手术组与模型组大鼠仅捆绑不针刺.Garcia评分法、TTC染色法评估针刺疗效;基因芯片技术筛选针刺组/模型组、模型组/假手术组核心共同差异表达circRNA;qRT-PCR法对芯片筛选结果进行验证;TargetScan软件预测核心circRNA的下游miRNA及mRNA,构建核心circRNA的调控网络,并对mRNA进行基因本体功能富集分析;Western blot法验证神经元核抗原抗体(neuron specific nuclear protein,NEUN)表达量.结果 干预后,与假手术组相比,模型组大鼠Garcia评分、NEUN和circRNA_009775 表达量显著降低(P＜0.01),脑梗死面积比显著升高(P＜0.01);与模型组相比,针刺组脑梗死面积比显著降低(P＜0.01),Garcia评分、NEUN和circRNA_009775 表达量显著升高(P＜0.05 或P＜0.01).芯片筛选核心共同差异表达circRNA,经qRT-PCR验证circRNA_009775 表达趋势与芯片结果一致,结合靶关系预测,构建了含有 1个circRNA、12个miRNA、31个mRNA的调控网络.基因本体功能富集分析显示mRNA主要涉及神经系统发育、神经元生成等生物过程.结论 针刺具有改善CIRI大鼠神经功能缺损、降低脑梗死面积、部分减轻海马区神经元损伤的作用,其机制可能与调控circRNA_009775 介导的潜在调控网络、促进神经系统发育、神经元生成等生物过程有关.

目的:探讨针刺对脑缺血再灌注损伤(CIRI)大鼠海马区细胞凋亡的影响及机制.方法:SD大鼠共75只,随机分为假手术组(sham)、脑缺血再灌注损伤模型组(CIRI)和针刺治疗组(CIRI+AC),利用大脑中动脉栓塞法制备CIRI大鼠模型,CIRI+AC组大鼠接受针刺"大椎"、"水沟"、"百会"穴治疗.Garcia评分法检测大鼠神经功能,TTC染色法检测大鼠脑梗死面积,免疫组织化学染色检测海马区caspase-3阳性细胞数量,Western Blot检测海马区caspase-9表达,TUNEL检测细胞凋亡.结果:针刺前,CIRI组和CIRI+AC组神经功能评分较sham组显著降低(P＜0.01);针刺24、72 h后,CIRI+AC组神经功能评分较CIRI组显著升高(P＜0.01),脑梗死面积比缩小(P＜0.05),患侧脑组织细胞凋亡率降低(P＜0.01);与针刺24 h后比较,72 h后CIRI组患侧脑组织细胞凋亡率及海马区caspase-3阳性细胞数量增加(P＜0.05),CIRI+AC组患侧脑组织细胞凋亡率及海马区caspase-3阳性细胞数量明显降低(P＜0.05).与CIRI组比较,针刺24 h后CIRI+AC组患侧海马区caspase-9表达显著升高(P＜0.01),针刺72 h后caspase-9表达显著降低(P＜0.01).结论:持续针刺可改善大鼠神经功能和细胞凋亡情况,且针刺对CIRI大鼠海马区细胞凋亡的抑制具有时效性.

ObjectiveTo explore the mechanisms of acupuncture against cerebral ischemia/reperfusion injury （CIRI） through observing the expression of circular RNA HDAC2 （circHDAC2） in the hippocampus of rats.MethodsSD rats were randomly divided into sham-operation， model and acupuncture groups， with 13 rats in each group. The rat model of CIRI was established by middle cerebral artery occlusion. In the acupuncture group， acupuncture was delivvered at “Dazhui” （GV14）， “Shuigou” （GV26） and “Baihui” （GV20）， and the needles were retained for 30 min each time and acupuncture was conducted once every 12 h for a total of 7 sessions. Before and after intervention， using modified Garcia scale， the neurological function of the rats were evaluated， and TTC staining was employed to determine the cerebral infarct area. Gene chip technology was used to screen the circRNAs with differential expressions in the ischemic hippocampus， and the circRNAs with co-differential expression （co-DE circRNAs） in the model group/sham-operation group， and the acupuncture group/model group separately. Among those circRNAs， the core circRNAs were screened according to P value， fold change （FC） and gene ontology （GO） analysis； and their expressions in the ischemic hippocampus were determined using quantitative real-time PCR （qPCR）. Based on the verification results， a competing endogenous RNA （ceRNA） prediction network was constructed. The expression levels of microRNA （miRNA） and mRNA with high node centrality in the prediction network were validated by qPCR.ResultsBefore intervention， compared with the sham-operation group， the modified Garcia score of each modeling group decreased （P<0.01）. After intervention， the modified Garcia score was reduced and the cerebral infarct area ratio increased （P<0.01） in the model group when compared with the sham-operation group. In the acupuncture group， the modified Garcia score was higher and the cerebral infarct area ratio lower （P<0.01） than those of the model group. The microarray results of gene chip showed that 16 co-DE circRNAs were down-regulated in the model group and up-regulated in the acupuncture group， and 7 co-DE circRNAs up-regulated in the model group and down-regulated in the acupuncture group. The core circHDAC2 and circNTRK2 were screened according to P value， FC and the enrichment number of GO entries. QPCR results showed that， compared with the sham-operation group， the expression of circHDAC2 and circNTRK2 of the ischemic hippocampal tissue was down-regulated in the model group （P<0.01）； and that of circHDAC2 and circNTRK2 up-regulated in the acupuncture group when compared with the model group （P<0.01）. The relevant ceRNA regulatory network was constructed for circHDAC2 and the prediction results showed that the regulatory networks contained 12 miRNAs and 31 mRNAs. Results of verifying miRNA with high network node centrality and mRNA relevant with nerve regulation showed that， when compared with the sham-operation group， the expression levels of miR-29a， miR-29b and the solute carrier family 30 member 3 （SLC30A3） mRNA in the ischemic hippocampus were down-regulated （P<0.01）； and those of miR-3065 and mercaptopyruvate sulfurtransferase （MPST） up-regulated （P<0.01） in the model group. Compared with the model group， the expressions of miR-29a， miR-29b and SLC30A3 mRNA in the ischemic hippocampus were up-regulated （P<0.01，P<0.05）， while that of miR-3065 down-regulated （P<0.05） in the acupuncture group.ConclusionAcupuncture significantly improves the neurological function and reduces the cerebral infarct area in CIRI rats， which may be related to the regulation of hippocampal circHDAC2/miR-3065/SLC30A3 axis.

探讨"心胀咽痛,刺太冲必除之"理论依据,扩展太冲穴临床应用.从心与肝生理病理、气血调节联系、经络循行理论以及标本根结理论四方面分析,得出肝疏泄失常致心主血脉、心主神志功能异常,故治心脏疾患需先调肝清其源;肝与心,气与血的调节相互影响,调气为主,调血为辅,不可忽视肝调节气机对维持其他脏腑生理功能的重要性;太冲穴为肝经的原穴,针刺其可治疗胸腹部疾病;针刺太冲可调理气机不畅,血行不利所致心神疾患,故"心胀咽痛,刺太冲必除之"理论可指导临床应用太冲穴治疗胸闷心烦、短气、咽痛、卧不安等.

目的 探讨针刺对脑缺血再灌注损伤(CIRI)大鼠脑保护作用与海马组织差异表达核心基因、转录网中信使RNAs(mRNAs)功能之间的联系,为针刺调控环状RNAs(circRNAs)抗CIRI后神经损伤研究提供参考.方法 将54只SD大鼠随机分为假手术组、模型组和针刺组,每组18只,后两组采用改良Longa线栓法制备大脑中动脉阻塞再灌注模型,假手术组仅剥离血管,不插线栓.干预期内针刺组捆绑于鼠板并针刺"大椎""水沟""百会"穴,均匀捻转刺激1 min,频率为90次/min,15 min后行针1次,共留针30 min,每12 h干预1次,共7次.假手术组、模型组捆绑于鼠板不针刺.改良加西亚(Garcia)评分法检测大鼠神经功能状况,2,3,5-氯化三苯基四氮唑(TTC)染色法观察脑梗死面积;通过基因芯片技术筛选缺血侧海马组织差异表达基因,整理出模型组/假手术组、针刺组/模型组共同差异表达的环状RNAs(co-DE circRNAs),根据P值、差异倍数(FC)、基因本体(GO)数据库条目富集数量筛选出高表达的核心co-DE circRNAs,分析核心co-DE circRNAs来源基因的GO功能;统计缺血侧海马CA1区尼氏染色阳性细胞数;通过实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法验证核心co-DE circRNAs表达量,选取表达趋势与芯片结果一致的circRNAs进行竞争性内源RNA(ceRNA)网络预测,对预测网络中的mRNAs进行GO功能富集分析.结果 干预前,与假手术组比较,各造模组改良Garcia 神经功能评分显著降低(P<0.05);干预后,与假手术组比较,模型组脑梗死面积比显著升高,改良 Garcia 神经功能评分和CA1区尼氏染色阳性细胞数显著降低(P<0.05);与模型组比较,针刺组脑梗死面积比显著降低,改良Garcia 神经功能评分和CA1区尼氏染色阳性细胞数显著升高(P<0.05);组内干预前后比较,针刺组改良Garcia 神经功能评分显著升高(P<0.05).芯片结果显示,模型组下调/针刺组上调、模型组上调/针刺组下调的co-DE circRNAs为23个,根据FC>1.25和GO数据库条目富集数量筛选出核心基因为环状RNA组蛋白去乙酰化酶2基因(circHDAC2)、环状RNA神经营养受体酪氨酸激酶2基因(circNTRK2);GO数据库分析显示,二者富集于神经元的产生、神经系统发育、神经元分化等条目.验证其表达量发现,与假手术组比较,模型组大鼠缺血侧海马组织circHDAC2、circNTRK2 表达量均显著下调(P<0.05);与模型组比较,针刺组大鼠缺血侧海马组织circHDAC2、circNTRK2表达量均显著上调(P<0.05).circHDAC2表达趋势与芯片结果一致,由此对circHDAC2 构建相关 ceRNA调控网络,调控网络中mRNAs的GO分析主要富集在神经元的产生、神经元分化的调控、轴突引导、神经系统发育、神经元死亡的调节等条目.结论 针刺可改善CIRI大鼠改良 Garcia神经功能评分与脑梗死面积比,降低缺血侧海马组织的神经元损伤,其作用可能与调控缺血侧海马组织circHDAC2表达以及激发调控网络中mRNA的神经元产生、神经元分化、神经元细胞体功能等有关.

Objective To explore the regulation of acupuncture on the expression of circRNAs in the ischemic hippocampus of rats with cerebral ischemia-reperfusion injury, so as to provide experimental theoretical basis for the treatment of cerebral ischemia with acupuncture. Methods Twenty four healthy adult male SD rats were randomized into acupuncture group, model group and sham-operated group, with 8 rats in each group. The middle cerebral artery occlusion/reperfusion（MCAO/R） model of rats was established by thread embolism. After the rats in the acupuncture group were bound, the points of Dazhui（GV14）, Baihui（GV20）and Renzhong（GV26） were selected for acupuncture. The needles were manipulated every 15 min and were retained for 30 min.The treatment was performed every 12 h, 7 times in a row. The model group and the sham-operated group were only bound without any treatment. The neurological function was assessed by Garcia score before and 72 h after intervention. At 72 h after intervention,the area ratio of cerebral infarction was observed by TTC staining, and the expression profile of circRNAs in the ischemic hippocampus was detected by circRNAs gene chip. Results（1） Before intervention: Compared with the sham-operated group, the score of neurological function impairment in the acupuncture group and the model group dropped significantly（P ＜0.01）. After intervention: Compared with sham-operated group, the score of neurological function impairment in the acupuncture group and the model group decreased significantly, whereas the area ratio of cerebral infarction increased（P ＜0.01）. Compared with the model group, the score of neurological function impairment in the acupuncture group went up while the area ratio of cerebral infarction went down（P＜0.05）.（2） CircRNA chip analysis: Compared with the sham-operated group, there were 603 differential genes in the model group, including 315 down-regulated circRNAs and 288 up-regulated circRNAs. Compared with the model group, there were51 differential genes in the acupuncture group, including 18 down-regulated circRNAs and 33 up-regulated circRNAs. There were23 circRNAs under co-regulation. Compared with the sham-operated group, the function of the differentially expressed circRNAs showed changes in synapses between neurons and cell morphology, which were mainly attributed to ErbB signaling pathway, MAPK signaling pathway, cAMP signaling pathway, etc. Compared with the model group, the function of the differentially expressed circRNAs in the acupuncture group showed changes in neurotrophin binding, H3 histone acetyltransferase complex, and retrograde axonal transport, which were mainly involved in MAPK signaling pathway, neurotrophin signaling pathway, Hippo signaling pathway,etc. The functions of co-regulation include neuron projection, protein binding, nervous system development, etc. The signaling pathway of co-regulation includes MAPK signaling pathway. Conclusion Acupuncture at Dazhui（GV14）, Baihui（GV20） and Renzhong（GV26） can improve the score of neurological function impairment and reduce the area ratio of cerebral infarction in rats with cerebral ischemia-reperfusion injury. The mechanism may be related to regulating the expression of multiple circRNAs, activating multiple functions and multiple signaling pathways.

目的 探讨霉帕霉素(RAPA)作用下针刺激活Ⅲ型PI3K-Beclin1通路抗脑缺血再灌注后细胞自噬的研究,为临床针刺治疗脑血管疾病提供一定的实验依据.方法 SD健康55只大鼠,随机分为假手术组11只、造模组44只.造模组参考线栓法制备大鼠CIRI模型.造模成功后再随机分为模型组、针刺组、模型+RAPA组和针刺+RAPA组.大鼠生命体征稳定后,假手术组、模型组、模型+RAPA组只捆绑固定不针刺;针刺组和针刺+RAPA组捆绑固定并针刺"大椎"、"人中"、"百会"穴,针后行捻转手法1 min(频率90次/min),每15 min行捻转手法1 min,留针30 min,每12h针刺1次,共7次.干预前及72h后进行改良Garcia评分观察大鼠神经功能损伤程度,TTC染色法观察大鼠脑梗死面积比,Western Blot法检测Ⅲ型PI3K、Beclin-1、LC3Ⅱ/Ⅰ、Lamp2、P62蛋白的表达水平,透射电镜观察自噬小体.结果 ①改良Garcia神经功能评分:干预前,与假手术组比较,各造模组改良Garcia评分显著降低(P＜0.01).干预后,与假手术组比较,各造模组改良Garcia评分明显降低(P＜0.01);与模型组比较,针刺组、模型+RAPA组和针刺+RAPA组改良Garcia评分明显升高(P＜0.01).组内干预前后比较,针刺组、模型+RAPA组和针刺+RAPA组较干预前改良Garcia评分上升(P＜0.01).②脑梗死面积比:与假手术组比较,各造模组脑梗死面积比明显升高(P＜0.01);与模型组比较,针刺组、模型+RAPA组和针刺+RAPA组大鼠脑梗死面积比明显降低(P＜0.01);与针刺组、模型+RAPA组比较,针刺+RAPA组大鼠脑梗死面积比降低(P＜0.01).③Ⅲ型PI3K、Beclin-1、LC3Ⅱ/Ⅰ、Lamp2、P62表达水平:与假手术组比较,模型组Ⅲ型PI3K、Bec-lin-1、LC3Ⅱ/Ⅰ、Lamp2表达水平显著降低,P62表达水平显著增高(P＜0.01);与模型组比较,针刺组、模型+RAPA组、针刺+RAPA组Ⅲ型PI3K、Beclin-1、LC3Ⅱ/Ⅰ、Lamp2表达显著升高,P62表达水平降低(P＜0.01);与模型+RAPA组比较,针刺+RAPA组LC3Ⅱ/Ⅰ表达水平升高,P62表达水平降低(P＜0.01).④大鼠海马神经元自噬超微结构:假手术组神经元细胞呈轻微水肿,胞膜、核膜清晰、完整,Ly、SL、ASS、AP个别存在.模型组神经元细胞水肿较严重,大多数线粒体局部基质溶解,嵴减少;SL少量存在,未见典型自噬结构.模型+RAPA组神经元细胞呈中度水肿,细胞膜完整,观察到Ly、ASS数量较多.针刺组、针刺+RAPA组神经元细胞整体结构尚可,未见明显损伤,胞膜、核膜清晰、完整,Ly、SL存在,可见ASS、AP.结论 针刺激活Ⅲ型PI3K-Beclin1通路,增加自噬标志物LC3Ⅱ/Ⅰ、Lamp2的表达,降低溶酶体标志物P62的表达,以激活适度自噬减轻CIRI.

目的:探讨针刺对脑缺血再灌注损伤(CIRI)大鼠脑组织的保护作用,观察针刺对CIRI大鼠缺血侧海马组织环状RNAs(circRNAs)差异表达的影响,并对其进行基因本体(GO)分析.方法:6～8周龄SD大鼠54只,运用随机数字法随机分为造模组和假手术组(sham组),造模成功后再随机分为模型组(model组)和针刺组(AC组),每组18只.采用改良Longa线栓法制备大脑中动脉闭塞再灌注(MCAO/R)模型,激光散斑成像仪监测造模前、MCAO手术后及再灌注后脑血流量,假手术组只剥离血管,不插入线栓;干预期间模型组和假手术组只捆绑不针刺,针刺组捆绑+针刺.采用改良加西亚(Garcia)评分法对神经功能进行评定,TTC染色法检测脑梗死面积,Western blot法检测神经元核抗原(NeuN)的表达,尼氏染色法观察缺血侧海马组织神经元损伤程度,基因芯片微阵列分析筛选出缺血侧海马组织差异表达的circRNAs,并对模型组/假手术组、针刺组/模型组共同差异表达circRNAs的来源基因进行GO分析.结果:与假手术组比较,模型组大鼠脑梗死面积比显著升高(P<0.01),Garcia神经功能评分、NeuN表达量和海马CA1区尼氏染色阳性细胞数显著降低(P<0.05或P<0.01);与模型组比较,针刺组脑梗死面积比显著降低(P<0.01),神经功能评分、NeuN表达量和尼氏染色阳性细胞数显著升高(P<0.05或P<0.01).芯片筛选结果显示,与假手术组比较,模型组上调的circRNAs个数为288,下调个数为315;与模型组比较,针刺组上调的差异表达circRNAs个数为33,下调个数为18(FC>1.25,P<0.05);其中模型组/假手术组、针刺组/模型组共同差异表达的circRNAs个数为23个;GO分析显示共同差异表达circRNAs的来源基因功能涉及神经系统发育,神经元的产生、发育、分化及投射,头部、大脑及海马的发育,突触的形成、发育、延伸及运输等.结论:CIRI大鼠缺血侧海马组织circRNAs在造模后及针刺干预后均存在差异表达.针刺能显著改善CIRI大鼠的神经功能和脑梗死面积,减轻海马组织神经元损伤,其机制可能与针刺调控缺血侧海马组织多种circRNAs的差异表达及激发其促进神经元发育分化、抗神经损伤等功能有关.

探讨歌赋"胸结身黄,取涌泉而即可"的作用机制及选穴依据,以拓展涌泉穴在治疗"胸结身黄"方面的临床应用.从"经脉所过,主治所及"的经络理论、"病在脏者,取之井"的脏腑理论、"病在上者,取之下"的标本根结理论、"治病求于本"等4个方面进行探析.总结出位于四肢末端根部的井穴涌泉穴具有较强的治疗胸腹部脏腑疾病和清热、开郁退黄、导热下行的功用,体现了疏通经络、调和阴阳、扶正祛邪等具有中医特色的针灸治病原理,以及循经选穴、上下配穴的针灸选穴规律.通过深入挖掘其理论内涵以及对"胸结身黄"病因病机的认识,为其临床治疗拓宽了思路,提高了临床疗效.

目的:利用大脑中动脉闭塞/再灌注(MCAO/R)法制备大鼠脑缺血再灌注损伤(CIRI)模型,研究针刺对CIRI模型大鼠脑组织中Ⅲ型磷脂酰肌醇-3-羟激酶(PI3K-Ⅲ)/Beclin-1通路的影响.方法:利用MCAO/R方法制备SD大鼠CIRI模型,分别给予针刺和(或)自噬抑制剂3-甲基腺嘌呤(3-MA)进行治疗.利用采用Gar-cia评分法检测大鼠神经功能,TTC染色法检测大鼠脑梗死面积,Western Blot法检测海马组织PI3K-Ⅲ、Beclin-1和B淋巴细胞瘤-2基因(Bcl-2)蛋白的表达水平,TUNEL法检测细胞凋亡.结果:MCAO/R手术后大鼠神经功能评分降低(P＜0.01),TTC染色观察到脑梗死,同时海马区PI3K-Ⅲ、Beclin-1和Bcl-2表达显著下调(P＜0.01),脑皮质细胞凋亡增多(P＜0.01).针刺治疗不但提高了大鼠神经功能评分(P＜0.01),并且减少了脑梗死面积(P＜0.01);同时显著上调了海马组织中PI3K-Ⅲ、Beclin-1和Bcl-2表达(P＜0.01),并减少了脑细胞凋亡(P＜0.01).3-MA阻断了针刺的治疗作用,而且使脑损伤更加严重.结论:针刺可改善CIRI大鼠神经功能并减轻神经损伤,其机制可能与激活PI3K-Ⅲ/Beclin-1通路增加细胞自噬并减少细胞凋亡有关.

目的 从海马体基因表达方面探讨针刺大椎、百会、水沟穴治疗脑缺血再灌注损伤的可能作用机制.方法 39只SD大鼠随机分为假手术组、模型组、针刺组各13只.模型组和针刺组大鼠使用线栓法制备大鼠脑缺血再灌注损伤模型.造模后,针刺组大鼠捆绑后针刺"百会""水沟""大椎"穴,留针30min,每15 min捻转1次,平补平泻,共针刺7次,每次间隔12 h;假手术组、模型组只捆绑不针刺.于针刺前、针刺后进行神经功能缺损评分;针刺后TTC染色观察脑梗死情况,采用circRNAs微阵列芯片分析大鼠缺血侧海马区circRNAs差异表达,并用qRT-PCR法验证差异基因的表达.结果 针刺前,与假手术比较,针刺组、模型组神经功能评分显著降低(P＜0.01);针刺后,针刺组神经功能评分较本组针刺前提高,亦高于模型组(P＜0.05).与模型组比较,针刺组脑梗死面积比下降(P＜0.05).CircRNAs微阵列芯片检测结果显示,模型组/假手术组差异表达的circRNAs共215个,其中上调59个,下调156个;针刺组/假手术组差异表达的circRNAs共5个,其中上调1个,下调4个;针刺组/模型组差异表达circRNAs 2个,均上调.模型组/假手术组与针刺组/模型组共表达的差异基因有2个,分别为circ-011989和circ-009775.选取共表达差异基因Ilog2 FCI≥1.5最高的差异基因circ-011989进行验证,结果显示,与假手术组比较,模型组大鼠海马区circ-011989表达量减少(P＜0.05);与模型组比较,针刺组大鼠海马区circ-011989表达量增加(P＜0.05).结论 针刺"大椎""百会""水沟"穴可改善脑缺血再灌注损伤模型大鼠神经功能,减小脑梗死面积,其机制可能与调控缺血侧海马区circ-011989的表达有关.

OBJECTIVE To observe the effects of acupuncture on the expression of type Ⅲ phosphatidylinositol 3-hydroxykinase (PI3K) and Beclin-1 in hippocampus of rats with cerebral ischemia/reperfusion injury (CI/RI), so as to explore the mechanism of acupuncture in regulating type Ⅲ PI3K pathway to activate autophagy in the hippocampal neurons of CI/RI rats. METHODS SD rats were randomly divided into sham operation group (n=11) and operation group. Then after successful modeling, rats in the operation group were randomly divided into model, acupuncture, model+3-MA and acupuncture+3-MA groups, with 11 rats in each group. The model of CI/RI was established by occlusion of the middle cerebral artery. Rats in the model+3-MA and acupuncture+3-MA groups were injected with 3-MA (400 nmol/ 5 μL) 5 μL into the lateral ventricle 30 min before reperfusion. Rats in the acupuncture and acupuncture+3-MA groups were punctured with filiform needles at "Dazhui" (GV14), "Shuigou" (GV26) and "Baihui" (GV20) and stimulated manually once every 15 min. The acupuncture intervention was conducted for 30 min each time, once every 12 h for a total of 7 times. The degree of neurological impairment was evaluated 2 h after reperfusion and after intervention by Garcia score. After intervention, the percentage of cerebral ischemic area was observed by TTC staining, the protein expression levels of type Ⅲ PI3K, Beclin-1, microtubule-associated protein 1 light chain 3B (LC3B), lysosome associated membrane protein 2 (Lamp2) and P62 in ischemic hippocampal tissue were detected by Western blot, the ultrastructure of neurons in ischemic hippocampus was observed by transmission electron microscopy (TEM). RESULTS Compared with the sham operation group, the Garcia score was decreased (P<0.01), the percentage of cerebral ischemic area was increased (P<0.01), the expression levels of type Ⅲ PI3K, Beclin-1, LC3B-Ⅱ/Ⅰ, Lamp2 proteins were decreased (P<0.01), and the expression level of P62 protein was increased (P<0.01) in ischemic hippocampal tissue in the model group. Compared with the model group, the Garcia score was increased (P<0.01), the percentage of cerebral ischemic area was decreased (P<0.01), the expression levels of type Ⅲ PI3K, Beclin-1, LC3B-Ⅱ/Ⅰ, Lamp2 proteins were increased (P<0.01, P<0.05) and the expression level of P62 was decreased (P<0.01) in ischemic hippocampal tissue in the acupuncture group； the percentage of cerebral ischemic area was increased (P<0.05), the expressions of type Ⅲ PI3K and Beclin-1 were decreased (P<0.01) and the expression level of P62 protein was increased (P<0.05) in ischemic hippocampal tissue in the mo-del+3-MA group. Compared with the model +3-MA group, the Garcia score was increased (P<0.05), the percentage of cerebral ischemic area was decreased (P<0.01), the expression levels of type Ⅲ PI3K, Beclin-1, LC3B-Ⅱ/Ⅰ in ischemic hippo-campal tissue were increased (P<0.01, P<0.05) in the acupuncture+3-MA group. Compared with the acupuncture group, the Garcia score was decreased, the percentage of cerebral ischemic area was increased (P<0.05, P<0.01), the expression levels of type Ⅲ PI3K, Beclin-1, Lamp2 proteins were decreased (P<0.01, P<0.05) and P62 protein was increased (P<0.05) in ischemic hippocampal tissue in the acupuncture+3-MA group. The results of TEM showed that the edema of neurons was heavier, and few hypolysosomes existed in the model group; there was no obvious damage to neuronal structure, intracellular matrix was abundant, and a few lysosomes existed in the acupuncture group; the neuronal cells had mild edema and primary lysosomes were present in the acupuncture +3-MA group. CONCLUSION Acupuncture can improve the symptoms of neurological impairment and reduce the percentage of cerebral ischemic area in rats with CI/RI. The mechanism may be related to regulating type Ⅲ PI3K/Beclin-1 pathway, up-regulating the expressions of autophagy related factors LC3B-Ⅱ and Lamp2, and down-regulating the expression of P62.

目的 探讨针刺对脑缺血再灌注损伤(cerebral ischemia reperfusion injury,CIRI)大鼠的脑保护作用及缺血侧海马组织铁死亡的影响.方法 65只健康SD大鼠随机分成假手术组、模型组、针刺组、药物组,药物组5只,其他每组20只.线栓法制备大鼠脑缺血模型,2 h后拔出线栓约10 mm,建立CIRI模型.大鼠生命体征稳定后,药物组采用依达拉奉水溶液(5 mg/kg)腹腔注射后捆绑30 min;假手术组、模型组注射同等药物剂量生理盐水,并捆绑30 min;针刺组注射同等药物剂量生理盐水并选取"人中""大椎""百会"穴进行捆绑针刺,留针30 min.均1次/12 h,共7次.采用改良Garcia评分观察大鼠神经功能损伤程度;TTC染色检测脑梗死面积;铁离子、活性氧(reactive oxygen species,ROS)及丙二醛(malondialdehyde,MDA)试剂盒检测海马组织铁离子、ROS及MDA含量.结果(1)干预前,与假手术组相比,其余各组改良Garcia评分显著降低(P<0.01).干预后,与假手术组相比,模型组改良Garcia神经功能评分明显降低,脑梗死面积显著扩大(P<0.01);与模型组相比,针刺组、药物组改良Garcia神经功能评分升高,脑梗死面积缩小(P<0.05,P<0.01).与干预前相比,针刺组、药物组改良Garcia神经功能评分升高(P<0.05).(2)与假手术组相比,模型组海马组织铁离子、ROS及MDA含量均明显增加(P<0.01);与模型组相比,针刺组海马组织铁离子、ROS及MDA含量均明显降低(P<0.01).结论 针刺可以抑制CIRI后大鼠神经细胞铁死亡,发挥脑保护作用,可能与调节神经细胞铁离子代谢、减轻海马组织铁离子集聚、降低脂质过氧化水平有关.

明确《行针指要歌》中"或针吐,中脘气海膻中补"治疗反胃吐食,噎膈呃逆等脾胃疾病的作用机制及选穴依据,为临床治疗脾胃疾患提供参考;从三焦气机理论、经络腧穴理论及治未病理论探析其理论依据.膻中穴为八会穴之气会,与全身气机紧密相关;中脘穴为八会穴之腑会、胃之募穴,可调脾胃气机;气海穴为元气生发汇聚之所,可治一身气疾;三穴同用可顾护胃气,使脾升胃降,气机通畅,针对反胃吐食、噎膈呃逆、慢性胃炎等脾胃疾病有较好的疗效.并引用杨继洲和石学敏的古今医案,论证可通过调理气机来治疗临床脾胃病症;根据以上理论探讨,"或针吐,中脘气海膻中补"中选穴及配穴方法切实可行,可灵活运用于临床上的脾胃疾患等.