眼针对脑缺血再灌注损伤大鼠神经血管单元细胞凋亡的影响  

Eye-acupuncture intervention reduces cerebro-cortical apoptosis of neurovascular unit in cerebral ischemia-reperfusion injury rats

作　　者：宓丹[1] 王德成 潘冲 MI Dan;WANG De-cheng;PAN Chong(Department of Emergency,the Second Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110034,China;Department of Laboratory,the Second Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110034,China)

机构地区：[1]辽宁中医药大学附属第二医院急诊科,沈阳110034 [2]辽宁中医药大学附属第二医院检验科,沈阳110034

出　　处：《针刺研究》2019年第1期13-18,共6页Acupuncture Research

基　　金：辽宁省科技厅研究项目(No.2014020162)

摘　　要：目的:观察眼针对脑缺血再灌注损伤大鼠脑组织微血管内皮细胞、星形胶质细胞、神经元凋亡及大脑皮质Bad、Bcl-xL表达的影响,探讨眼针对脑缺血再灌注损伤大鼠神经血管单元保护作用的机制。方法:SD大鼠随机分为空白组、假手术组、模型组(3、24、72h组),眼针组(3、24、72h组),每组12只。采用改良线栓法制备脑缺血再灌注损伤大鼠模型。眼针组针刺"肝区""肾区""上焦""下焦",每次20min,每12h治疗1次。造模后3、24、72h采用Western blot法检测缺血侧大脑皮质Bad、Bcl-xL蛋白的表达;造模后72h采用凋亡免疫荧光双标记法检测神经元、微血管内皮细胞、星形胶质细胞凋亡情况。结果:与空白组比较,脑缺血再灌注72h后模型大鼠缺血侧大脑皮质的神经元、微血管内皮细胞、星形胶质细胞均可见不同程度的凋亡(P<0.01)。眼针72h组与模型72h组比较,神经元、微血管内皮细胞、星形胶质细胞凋亡数减少(P<0.01)。与空白组比较,模型组3、24、72h大脑皮质Bad蛋白表达上调(P<0.01);与同时间点模型组比较,眼针24h组、眼针72h组Bad蛋白表达减少(P<0.05)。与空白组比较,模型3h组、24h组、72h组大脑皮质Bcl-xL蛋白表达上调(P<0.01);与同时间点模型组比较,眼针3h组、24h组、72h组Bcl-xL蛋白表达上调(P<0.01)。结论:眼针治疗可以通过减少Bad蛋白的表达,促进Bcl-xL的表达,减少脑缺血后神经血管单元的主要成分神经元、微血管内皮细胞、星形胶质细胞的凋亡,从而对脑缺血后的神经血管单元起到保护作用。Objective To observe the effect of eye-acupuncture intervention on cerebro-cortical apoptosis of microvascular endothelial cells,neurons and astrocytes(main components of neurovascular unit)and the expression of Bad(an apoptosis promoter)and B-cell lymphoma-extra large(Bcl-xL)proteins in focal cerebral ischemia-reperfusion injury(CIRI)rats,so as to explore its mechanisms underlying improvement of CIRI.Methods SD rats were randomly divided into control,sham-operation,model 3 h,model 24 h,model 72 h,eye-acupuncture 3 h,eye-acupuncture 24 hand eye-acupuncture 72 hgroups(n=12 in each group).The CIRI model was established by middle cerebral artery occlusion/reperfusion(MCAO/R).Eye-acupuncture was applied to bilateral"Gan"(Liver)regions,"Shen"(Kidney)regions,"Shangjiao"(Upper-energizer)and"Xiajiao"(Lower-energizer)for 20 min,once 3 hand every 12 hafter modeling.The expression levels of Bad and Bcl-xL in the ischemic cerebral cortex tissue were detected by Western blot.The apoptotic neurons,microvascular endothelial cells and astrocytes were assayed by immunofluorescence double labeling(Nestin/TUNEL,CD34/TUNEL and glial fibrillary acidic protein[GFAP]/TUNEL)separately.Results After modeling,the numbers of apoptotic neurons,microvascular endothelial cells and astrocytes in the ischemic cerebral cortex tissue were significantly increased in the model 72 hgroup than in the sham-operation group(P<0.01).Following the treatment,the numbers of the 3 types of apoptotic cells were markedly lower in the eye-acupuncture 72 hgroup than in the model72 hgroup(P<0.01).The expression levels of Bad and Bcl-xL proteins were notably up-regulated in the model 3 h,model 24 h and model 72 hgroups than in the sham operation group(P<0.01).Following eye-acupuncture intervention,modeling induced increase of the Bad expression were obviously reversed in eye-acupuncture 24 hand eye-acupuncture 72 hgroups than those in the2 model groups(P<0.05).And the increase of Bcl-xL expression levels were further increased in the eye-acupuncture groups in comp

关 键 词：脑缺血再灌注 眼针 细胞凋亡 神经血管单元 BAD BCL-XL 

分 类 号：R245.329[医药卫生—针灸推拿学]