电针“水沟”穴对脑缺血大鼠缺血脑组织血管新生相关因子表达的影响  

Effects of electroacupuncture on morphology of neovascularization and expression of angiogenesis-related factors in ischemic brain tissue of cerebral ischemia rats

作　　者：贾蓝羽 杜元灏[2] 李晶[2] 庞博 徐梦瑶 JIA Lan-yu;DU Yuan-hao;LI Jing;PANG Bo;XU Meng-yao(The Affiliated Hospital of Tianjin Academy of Traditional Chinese Medicine,Tianjin 300120,China;Section of Acu-moxibustion,First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300193;Graduate School of Tianjin University of Traditional Chinese Medicine,Tianjin300193)

机构地区：[1]天津市中医药研究院附属医院,天津300120 [2]天津中医药大学第一附属医院针灸部,天津300193 [3]天津中医药大学研究生院,天津300193

出　　处：《针刺研究》2019年第10期715-721,共7页Acupuncture Research

基　　金：国家自然科学基金项目(No.81473765)

摘　　要：目的:观察脑梗死后不同时间点缺血区脑组织中新生血管形态及血管新生相关因子的变化规律,探讨电针"水沟"穴对脑梗死后血管新生的干预效应。方法:雄性Wistar大鼠随机分为假手术组、模型组、电针组各90只,空白组10只。采用大脑中动脉线栓法制备大脑中动脉栓塞(MCAO)模型,并分为缺血1、3、6、9、12、24 h及3、7、12 d共9个时间点组。电针组以15 Hz、2 mA的连续波电针"水沟"穴20 min,1~24 h电针组于造模后即刻给予电针干预,相应时间点取材,3~12 d电针组每日干预1次,末次治疗后取材。采用免疫荧光双标记法观察缺血区脑组织中新生血管形态和数目,采用实时荧光定量PCR和Western blot法测定梗死脑组织中碱性成纤维细胞生长因子(bFGF)、血管生长素(Ang)-1和2、血小板源性生长因子b(PDGF-b)的表达水平。结果:空白组、假手术组未出现CD31和Ki67双阳性细胞;模型组24 h开始出现CD31和Ki67双阳性细胞,随缺血时间延长双阳性细胞逐渐增多,3 d时达到峰值,7 d时下降,12 d时无双阳性细胞;电针组12 h时CD31和Ki67双阳性细胞开始出现,3 d时达峰值后逐渐下降,12 d时仍有少量双阳性细胞。假手术组与空白组bFGF、Ang-1、Ang-2、PDGF-b mRNA及蛋白表达比较差异无统计学意义(P>0.05)。模型组bFGF mRNA表达在缺血后9 h^12 d时、蛋白表达在24 h时高于假手术组(P<0.01,P<0.05);Ang-1mRNA表达在缺血后12 h^12 d、蛋白表达在6 h^12 d时高于假手术组(P<0.05,P<0.01);Ang-2 mRNA及蛋白表达在缺血后1 h^12 d时高于假手术组(P<0.01,P<0.05);PDGF-b mRNA在1、6、9、24 h及3~12 d时,蛋白表达在1 h^7 d时高于假手术组(P<0.05,P<0.01)。电针组bFGF mRNA在24 h^12 d、蛋白表达在3~12 d时高于模型组(P<0.05,P<0.01);Ang-1 mRNA及蛋白表达在3~12 d时高于模型组(P<0.05,P<0.01);Ang-2 mRNA表达在3~24 h、蛋白表达在3~12 h时高于模型组(P<0.05,P<0.01);PDGF-b mRNA在3 h、6 h、3~12 d时,蛋白表达�Objective To observe the change of neovascular morphology and angiogenesis related factors in the ischemic cerebral area after cerebral infarction and the intervention effect of electroacupuncture(EA).Methods Wistar rats were randomly divided into model group(n=90),EA group(n=90),sham operation group(n=90)and control group(n=10).The first three groups were further divided into 1 h,3 h,6 h,9 h,12 h,24 h,3 d,7 d and 12 d subgroups(n=10 in each subgroup).The cerebral infarction model was established by middle cerebral artery occlusion(MCAO).EA(15 Hz,2 mA)was applied to"Shuigou"(GV26)for 20 min in the EA group.The 1,3,6,9,12,24 h subgroups were treated immediately after modeling,the 3,7,12 d subgroups were treated once daily for 3,7 or 12 days.The neovascular endothelial cells were displayed by immunofluorescence double labeling staining.Quantitive real-time PCR and Western blot were applied to detect the changes of basic fibroblast growth factor(bFGF),angiogenin(Ang)-1,2,platelet-derived growth factor b(PDGF-b)in ischemic brain tissue,respectively.Results After modeling,CD31 and Ki67 positive cells were first observed at 24 h in the model group,and reached the peak at 3 d,decreased at 7 d.While in the EA group,the CD31 and Ki67 positive cells were first observed at 12 h,and reached the peak at 3 d,and gradually decreased until 12 d.Compared with the control group,the mRNA expressions of bFGF at 9 h-12 h,Ang-1 at 12 h-12 d,Ang-2 at 1 h-12 d and PDGF-b at 1 h,6 h,9 h,24 h-12 d were increased in the model group(P<0.01,P<0.05).After EA,the mRNA expressions of bFGF at 24 h-12 d,Ang-1 at 3 d-12 d,Ang-2 at 3 h-24 h and PDGF-b at 3 h,6 h,3 d-12 d were increased(P<0.05,P<0.01).In comparison with the control group,the proteins of bFGF at 24 h,Ang-1 at6 h-12 d,Ang-2 at 1 h-12 d and PDGF-b at 1 h-7 d were increased in the model group(P<0.05,P<0.01).After EA,the proteins of bFGF at 3 d-12 d,Ang-1 at 3 d-12 d,Ang-2 at 3 h-12 h and PDGF-b at 6 h,3 d-12 d were increased compared with the model group(P<0.05,P<0.01).Conclusion EA can

关 键 词：大脑中动脉栓塞 血管新生 电针 水沟穴 

分 类 号：R245.97[医药卫生—针灸推拿学]