针刺“百会”“水沟”“大椎”穴对脑缺血再灌注损伤模型大鼠海马区差异circRNAs表达的影响  

Effect of Needling at “Baihui” (GV 20), “Shuigou” (GV 26) and “Dazhui” (GV 14) on Differential CircRNAs Expression in the Hippocampus Cerebral Ischemia Reperfusion Injury Model Rats

作　　者：唐红 汪红娟 江姗姗 吕倩忆 王瑶 谢灿明 李燕平[1] 田浩梅[1] 陈楚淘[1] TANG Hong;WANG Hongjuan;JIANG Shanshan;LYU Qianyi;WANG Yao;XIE Canming;LI Yanping;TIAN Haomei;CHEN Chutao(School of Acupuneture.Moxibustion and Tuina,Hunan University of Chinese Medieine,Changsha,410208;Chengdu Second People's Hospital.Sichuan Province)

机构地区：[1]湖南中医药大学针灸推拿与康复学院,湖南省长沙市410208 [2]四川省成都市第二人民医院

出　　处：《中医杂志》2023年第5期512-517,共6页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81874508,82274662);湖南省自然科学基金(2021JJ30490);湖南省研究生科研创新课题(CX20220799);长沙市科技局自然科学基金(kq2014094);湖南中医药大学研究生创新课题(2022CX97,2022CX103)。

摘　　要：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30 min,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|log2 FC|≥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 investigate the possible mechanism of needling at Dazhui(GV 14), Baihui(GV 20) and Shuigou(GV 26) in the treatment of cerebral ischemia reperfusion injury(CIRI) from the perspective of hippocampus gene expression. Methods Thirty-nine SD rats were randomly divided into sham operation group, model group, and acupunture group with 13 rats in each group. The CIRI model in rats was established by suture method in the model group and acupuncture group. After modelling, the sham operation group and model group were only bundled without needling, while the acupuncture group was bundled and treated by needling at Dazhui, Baihui and Shuigou with 30 min of needle retention and performing twirling method every 15 min, using even supplementation and drainage method, once every 12 h, 7 times altogether. The neurological deficit scores were evaluated before and after treatment. TTC staining was used to detect the status of cerebral infarction after treatment. The circRNAs microarry chip was used to analyze differential circRNAs expression in the hippocampus tissue of the ischemic side of CIRI rats, and qRT-PCR was used to verify it. Results Before treatment, the neurological deficit score of model group and acupuncture group were lower than that of the sham operation group(P<0. 01);after treatment, the neurological deficit score of acupuncture group significantly increased, and was higher than that of the model group(P<0. 05).Compared to that of the model group, the cerebral infarction ratio in the acupuncture group was reduced(P<0. 05).CircRNA microarray analysis showed that 215 circRNAs were differentially expressed in the model group/sham operation group, including 59 up-regulated and 156 down-regulated ones. Five circRNAs were differentially expressed in the acupuncture group/sham operation group, of which one was up-regulated and four were down-regulated. Compared with the model group, there were 2 up-regulated circRNAs in acupuncture group. There were two differential genes coexpressed in the model group/sham

关 键 词：脑缺血再灌注损伤 针刺 神经功能缺损 脑梗死面积 circRNAs 海马区 

分 类 号：R245[医药卫生—针灸推拿学]