针灸调节miR-221-3p/CCR5轴对缺血性脑卒中大鼠神经炎症的影响  

Effects of Acupuncture on Neuroinflammation in Rats with Ischemic Stroke by Regulating miR-221-3p/CCR5 Axis

作　　者：卢菁 王非[1] 张京兰[1] LU Jing;WANG Fei;ZHANG Jinglan(Wuhan Hospital of Traditional Chinese Medicine,Wuhan 430014,Hubei,China)

机构地区：[1]武汉市中医医院,武汉430014

出　　处：《中西医结合心脑血管病杂志》2023年第12期2188-2194,共7页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：武汉市医学科研项目(No.WZ20C41)。

摘　　要：目的:观察针灸调节miR-221-3p/CCR5轴对缺血性脑卒中大鼠神经炎症的影响。方法:采用改良线栓法构建缺血性脑卒中大鼠模型,随机分为模型组、针灸组、miR-221-3p antagomir(拮抗剂)组、miR-221-3p antagomir阴性对照组、针灸+miR-221-3p antagomir组,每组12只;另取12只SD大鼠只分离颈动脉,不结扎、不插线,作为假手术组。大鼠分组以针灸疗法及miR-221-3p antagomir、miR-221-3p antagomir阴性对照处理后,采用Y迷宫实验及跳台实验测定各组大鼠学习记忆能力;对各组大鼠进行神经功能缺损评分并采用苏木精-伊红(HE)染色检测各组大鼠海马神经元形态;采用三苯基氯化四氮唑(TTC)染色检测各组大鼠脑梗死情况;采用酶标仪检测各组大鼠脑组织促炎因子白细胞介素(IL)-17、IL-18及抑炎因子IL-10水平;采用实时荧光定量聚合酶链式反应(RT-PCR)检测各组大鼠脑组织miR-221-3p、CCR5 mRNA表达;采用双荧光素酶实验检测大鼠海马神经元中miR-221-3p对CCR5的靶向调节。结果:与假手术组比较,模型组大鼠海马神经元发生明显病理损伤,新异臂进入次数、新异臂停留时间比、跳台实验潜伏期、脑组织IL-10水平及miR-221-3p mRNA表达降低(P<0.05),跳台实验犯错次数、神经功能缺损评分、脑梗死面积、脑组织IL-17、IL-18水平及CCR5 mRNA表达升高(P<0.05)。与模型组比较,针灸组大鼠海马神经元病理损伤明显减轻,新异臂进入次数、新异臂停留时间比、跳台实验潜伏期、脑组织IL-10水平及miR-221-3p mRNA表达升高(P<0.05),跳台实验犯错次数、神经功能缺损评分、脑梗死面积、脑组织IL-17、IL-18水平及CCR5 mRNA表达降低(P<0.05);与针灸组比较,miR-221-3p antagomir组大鼠海马神经元病理损伤加重,新异臂进入次数、新异臂停留时间比、跳台实验潜伏期、脑组织IL-10水平及miR-221-3p mRNA表达降低(P<0.05),跳台实验犯错次数、神经功能缺损评分、脑梗死面积�Objective:To observe the effects of acupuncture on neuroinflammation in rats with ischemic stroke(IS)by regulating miR-221-3p/C-C chemokine receptor 5(CCR5)axis.Methods:The IS rat model was established by the modified suture method.The rats were randomly divided into model group,acupuncture group,miR-221-3p antagomir(antagonist)group,miR-221-3p antagomir negative control group,and acupuncture+miR-221-3p antagomir group,with 12 rats in each group;another 12 SD rats were separated from carotid artery was separated without ligature or wire was inserted as sham group.After rats were grouped and treated with acupuncture therapy,miR-221-3p antagomir and negative control of miR-221-3p antagomir,the learning and memory ability of rats in each group were measured by Y-maze test and platform jumping test.The neurological deficits of rats in each group were scored and the morphology of hippocampal neurons was detected by hematoxylin-eosin(HE)staining.The cerebral infarcts of each group of rats were detected by triphenyltetrazolium chloride(TTC)staining.The levels of pro-inflammatory factors interleukin(IL)-17,IL-18 and anti-inflammatory factor IL-10 were measured by enzyme marker.The mRNA expression of miR-221-3p and CCR5 in rat brain tissues was measured by rea-l time fluorescence quantitative polymerase chain reaction(RT-PCR),and the target regulation of CCR5 by miR-221-3p in rat hippocampal neurons was detected by dual fluorescein assay.Results:Compared with sham group,hippocampal neurons in the model group had significant pathological injury,and the entry times of new alien arm,residence time ratio of new alien arm,the latency of the platform jumping experiment,the level of IL-10 in brain tissue and mRNA expression of miR-221-3p were decreased(P<0.05).The number of errors,neurological deficit score,cerebral infarction area,the levels of IL-17,IL-18,and mRNA expression of CCR5 in brain tissue were increased(P<0.05).Compared with model group,the pathological injury of hippocampal neurons in the acupuncture group was signi

关 键 词：缺血性脑卒中 针灸 miR-221-3p/C-C趋化因子受体5轴 神经炎症 大鼠 实验研究 

分 类 号：R245[医药卫生—针灸推拿学]