“智三针”通过自噬途径抑制卒中后抑郁大鼠海马神经元炎性坏死的机制研究  

Mechanism of“Zhi san zhen”inhibited hippocampal neuronal inflammatory necrosis through regulating autophagy in rats with ischemic post⁃stroke depression

作　　者：武莉华 高静 王孟雨 傅文[4] 史册 赖名殷 WU Lihua;GAO Jing;WANG Mengyu;FU Wen;SHI Ce;LAI Mingyin(Seventh Clinical Medical College of Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Rehabilitation Center,The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;College of Rehabilitation Medicine,Henan University of Chinese Medicine,Zhengzhou 450046,China;Rheumatology Department,The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;The Third Clinical Medical College,Henan University of Traditional Chinese Medicine,Zhengzhou 450003,China;School of Traditional Chinese Medicine,Hainan Medical University,Haikou 571199,China)

机构地区：[1]广州中医药大学第七临床医学院,广东广州510006 [2]河南中医药大学第一附属医院康复中心,河南郑州450000 [3]河南中医药大学康复医学院,河南郑州450046 [4]河南中医药大学第一附属医院风湿科,河南郑州450000 [5]河南中医药大学第三临床医学院,河南郑州450000 [6]海南医科大学中医学院,海南海口571199

出　　处：《海南医学院学报》2024年第13期961-969,共9页Journal of Hainan Medical University

基　　金：国家自然科学基金区域科学基金项目(82260968);海南省自然科学高层次人才基金项目(821RC744)。

摘　　要：目的:观察“智三针”对卒中后抑郁(PSD)模型大鼠海马神经元自噬、炎症水平的影响。方法:63只雄性SD大鼠随机分为7组,每组9只。除假手术组外,其余各组先构建大脑中动脉阻塞再灌注(MCAO/R)模型,术后第4天起采用孤养联合CUMS 21 d构建缺血性PSD模型。造模24 h后,予针刺组“智三针”针刺干预,帕罗西汀组盐酸帕罗西汀(10 mg/kg)灌胃给药,假针刺组非穴不透皮假针干预,3‑MA组自噬抑制剂3‑MA(2.5 mg/kg)腹腔注射,3‑MA+针刺组3‑MA注射后针刺干预。1次/d,共14 d。神经功能缺损评分和糖水偏好率评估大鼠行为学,Western blot检测海马组织Beclin‑1、BNIP3、LC3、NLRP3、NF‑κB、IL‑1β、IL‑18蛋白表达,HE染色观察海马组织病理形态,透射电镜观察海马神经元超微结构和自噬小体。结果:造模后,与假手术组相比,模型组神经功能缺损加重(P<0.001),糖水偏好上升(P<0.01),海马Beclin‑1、BNIP3、LC3、NLRP3、NF‑κB、IL‑1β、IL‑18表达均升高(P<0.05)。干预后,与模型组相比,针刺组神经功能缺损改善(P<0.001),糖水偏好上升(P<0.001),海马Beclin‑1、BNIP3、LC3表达升高(P<0.001),NLRP3、NF‑κB、IL‑1β、IL‑18表达降低(P<0.01);帕罗西汀组神经功能缺损无显著差异(P>0.05),糖水偏好率上升(P<0.001),Beclin‑1、BNIP3、LC3表达升高(P<0.01),NLRP3、NF‑κB、IL‑1β、IL‑18表达降低(P<0.01);假手术组及假针刺组各项指标均无显著差异(P>0.05);3‑MA组神经功能缺损加重(P<0.05),糖水偏好下降(P<0.05),海马Beclin‑1、BNIP3、LC3表达降低(P<0.01),NLRP3、NF‑κB、IL‑1β、IL‑18表达均显著升高(P<0.01);与3‑MA组相比,3‑MA+针刺组神经功能缺损改善(P<0.001),糖水偏好上升(P<0.001),Beclin‑1、BNIP3、LC3表达升高(P<0.001),NLRP3、NF‑κB、IL‑1β、IL‑18表达下降(P<0.01);HE染色及透射电镜观察可见,假手术组海马神经元结构完整,未见自噬小体;PSD模型组和假针刺组海�Objective:To observe the effects of"Zhi san zhen"on on autophagy and inflammation levels in hippocampal neurons of rats with post‑stroke depression(PSD).Methods:63 male SD rats were randomly divided into seven groups,nine in each group.The middle cerebral artery occlusion reperfusion(MCAO/R)model was constructed first in all groups except the sham‑operated group,and the ischaemic PSD model was constructed using solitary support combined with CUMS for 21 d postoperatively onwards from the 4th postoperative day.After the model was successfully constructed,the acupuncture group was treated with acupuncture at"Zhi san zhen"acupoints.The drug group received paroxetine hydrochloride(10 mg/kg)by gavage.The sham acupuncture group received non‑acupoint transdermal sham needle intervention.The 3‑MA group received the autophagy inhibitor 3‑MA(2.5 mg/kg)by intraperitoneal injection.The acupuncture+3‑MA group received 3‑MA injection followed by acupuncture intervention.All interventions were given once daily for 14 consecutive days.Neurological function score and sucrose preference test of rats were detected after intervention;Western blot was used to detect the protein expression levels of Beclin‑1,LC3,BNIP3,NF‑κB,NLRP3,IL‑1β,IL‑18 in hippocampal tissue.HE staining was used to observe the histopathological morphology of the hippocampus,and observe the hippocampal neurons and autophagic vesicles by transmission electron microscopy.Results:The model group had elevated neurological function scores(P<0.001),decreased sucrose preference(P<0.01),and elevated hippocampal expression of Beclin‑1,BNIP3,LC3,NLRP3,NF‑κB,IL‑1β,IL‑18(P<0.05)compared with the sham‑operated group;Compared with the model group,the acupuncture group had lower neurological function scores(P<0.001),higher sucrose preference(P<0.001),higher hippocampal expression of Beclin‑1,BNIP3,LC3(P<0.001),and lower expression of NLRP3,NF‑κB,IL‑1β,IL‑18(P<0.01);the paroxetine group had no significant difference in neurological fu

关 键 词：缺血性卒中后抑郁 智三针 细胞自噬 炎性坏死 

分 类 号：R245[医药卫生—针灸推拿学]