针刺干预脑出血的铁死亡应激机制  

Protective Mechanism of Acupuncture on Brain of Spontaneous Hypertension Rats by Ferroptosis Pathway

作　　者：高莹[1] 杨建[1] GAO Ying;YANG Jian(First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion,Tianjin 300183,China)

机构地区：[1]天津中医药大学第一附属医院,国家中医针灸临床医学研究中心,天津300183

出　　处：《辽宁中医杂志》2024年第8期190-193,I0006,共5页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金青年项目(82104998)。

摘　　要：目的脑出血后脑组织发生铁死亡应激,针刺作为治疗脑出血的有效手段,其治疗脑出血是否与铁死亡应激有关,目前证据不足。该研究以铁死亡应激为切入点,以醒脑开窍针刺法为干预措施,观察针刺对脑出血大鼠神经功能的影响,探讨针刺干预脑出血的铁死亡应激机制。方法成年雄性SD大鼠(240±10)g,采用自体血注入法制备脑出血模型,30只造模成功的大鼠随机分为模型组、针刺组、DFO(铁死亡抑制剂)组。10只同体质量的雄性SD大鼠为对照组,接受与模型组相同的操作,注等量0.9%氯化钠溶液,不注血。针刺组选取人中、内关、三阴交穴进行针刺,每日1次,一共3日。DFO组腹腔注射50 mg/kg的DFO,每日1次,共3日。采用Ludmila Belayev 12分法对其神经功能进行评分,透射电镜观察各组大鼠脑组织形态学改变,流式细胞术检测ROS水平,普鲁士蓝染色法检测铁沉积,分光光度计测定丙二醛(MDA)含量,免疫荧光检测谷胱甘肽过氧化物酶4(Glutathione peroxidase,GPX4)表达。结果针刺可提高脑出血大鼠神经功能评分,减轻神经缺损症状。透射电镜示模型组神经元内细胞器降解,线粒体数量减少,线粒体嵴排列紊乱,针刺组较模型组线粒体数量及形态改变减轻,DFO组线粒体结构较完整。脑出血大鼠铁沉积增多,活性氧(reactive oxygen species,ROS)、MDA水平升高,GPX4表达下降,针刺可减少铁沉积,降低ROS、MDA水平,促进GPX4表达,差异有统计学意义。结论脑出血后脑组织发生铁死亡应激,针刺可抑制铁死亡应激减轻神经缺损症状。Objective Ferroptosis stress occurs in brain tissue after cerebral hemorrhage.Acupuncture is an effective method for the treatment of cerebral hemorrhage,but whether it is related to ferroptosis stress is insufficient.Taking ferroptosis stress as the entry point and restoring consciousness and opening orifices acupuncture as the intervention measure,this study observed the effect of acupuncture on neurological function in intracerebral hemorrhage(ICH)rats,so as to explore the mechanism of acupuncture on ferroptosis stress in ICH.Methods ICH model was established in adult male SD rats[(240±10)g]by injection of autologous blood.Thirty successfully modeled rats were randomly divided into a model group,an acupuncture group and a iron death inhibitor(DFO)group.Ten male SD rats with the same body weight were used as the control group,and received the same operation as the model group,and the same volume of normal saline was injected without blood injection.In the acupuncture group,acupuncture was applied to Zhongren(CV4),Neiguan(PC6)and Sanyinjiao(SP6),once a day for 3 days.In the DFO group,DFO(50 mg/kg)was injected intraperitoneally once a day for 3 days.Ludmila Belayev′s 12-point method was used to score the neurological function of the rats.Transmission electron microscopy was used to observe the morphological changes of brain tissue.Flow cytometry was used to detect the level of reactive oxygen species(ROS),Prussian blue staining was used to detect the iron deposition,spectrophotometer was used to measure the content of malondialdehyde(MDA),and immunofluorescence was used to detect the expression of glutathione peroxidase 4(GPX4).Results Acupuncture could improve the neurological function score and alleviate the symptoms of neurological deficit in rats with ICH.Transmission electron microscopy showed that the organelles in the neurons were degraded,the number of mitochondria was reduced,and the arrangement of mitochondrial cristae was disordered in the model group.Compared with those of the model group,the numbe

关 键 词：针刺 脑出血 铁死亡应激 醒脑开窍针刺法 

分 类 号：R246.1[医药卫生—针灸推拿学]