摘要
目的观察醒脑开窍针刺法对脑缺血再灌注损伤大鼠神经功能和脑组织病理形态学的影响,检测脑缺血再灌注组织内谷胱甘肽(glutathione,GSH)、丙二醛(malondialdehyde,MDA)和谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)、转铁蛋白受体(transferrin receptor,TfR)和膜铁转运蛋白1(ferroportin 1,Fpn1)含量变化,从铁死亡途径探讨针刺对脑缺血再灌注大鼠的脑保护机制。方法25只清洁级雄性Wistar大鼠随机分为假手术组5只、模型组10只、电针组10只。后两组分别再按针刺干预时间分为24 h、72 h两个亚组,每组5只。采用改进的Zea Longa线栓法制作大脑中动脉缺血再灌注模型。电针组予针刺内关、水沟、三阴交,留针期间患侧内关、三阴交穴连接神经穴位刺激仪。采用Zausinger六分法评定神经功能缺损程度,尼氏染色法观察脑组织病理形态改变,酶联免疫吸附测定法对比各组大鼠GSH、MDA、TfR、Fpn1、GPX4的含量。结果与假手术组比较,造模后模型组及电针组神经功能评分均降低(P<0.01)。与模型组比较,电针干预24 h后神经功能评分升高,72 h后评分进一步升高(P<0.01)。造模24 h后,模型组及电针组尼氏小体数量减少,颜色变浅;72 h后模型组尼氏小体更少,电针组尼氏小体数量有一定的恢复。与假手术组比较,模型组MDA、TfR、Fpn1升高、GSH及GPX4降低(P<0.01)。与模型组比较,电针干预24 h及72 h时后MDA、TfR降低、GSH、GPX4及Fpn1升高(P<0.01,P<0.05)。与针刺24 h对比,针刺72 h后MDA、TfR降低更明显,GSH及Fpn1升高更明显(P<0.01,P<0.05)。结论铁死亡是脑缺血再灌注损伤中细胞死亡的重要方式之一,醒脑开窍针刺法可调节脑缺血再灌注大鼠脑组织铁代谢,提高细胞抗氧化能力,抑制神经细胞铁死亡,其脑保护作用与铁死亡途径密切相关。
Objective To observe the effect of“restoring consciousness and opening orifices”acupuncture on the cerebral ischemia reperfusion injury rats,detect the differences in the expressions of glutathione(GSH)content,malondialdehyde(MDA),transferrin receptor(TfR),ferroportin1(Fpn1)and glutathione peroxidase 4(GPX4)in the areas of cerebral ischemia reperfusion injury and explore the protective mechanism of acupuncture on the cerebral ischemia reperfusion injury rats from the way of ferroptosis.Methods In this study,25 specific pathogen free grade male Wistar rats were used as the research subjects.There were five rats in sham operation group,ten rats in model group and ten rats in the electroacupuncture(EA)group.According to the intervention time(24 h and 72 h),the model group and EA group were divided into two subgroups respectively,5 rats in each group.The cerebral ischemia reperfusion model was established by Zea Longa′s suture method in the model group and the EA group.EA was performed at“Neiguan”(PC6),“Shuigou”(GV26)and“Sanyinjiao”(SP6)in the EA group.EA was connected at the affected“Neiguan”(PC6)and“Sanyinjiao”(SP6).After treatment,neurological function was assessed by Zausinger′s neurologic assessment scale,Nissl staining was used to observe the pathological changes in brain tissue.ELISA method was used to detect the contents of GSH,GPX4,MDA,TfR and Fpn1.Results Compared with those of the sham operation group,the neurological function scores of model group and EA group were decreased(P<0.01).Compared with those of the model group,the neurological function scores of EA group were increased after 24 h EA intervention and were more significantly increased after 72 h EA intervention(P<0.01).After 24 h intervention,the numbers of nissler bodies in the model group and EA group were both decreased and the color of nissler bodies both became lighter.After 72 h intervention,the numbers of nissler bodies in the model group were reduced more,but the numbers of nissler bodies in EA group were increased.Compared with those of the sham operation group,the contents of GSH and GPX4 in focal brain tissue were both decreased in the model group,the contents of MDA,TfR and Fpn1 were all increased in the model group(P<0.01).Comparing with the model group,after 24 h and 72 h EA intervention,the contents of MDA and TfR in the EA group were significantly decreased,the contents of GSH,GPX4 and Fpn1 in the EA group were significantly increased(P<0.01,P<0.05).The reduction of the contents of MDA and TfR in the EA group were more significantly,and the contents of GSH and Fpn1 in the EA group were increased more significantly after 72 h intervention(P<0.01,P<0.05).Conclusions:Ferroptosis is one of the important ways of cell death in cerebral ischemia reperfusion injury.The mechanism of“restoring consciousness and opening orifices”acupuncture may be related to regulate the iron metabolism and improve the cell antioxidant ability,the protective effect of acupuncture on cerebral ischemia reperfusion injury rats is closely related to the way of ferroptosis.
作者
王琪
李澎
许军峰
WANG Qi;LI Peng;XU Junfeng(National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion,Tianjin 300193,China;First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300193,China)
出处
《辽宁中医杂志》
CAS
北大核心
2024年第6期196-200,I0011,共6页
Liaoning Journal of Traditional Chinese Medicine
基金
国家自然科学基金项目(82104995)。
