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眼针联合尾灸对脑卒中后认知功能障碍大鼠的影响与机制研究OA北大核心CSTPCD

Effect and mechanism of ophthalmic acupuncture combined with tail moxibustion in post-stroke cognitive impairment rats

中文摘要英文摘要

目的 探讨眼针联合尾灸对脑卒中后认知功能障碍大鼠行为学、海马的影响和可能起效机制.方法 75只雄性SD大鼠采用随机数字表法分为假手术组(15只)和造模组(60只).造模组采用改良线栓法复制大脑中动脉闭塞模型,假手术组仅暴露右侧颈动脉.造模后通过Morris水迷宫实验筛选脑卒中后认知功能障碍大鼠48只,采用随机数字表法分为模型组、眼针组、尾灸组、眼针+尾灸组,每组12只.假手术组、模型组仅捆绑,不予任何干预;眼针组取双侧"肝区""肾区""心区""脾区",留针30 min,每日1次;尾灸组艾灸"长强"和尾尖之间区域,温和灸20 min,每日1次;眼针+尾灸组同时给予上述眼针、尾灸干预.干预7 d后进行大鼠行为学检测;苏木素-伊红染色观察海马病理学变化;比色法检测海马中丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性;蛋白质印迹法检测大鼠海马Kelch样环氧氯丙烷相关蛋白1(KEAP1)、磷酸甘油酸变位酶5(PGAM5)、线粒体相关凋亡诱导因子1(AIFM1)、核转录因子E相关因子2(Nrf2)、血红素加氧酶-1(HO-1)蛋白表达;实时荧光定量PCR法检测海马KEAP1、PGAM5、AIFM1、Nrf2、HO-1 mRNA表达.结果 与假手术组比较,模型组大鼠逃逸潜伏期延长,穿越平台次数减少(均P<0.05);大鼠海马CA1区神经元数量明显减少,排列紊乱,形态不规则,可见坏死的神经元;海马中SOD活性降低,MDA含量升高(均P<0.05);KEAP1、PGAM5、AIFM1蛋白及mRNA表达升高,Nrf2、HO-1蛋白及mRNA表达降低(均P<0.05).与模型组比较,眼针+尾灸组大鼠逃逸潜伏期缩短,穿越平台次数增加(均P<0.05);大鼠海马CA1区神经元丢失明显减少,细胞形态较饱满;海马中SOD活性升高,MDA含量降低(均P<0.05);KEAP1、PGAM5、AIFM1蛋白及mRNA表达降低,Nrf2、HO-1蛋白及mRNA表达升高(均P<0.05).结论 眼针联合尾灸可治疗脑卒中后认知功能障碍大鼠,其机制可能是通过减轻海马氧化应激损伤和氧死亡,从而改善海马神经元损伤程度,提高脑卒中后大鼠的认知能力.

Objective To explore the effects and possible mechanisms of ophthalmic acupuncture combined with tail moxibustion on the behavior and hippocampus in rats with post-stroke cognitive impairment.Methods Seventy-five male SD rats were randomly divided into the sham operation group (15 rats) and the modeling group (60 rats) using the random number table method. A modified suture-occluded method was used to replicate the occlusion model of the middle cerebral artery in the modeling group,whereas only the right carotid artery was exposed in the sham operation group. After modeling,48 rats with post-stroke cognitive impairment were selected using the Morris water maze experiment,and were randomly divided into the model group,the ophthalmic acupuncture group,the tail moxibustion group and the ophthalmic acupuncture+tail moxibustion group using the random number table method,with 12 rats per group. The sham operation group and the model group were bound with no intervention;the ophthalmic acupuncture group was needled once a day in the bilateral "liver area","kidney area","heart area",and "spleen area",leaving the needle for 30 min;the tail moxibustion group was given mild-warm moxibustion on the area between "Changqiang" (GV1) and the tip of the tail for 20 min,once a day;the ophthalmic acupuncture+tail moxibustion group was given the above-mentioned ophthalmic acupuncture and tail moxibustion interventions simultaneously. After 7 days of intervention,the behavior of the rats was detected. Hematoxylin and eosin staining was used to observe the pathological changes in the hippocampus;the malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in the hippocampus were detected by colorimetric method;Western blotting was used to detect the protein expressions of Kelch-like ECH associated protein 1 (KEAP1),phosphoglycerate mutase 5 (PGAM5),apoptosis-inducing factor mitochondria associated 1 (AIFM1),nuclear factor erythroid 2-related factor 2 (Nrf2),and heme oxygenase-1 (HO-1) in the hippocampus of rats;and real-time fluorogenic quantitative PCR was used to determine the mRNA expressions of KEAP1,PGAM5,AIFM1,Nrf2,and HO-1 in the hippocampus. Results Compared with the sham operation group,the escape latency in the model group was prolonged,and the crossing platform number was decreased (P<0.05);the number of neurons in the hippocampal CA1 area was significantly decreased,with a disordered arrangement and irregular morphology,and necrotic neurons were observed;the SOD activity in the hippocampus was decreased,while the MDA content was increased (P<0.05);the protein and mRNA expressions of KEAP1,PGAM5,and AIFM1 were increased,while the protein and mRNA expressions of Nrf2 and HO-1 were decreased (P<0.05). Compared with the model group,the escape latency of rats in the ophthalmic acupuncture+tail moxibustion group was shortened,and the crossing platform number was increased (P<0.05);the loss of neurons in the hippocampal CA1 area of the rat was significantly reduced,and the cell morphology was more plump;SOD activity in the hippocampus was increased,and MDA content was decreased (P<0.05);and the protein and mRNA expressions of KEAP1,PGAM5,and AIFM1 were decreased,while the protein and mRNA expressions of Nrf2 and HO-1 were increased (P<0.05).Conclusion The combination of ophthalmic acupuncture and tail moxibustion can be used to treat rats with post-stroke cognitive impairment,and its mechanism may be related to alleviating oxidative stress damage and oxeiptosis in the hippocampus,thereby improving the degree of hippocampal neuronal damage and enhancing the cognitive ability of rats after stroke.

唐鑫怡;张威;周鸿飞

辽宁中医药大学针灸推拿学院 沈阳 110847辽宁中医药大学附属医院

中医学

眼针尾灸脑卒中后认知功能障碍氧化应激氧死亡大鼠

ophthalmic acupuncturetail moxibustionpost-stroke cognitive impairmentoxidative stressoxeiptosisrats

《北京中医药大学学报》 2024 (009)

1312-1321 / 10

国家自然科学基金项目(No.81703993);辽宁省应用基础研究计划项目(No.2023JH2/10130051)National Natural Science Foundation of China(No.81703993)

10.3969/j.issn.1006-2157.2024.09.016

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