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Application of an atmospheric pressure plasma jet in a rat model of ischaemic stroke:Design,optimisation,and characteristics

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High Voltage 2023年第2期8卷 315-325页

作者： Ye Chen Bingyan Yang Yuqing Liu Lixin Xu Zhongfang Shi Yixiao Liu Ruoyu Han Fang Yuan Jiting Ouyang Xu Yan Kostya(Ken)Ostrikov Department of Pathophysiology Beijing Neurosurgical Institute/Beijing Tiantan HospitalCapital Medical UniversityBeijingChina School of Basic Medical Sciences Capital Medical UniversityBeijingChina School of Physics Beijing Institute of TechnologyBeijingChina School of Chemistry and Physics and Centre for Biomedical Technologies Queensland University of TechnologyBrisbaneQueenslandAustralia

Recent studies have shown the in vitro neuro-protective functions of atmospheric pressure plasma(APP)against multiple pathological injuries during ischaemic stroke(IS).However,whether APP treatment exerts a therapeutic effect on a rat IS model remains unclear.Here,on the basis of needle-to-ring dielectric barrier discharge,an atmospheric pressure plasma jet(APPJ)was designed,with the Helium as the working gas which was driven by a sinusoidal voltage.Then,the treatment conditions were optimised for IS rat model treatment and the characteristics of this APPJ were further diagnosed.Subsequently,the rat IS model was established through 90 min middle cerebral artery occlusion(MCAO),and plasma was intermittently inhaled by rats via the nasal cavity for a 2 min period at 60 min of MCAO process.The therapeutic effects of this plasma jet device were then evaluated using biomedical analyses.According to our results,intermittent APP inhalation in the MCAO rats increased the serum NO content,improved the neurological function,enhanced regional cerebral blood flow,lowered brain infarction,and reduced the cell apoptosis in brain tissues of MCAO rats.Collectively,our data provides a novel potential strategy for IS treatment by using atmospheric-pressure plasma inhalation.

关键词： needle cerebral MCAO treatment

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红景天苷衍生物对苯甲酰红景天苷对MCAO大鼠的神经保护作用

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中国药理学通报 2022年第1期38卷 67-72页

作者：聂婧雯孙春晓黄私迎罗锐赖文芳王颖峥洪桂祝福建中医药大学药学院福建福州350122

目的研究红景天苷衍生物对苯甲酰红景天苷(p-benzoyl salidroside, pOBz)对大脑中动脉闭塞模型(MCAO)大鼠的神经保护作用。方法 (1)30只健康成年♂SD大鼠,随机分为Sham组、MCAO组、MCAO+pOBz低中高剂量组(25、50、100 mg·kg^(-1)),线... 详细信息

目的研究红景天苷衍生物对苯甲酰红景天苷(p-benzoyl salidroside, pOBz)对大脑中动脉闭塞模型(MCAO)大鼠的神经保护作用。方法 (1)30只健康成年♂SD大鼠,随机分为Sham组、MCAO组、MCAO+pOBz低中高剂量组(25、50、100 mg·kg^(-1)),线栓法造模,进行神经功能损伤评分,连续给药2 d。采用核磁共振成像检测MCAO大鼠脑梗死体积。(2)24只健康成年♂SD大鼠,随机分为Sham组、MCAO组、MCAO+pOBz组(50 mg·kg^(-1))和MCAO+Sal组(50 mg·kg^(-1)),线栓法造模,给药1 d。Western blot检测NeuN、EGR1、Bcl-2和Bax蛋白表达。(3)18只健康成年♂SD大鼠,随机分成Sham组、MCAO组和MCAO+pOBz组(50 mg·kg^(-1)),连续给药2 d。免疫荧光染色观察NeuN的表达。结果 pOBz给药2 d可以降低MCAO大鼠的脑梗死体积,改善神经功能损伤,且升高NeuN和EGR1表达程度优于Sal, pOBz对MCAO大鼠脑组织Bcl-2/Bax比值的改善程度与Sal相当。结论 pOBz可改善MCAO大鼠脑梗死体积,具有优于红景天苷的神经保护作用。

关键词：对苯甲酰红景天苷缺血性脑卒中 MCAO 神经保护 NeuN EGR1

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红景天苷预防给药对MCAO大鼠的神经保护作用

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中国药理学通报 2023年第5期39卷 890-896页

作者：黄私迎聂婧雯罗锐周彬彬余正双黄嘉慧赖文芳洪桂祝福建中医药大学药学院福建福州350122

目的研究红景天苷(salidroside,Sal)预防给药对MCAO大鼠的神经保护作用。方法SD成年雄性大鼠52只,随机分:假手术组(Sham)、模型组(MCAO)、红景天苷预给药组(MCAO+Sal)。Sal给药剂量为50 mg·kg^(-1)。连续腹腔注射给药7 d,d 7给药0.5 h... 详细信息

目的研究红景天苷(salidroside,Sal)预防给药对MCAO大鼠的神经保护作用。方法SD成年雄性大鼠52只,随机分:假手术组(Sham)、模型组(MCAO)、红景天苷预给药组(MCAO+Sal)。Sal给药剂量为50 mg·kg^(-1)。连续腹腔注射给药7 d,d 7给药0.5 h后,线栓法制作MCAO模型,24 h后麻醉取材。TTC染色检测大鼠脑梗死体积,TUNEL染色测定神经细胞凋亡数,Western blot测定脑组织中Bax、Bcl-2、cleaved Caspase-3和总Caspase-3蛋白的表达,萘酚AS-D氯乙酸酯酶检测脑组织中性粒细胞的表达,RT-qPCR法分析IL-1β、IL-6、TNF-α的mRNA表达,免疫荧光法分析脑组织中NeuN的表达。结果Sal预给药7 d明显减少了MCAO大鼠脑梗死体积和TUNEL阳性细胞数,促进Bcl-2蛋白的表达,抑制Bax、cleaved Caspase-3蛋白的表达,但对总Caspase-3蛋白无明显作用,减少缺血侧脑组织中性粒细胞的募集,降低炎症因子IL-1β、IL-6、TNF-α的mRNA表达,上调NeuN的表达。结论Sal预给药7 d能明显降低MCAO大鼠脑梗死体积,抑制神经细胞凋亡,减轻炎症,促进NeuN的表达,进而起到神经保护作用。

关键词：红景天苷 MCAO 神经保护凋亡抗炎 NeuN

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大黄对MCAO模型大鼠的神经保护作用

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中国药理学通报 2021年第4期37卷 584-589页

作者：孙春晓黄嘉慧谯丽刘俊杰唐宇恒许阿娟聂婧雯黄私迎罗锐杨泽霖赖文芳洪桂祝福建中医药大学药学院福建福州350122

目的研究大黄提取物对MCAO模型大鼠的神经保护作用,并探讨其作用机制。方法SPF级雄性SD大鼠45只,随机分为sham组、MCAO组、MCAO+大黄组,线栓法制备MCAO模型,大黄提取物给药浓度为200 mg·kg^(-1)·d^(-1),连续给药6 d,尼氏染色法检测尼... 详细信息

目的研究大黄提取物对MCAO模型大鼠的神经保护作用,并探讨其作用机制。方法SPF级雄性SD大鼠45只,随机分为sham组、MCAO组、MCAO+大黄组,线栓法制备MCAO模型,大黄提取物给药浓度为200 mg·kg^(-1)·d^(-1),连续给药6 d,尼氏染色法检测尼氏体的表达,免疫荧光法检测大鼠缺血侧NeuN、NF200的表达,RT-qPCR法检测Egr1、Egr2、Egr4 mRNA表达,Western blot检测NGF及BDNF蛋白的表达。结果与MCAO组相比,大黄提取物作用后,尼氏染色法检测结果表明大黄提取物可以减少MCAO模型大鼠脑梗死面积,免疫荧光检测结果表明大黄提取物可以升高MCAO模型大鼠缺血侧NeuN、NF200的表达;经RT-qPCR检测,大黄提取物能够提高Egr1、Egr2、Egr4 mRNA基因表达;经Western blot检测,大黄提取物能够提高NGF及BDNF蛋白的表达。结论大黄提取物能够改善MCAO模型大鼠缺血侧神经元存活环境,具有神经保护作用。

关键词：大黄提取物 MCAO Egrs NGF BDNF 神经保护

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Salvianolic acid A attenuates ischemia reperfusion induced rat brain damage by protecting the blood brain barrier through MMP-9 inhibition and anti-inflammation

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Chinese Journal of Natural Medicines 2018年第3期16卷 184-193页

作者： ZHANG Wen SONG Jun-Ke ZHANG Xue ZHOU Qi-Meng HE Guo-Rong XU Xiao-Na Yan Rong ZHOU Wen-Xia DU Guan-Hua Beijing Key Laboratory of Drug Target Identification and Drug Screening Institute of Materia Medica Chinese Academy of Medical Sciences & Peking Union Medical College Department of Neuroimmunopharmacology Beijing Institute of Pharmacology and Toxicology State Key Laboratory of Toxicology and Medical Countermeasures

Salvianolic acid A(SAA) is a water-soluble component from the root of Salvia Miltiorrhiza Bge, a traditional Chinese medicine, which has been used for the treatment of cerebrovascular diseases for centuries. The present study aimed to determine the brain protective effects of SAA against cerebral ischemia reperfusion injury in rats, and to figure out whether SAA could protect the blood brain barrier(BBB) through matrix metallopeptidase 9(MMP-9) inhibition. A focal cerebral ischemia reperfusion model was induced by middle cerebral artery occlusion(MCAO) for 1.5-h followed by 24-h reperfusion. SAA was administered intravenously at doses of 5, 10, and 20 mg?kg–1. SAA significantly reduced the infarct volumes and neurological deficit scores. Immunohistochemical analyses showed that SAA treatments could also improve the morphology of neurons in hippocampus CA1 and CA3 regions and increase the number of neurons. Western blotting analyses showed that SAA downregulated the levels of MMP-9 and upregulated the levels of tissue inhibitor of metalloproteinase 1(TIMP-1) to attenuate BBB injury. SAA treatment significantly prevented MMP-9-induced degradation of ZO-1, claudin-5 and occludin proteins. SAA also prevented cerebral NF-κB p65 activation and reduced inflammation response. Our results suggested that SAA could be a promising agent to attenuate cerebral ischemia reperfusion injury through MMP-9 inhibition and anti-inflammation activities.

关键词： Salvianolic 酸 A 局部缺血 MCAO 血大脑障碍 NF-B MMP-9

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Stroke gets in your eyes: stroke-induced retinal ischemia and the potential of stem cell therapy

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Neural Regeneration Research 2020年第6期15卷 1014-1018页

作者： Chase Kingsbury Matt Heyck Brooke Bonsack Jea-Young Lee Cesar V.Borlongan Center of Excellence for Aging and Brain Repair University of South Florida College of Medicine

Stroke persists as a global health and economic crisis,yet only two interventions to reduce stroke-induced brain injury exist.In the clinic,many patients who experience an ischemic stroke often further suffer from retinal ischemia,which can inhibit their ability to make a functional recovery and may diminish their overall quality of life.Despite this,no treatments for retinal ischemia have been developed.In both cases,ischemia-induced mitochondrial dysfunction initiates a cell loss cascade and inhibits endogenous brain repair.Stem cells have the ability to transfer healthy and functional mitochondria not only ischemic neurons,but also to similarly endangered retinal cells,replacing their defective mitochondria and thereby reducing cell death.In this review,we encapsulate and assess the relationship between cerebral and retinal ischemia,recent preclinical advancements made using in vitro and in vivo retinal ischemia models,the role of mitochondrial dysfunction in retinal ischemia pathology,and the therapeutic potential of stem cell-mediated mitochondrial transfer.Furthermore,we discuss the pitfalls in classic rodent functional assessments and the potential advantages of laser Doppler as a metric of stroke progression.The studies evaluated in this review highlight stem cell-derived mitochondrial transfer as a novel therapeutic approach to both retinal ischemia and stroke.Furthermore,we posit the immense correlation between cerebral and retinal ischemia as an underserved area of study,warranting exploration with the aim of these treating injuries together.

关键词： laser Doppler MCAO mesenchymal stem cells mitochondrial network mitochondrial transfer ophthalmic artery optic nerve oxygen-glucose deprivation regenerative medicine retinal ganglion cells visual impairment

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Ginsenoside Rg1 protects against transient focal cerebral ischemic injury and suppresses its systemic metabolic changes in cerabral injury rats

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Acta Pharmaceutica Sinica B 2015年第3期5卷 277-284页

作者： Mingbao Lin Wei Sun Wan Gong Yasi Ding Yuanyan Zhuang Qi Hou State Key Laboratory of Bioactive Substance and Function of Natural Medicines Institute of Materia MedicaChinese Academy of Medical Sciences & Peking Union Medical College Clinical Laboratory Center Beijing Friendship HospitalCapital Medical University College of Basic Medical Science Zhejiang Chinese Medical University

Ginsenoside Rg1(GR),a major bioactive compound of traditional Chinese medicine,such as Panax ginseng or Radix Notoginseng,has been shown to exert neuroprotective effects against ischemic stroke.However,pharmacokinetic studies have suggested that GR could not be efficiently transported through the blood–brain barrier.The mechanism by which GR attenuates cerebral ischemic injury in vivo remains largely unknown.Therefore,this study explored potential neuro-protective effects of GR through its systemic metabolic regulating mechanism by using mass spectrometry–based metabolomic profiling.Rats with middle cerebral artery occlusion(MCAO) were treated with GR intravenously.Their metabolic profiles in serum were measured by gas chromatography coupled with mass spectrometry on 1 and 3 days after MCAO.GR exhibited a potent neuro-protective effect by significantly decreasing the neurological scores and infarct volume in the MCAO rats.Moreover,18 differential metabolites were tentatively identified,all of which appeared to correlate well with these disease indices.Our findings suggested that GR carries a therapeutic potential in stroke possibly through a feed-back mechanism by regulating systematic metabolic mediation.

关键词： Ginsenoside Rg1 MCAO Metabolites Biomarkers

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从AMPK/PGC-1ɑ/NRF-1途径探讨巨刺法改善MCAO大鼠神经损伤的作用机制

从AMPK/PGC-1ɑ/NRF-1途径探讨巨刺法改善MCAO大鼠神经损伤的作用...

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作者：胡梦艺福建中医药大学

学位级别：硕士

目的:本课题拟对大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)模型大鼠侧脑室注射AMPK抑制剂,从AMPK/PGC-1α/NRF-1途径探讨巨刺曲池穴、足三里穴改善MCAO大鼠运动障碍的机制,为临床同类研究提供新靶点及研究基础。方法:将75... 详细信息

目的:本课题拟对大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)模型大鼠侧脑室注射AMPK抑制剂,从AMPK/PGC-1α/NRF-1途径探讨巨刺曲池穴、足三里穴改善MCAO大鼠运动障碍的机制,为临床同类研究提供新靶点及研究基础。方法:将75只280±20g的雄性大鼠,随机等分为假手术、模型组、巨刺组、巨刺+Compound C组(Cmpound C为AMPK抑制剂)和巨刺+PBS组(为说明侧脑室注射的操作不会对实验结果造成明显影响的阴性溶剂对照组)。MCAO模型制备后参照ZeaLonga评分判定大鼠是否模型制备成功,将评分1-3分的大鼠纳入实验,干预前进行磁共振T2WI扫描计算梗死体积,m NSS评分和旷场试验评估神经功能和运动能力。选取巨刺组、巨刺+Compound C组以及巨刺+PBS组大鼠进行左侧肢体的曲池、足三里穴进行电针干预,30 min/次/天,共7天。假手术组和模型组同等条件下模拟抓取。干预结束后,旷场试验测试、Cat Walk步态分析系统检测大鼠的运动功能。功能检测结束后,扫描磁共振T2WI计算梗死体积、免疫荧光法观察神经元表达情况、透射电镜观察神经元内线粒体超微结构和相对数量、通过磷钼酸比色法测定ATP含量、Westernblot法检测AMPK、p-AMPK、PGC-1α及NRF-1蛋白的相对表达。结果:1.神经功能评分结果:干预前,除假手术组外,MCAO模型大鼠m NSS评分升高,基线一致(P>0.05),提示造模成功。干预后,对比模型组,巨刺组m NSS评分较低(P0.05)。2.旷场试验结果:干预前,除假手术外,各组大鼠总运动路程减少(P>0.05),基线一致。干预后,对比模型组,巨刺组的大鼠的总运动路程较长(P0.05)。3.Cat Walk步态分析结果:3.1站立时间:干预后,对各组大鼠站立相时间进行定量分析,以评估病理步态严重程度。与假手术组相比,模型组大鼠四肢的站立时间均有延长(P0.05)。3.2摆动速度:干预后,对各组大鼠摆动速度进行定量分析,以评估病理步态严重程度。与假手术组相比,模型组大鼠四肢的摆动速度均有下降(P0.05)。3.3步长:干预后,对各组大鼠四肢步长进行定量分析,以评估病理步态严重程度。与假手术组相比,模型组大鼠四肢的步长均缩短(P0.05)。4.磁共振扫描计算梗死体积:干预前,除假手术组外,各组MCAO模型大鼠均可见白色梗死区,软件标记梗死区并计算体积后结果显示各组之间无差异(P>0.05)。干预后,结果显示,对比干预前,MCAO模型制备的四组大鼠梗死体积均有下降。对比模型组,巨刺组梗死体积百分比较小(P0.05)。5.免疫荧光染色观察NeuN表达:干预后,与假手术组相比,其余四组的NeuN阳性细胞表达个数较少(P0.05)。6.透射电镜观察线粒体超微结构和数量:干预后,通过透射电子显微镜对神经元内线粒体结构的观察结果显示,与假手术组相比,模型组线粒体嵴结构模糊,形态较圆、长度缩短,可见絮状样变,神经元核组织疏松、水肿,核周围线粒体数量较少;与模型组相比,巨刺组线粒体嵴结构较清晰,形态呈短棒状,线粒体数量较多。对比巨刺组,巨刺+Compound C组线粒体嵴结构断裂、模糊,出现絮状样变,线粒体数量减少;对比巨刺组,巨刺+PBS组的线粒体超微结构和数量差异

关键词：脑卒中巨刺法运动障碍 MCAO AMPK

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哈巴苷对急性脑缺血小鼠神经细胞及caspase非依赖性凋亡通路的影响

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中国药理学通报 2017年第12期33卷 1754-1761页

作者：周炳文楼烨亮钟晓明黄真浙江中医药大学药学院浙江杭州310053

目的研究哈巴苷对急性脑缺血小鼠海马神经细胞、线粒体功能及caspase非依赖性细胞凋亡通路的影响。方法采用大脑中动脉阻塞法(middle cerebral artery occlusion,MCAO)复制小鼠急性脑缺血模型。造模后,各组分别立即尾静脉注射哈巴苷(4、... 详细信息

目的研究哈巴苷对急性脑缺血小鼠海马神经细胞、线粒体功能及caspase非依赖性细胞凋亡通路的影响。方法采用大脑中动脉阻塞法(middle cerebral artery occlusion,MCAO)复制小鼠急性脑缺血模型。造模后,各组分别立即尾静脉注射哈巴苷(4、8、12 mg·kg^(-1))、依达拉奉(3.2 mg·kg^(-1)),模型组及假手术组同法给予等量生理盐水(10 m L·kg^(-1))。造模6 h后,采用流式细胞术检测MCAO小鼠海马神经细胞凋亡率及线粒体膜电位;透射电镜观察MCAO小鼠海马神经细胞线粒体内外膜的清晰度、线粒体嵴的完整性、线粒体基质电子密度的高低变化及线粒体肿胀情况;采用Western blot法检测MCAO小鼠脑组织线粒体中AIF及Endo G的蛋白表达;采用q PCR法检测MCAO小鼠海马组织中AIF及Endo G mRNA的表达。结果造模6 h后,与假手术组相比,模型组小鼠脑组织海马神经细胞凋亡率明显增加(P<0.01);海马神经细胞线粒体膜电位明显降低(P<0.01);神经细胞线粒体超微结构受损严重,线粒体结构松散,可见明显肿胀;脑组织线粒体中AIF及Endo G蛋白表达明显降低,线粒体AIF及Endo G蛋白释放增加(P<0.01);海马组织AIF及Endo G mRNA表达明显增加(P<0.01);与模型组相比,哈巴苷各剂量组小鼠脑组织海马神经细胞凋亡率明显降低(P<0.05,P<0.01);海马神经细胞线粒体膜电位明显升高(P<0.01);神经细胞线粒体超微结构明显改善;哈巴苷8、12 mg·kg^(-1)组小鼠脑组织线粒体中AIF及Endo G蛋白表达明显增加,线粒体AIF及Endo G蛋白释放减少(P<0.05,P<0.01),哈巴苷4 mg·kg^(-1)组小鼠脑组织线粒体中Endo G蛋白表达明显增加,线粒体Endo G蛋白释放明显减少(P<0.05);哈巴苷8、12 mg·kg^(-1)组小鼠海马组织AIF及Endo G mRNA表达明显降低(P<0.05,P<0.01)。结论哈巴苷对急性脑缺血有保护作用,其机制可能与保护大脑神经细胞及线粒体生理活性,抑制caspase非依赖性细胞凋亡通路相关。

关键词：哈巴苷 MCAO 神经细胞凋亡线粒体膜电位线粒体超微结构 caspase非依赖性凋亡通路

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Argon: a novel therapeutic option to treat neuronal ischemia and reperfusion injuries?

引用

Neural Regeneration Research 2015年第7期10卷 1043-1044页

作者： Felix Ulbrich Ulrich Goebel Department of Anesthesiology and Intensive Care Medicine University Medical CenterFreiburg Germany

Neuronal injury and neuroprotection：Ischemia and reperfusion injuries in neuronal cells such as acute ischemic stroke-represent the third leading cause of death in the world.Current therapeutic concepts mainly aim to re-establish cerebral blood flow within a time window of less than 3 hours with the goal of limiting secondary brain injury.

关键词： a novel therapeutic option to treat neuronal ischemia and reperfusion injuries MCAO NDS OGD

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