PEP-1介导铜锌超氧化物歧化酶对帕金森病小鼠氧化应激损伤的保护作用

目的 观察PEP-1-铜、锌超氧化物歧化酶(PEP-1-SOD1)对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)所致帕金森病小鼠行为学和氧化应激损伤的影响,探讨其可能的神经保护机制.方法 36只C57/BL小鼠随机分为正常组、模型组和PEP-1-SOD1组.模型组给MPTP腹腔注射,连续5 d;PEP-1-SOD1组连续腹腔注射MPTP 5 d前给予PEP-1-SOD1腹腔注射3 d.通过爬杆实验观察小鼠行为学变化.取小鼠纹状体通过黄嘌呤氧化酶法检测铜/锌超氧化物歧化酶(SOD1)和谷胱甘肽过氧化物酶(GSH-Px)活性,通过TBA法检测MDA含量.结果 模型组小鼠爬杆能力明显下降,PEP-1-SOD1组处理的小鼠行为学异常明显改善(P＜0.01).模型组小鼠纹状体SOD1和GSH-Px活性明显降低,MDA含量显著升高(P＜0.01).同模型组比较,PEP-1-SOD1组小鼠纹状体SOD1和GSH-Px活性显著增高,MDA含量明显减少(P＜0.01).结论 PEP-1-SOD1具有神经保护作用,其机制可能与减轻氧化应激损伤、增强抗自由基损伤能力有关.     

铜及铜锌超氧化物歧化酶对神经保护作用的研究进展

铜参与神经系统功能的调节,有神经元保护及髓鞘修复功能,铜锌超氧化物岐化酶(copper zinc superoxide dis-mutase,SOD1)是一种含铜酶,对脊髓及脑损伤有神经保护作用。本文探讨铜及SOD1的神经保护作用及神经损伤修复及临床治疗的新途径。     

利拉鲁肽对脑梗死小鼠保护作用的研究

目的探讨利拉鲁肽对实验性脑梗死小鼠的保护作用及分子机制。方法 C57BL/6小鼠随机分为假手术组、对照组和利拉鲁肽组。烧灼法建立永久、局灶性右侧皮质梗死模型(dMCAO),假手术组仅分离血管。利拉鲁肽组于dMCAO术后连续3d分别给予腹腔注射利拉鲁肽(200μg·kg-1·d-1),对照组腹腔注射等体积生理盐水。TTC染色方法计算脑梗死体积,Rota-rod方法评价肢体功能。术后72h时采用Western blot测定三个实验组p-AMPK/AMPK和NF-κB蛋白水平。结果利拉鲁肽明显延长dMCAO小鼠的Rota-rod时间(52.22±4.588 s),并缩小梗死体积(0.0791±0.0025),上调p-AMPK蛋白水平和p-AMPK/AMPK比值,抑制NF-κB表达。结论利拉鲁肽减轻dMCAO小鼠炎症反应和缺血性脑损伤,激活AMPK、抑制NF-κB是利拉鲁肽保护缺血性脑损伤的分子机制。     

1周内TIA对老年人首发脑梗死的保护作用

目的　探讨 1周内TIA对老年人首发脑梗死有无保护作用。方法　将首发脑梗死老年患者分为观察组 (病前 1周内有TIA)和对照组 (病前 1周内无TIA) ,观察意识障碍发生率、病死率、神经功能有缺损评分及脑血流量。结果　观察组意识障碍发生率、神经功能缺损评分低于对照组 ,而脑血流量高于对照组 ,且治疗一月后疗效好 ,但病死率无明显差别。结论　 1周内TIA对老年人首发脑梗死有保护作用。     

SOD1基因突变子对AD转基因小鼠β-淀粉样蛋白表达影响的实验研究

目的探讨Cu/Zn SOD1基因表达对Aβ产生的影响.方法建立APP-C100和G93A SOD1基因共表达的转双基因小鼠,应用免疫细胞化学方法测定不同鼠龄转基因鼠海马神经元Aβ表达,Westem blot定量测定转基因鼠脑SCD1、APP、APP-C100和Aβ表达水平.结果低龄转双基因鼠与转单基因鼠之间Aβ水平无差异,与年龄匹配的转单基因鼠比较,高龄转双基因鼠脑Aβ水平升高,经统计处理差异无显著性意义(P＞O.05).但高龄转双基因鼠Aβ水平比低龄转双基因鼠明显升高(P＜0.05).结论转双基因鼠脑APP-C100和G93A SOD1基因共表达虽然未导致老年斑形成,但引起Aβ含量增加,推测其机制可能与SOD1基因突变子引起的氧化应激反应有关.     

突变SOD1 cDNA在大鼠皮层神经元定位和作用

目的:分析一个ALS家系突变铜、锌超氧化物歧化酶(Cu/Zn superoxide dismutase,SOD1)基因在原代培养的大鼠皮层神经元中的定位及作用。方法:将构建好的含有SOD1正常基因与突变基因在内的增强型绿色荧光蛋白真核表达载体(pEGFP-hSOD1和pEGFP-mSOD1)分别转染大鼠皮层神经元,转染后观察绿色荧光在细胞内的分布情况,同时设对照组,检测神经元的SOD1活性、MDA含量及细胞活性。结果:转染后荧光物质表达于胞浆内,与转染pEGFP-mSOD1质粒的神经元相比,转染pEGFP-hSOD1质粒神经元的SOD1活性、MTT值显著增高,而MDA、LDH水平显著降低。结论:突变SOD1蛋白的酶活性下降,使脂质过氧化物相对增多,导致神经元损伤,可能是该家系发病的主要原因之一。     

低氧预适应对小鼠急性脑缺血性损伤保护作用的研究

目的 探讨低氧预适应对小鼠急性脑梗死致脑缺血性损伤的保护作用.方法 将Blb/c近交系小鼠按照随机数字表法分为正常对照组(N组),不做任何处理;假手术组(C组),仅行冷光源照射,不注射玫瑰红;急性脑梗死组(CI组),光化学法诱导制作小鼠脑皮层梗死模型;低氧预适应+急性脑梗死组(HP+CI组),低氧预适应后光化学法诱导制作小鼠脑皮层梗死模型.利用行为学、免疫荧光和激光共聚焦等实验方法.分别行神经功能评定、脑梗死体积测定、细胞凋亡检测.结果 (1)脑梗死体积比较:N组、C组未发现梗死灶,CI和HP+CI组均见明显缺血梗死灶,其中HP+CI组脑梗死体积较CI组明显减小,差异有统计学意义(P<0.05);(2)神经功能评分比较:N组、C组无神经功能缺损症状,CI组和HP+CI组出现明显神经功能缺损症状,神经功能评分明显降低,其中HP+CI组神经功能评分比CI组明显增加,差异有统计学意义(P<0.05);(3)细胞凋亡比较:N组、C组小鼠大脑皮层偶见TUNEL阳性细胞,CI组和HP+CI组TUNEL阳性细胞数明显增多,其中HP+CI组TUNEL阴性细胞数较CI组减少,差异有统计学意义(P<0.05).结论 低氧预适应可能通过减小小鼠急性脑梗死体积、减少细胞凋亡两个方面发挥脑缺血性损伤的保护作用.     

腹膜透析液添加尿激酶对尿毒症并发脑梗死患者保护作用及机制

目的探讨腹膜透析液添加尿激酶对尿毒症并发脑梗死患者血清超氧化物歧化酶（SOD）、丙二醛（MDA）及血浆内皮素（ET）、一氧化氮（NO）的影响。方法将60例尿毒症并发脑梗死患者随机分为腹膜透析液添加尿激酶治疗组（30例）和常规治疗组（30例）,同时选择同期年龄在40岁以上的正常人30例为健康对照组（无肝肾脑等疾病）,两治疗组基础干预相同,尿激酶治疗组在常规治疗基础上在腹膜透析液添加尿激酶,治疗8周后观察两组患者SOD、MDA、ET、NO及临床症状的变化。用比色法测定血清MDA和SOD水平,用放射免疫法测定血浆ET的变化,NO采用硝酸还原法测定。结果①治疗前与健康对照组比较,尿激酶治疗组和常规治疗组血清SOD活性降低（P〈0．05）,NO降低（P〈0．05）,MDA含量升高（P〈0．05）,血浆内ET水平升高（P〈0．01）。②治疗8周后,常规治疗组和尿激酶治疗组可降低血浆ET水平和升高NO;与常规治疗组比较,尿激酶治疗组在降低ET和升高NO方面疗效更为显著（P〈0．01）。③常规治疗组未见SOD、MDA的变化,尿激酶治疗组能够回升SOD活性,降低MDA含量,与健康对照组及常规治疗组有明显差别（P〈0．05,P〈0．05）。结论腹膜透析液添加尿激酶可降低氧化应激反应,降低血浆ET和升高NO水平,对尿毒症并发脑梗死患者有治疗作用。     

纳洛酮对急性脑梗死的脑保护作用

目的探讨纳洛酮对急性脑梗死的脑保护作用。方法将2005-01~2010-01收治的200例急性脑梗死住院患者随机分为治疗组、对照组。对照组98例个体化常规治疗,治疗组102例在常规治疗基础上静脉应用纳洛酮,连续14 d,治疗前、治疗后第1天、3天、7天、14天进行神经功能缺损评分,评价临床疗效,观察不良反应。结果总有效率:治疗组98.0%,对照组86.7%,治疗组明显优于对照组(P<0.01);显效率治疗组95.0%,对照组68.4%,治疗组明显优于对照组(P<0.01)。治疗组无不良反应发生。结论纳洛酮治疗脑梗死,可明显改善神经功能缺损,是一种安全有效的脑细胞保护剂。     

大鼠局灶脑缺血再灌注后GM1的保护作用及其对FGFR1表达的影响

目的 探讨大鼠局灶脑缺血再灌注后神经节苷脂（ＧＭ１）的保护作用及其对碱性纤维母细胞生长因子的特异性受体（ＦＧＦＲ１）表达的影响。方法 采用检线法备大脑中动脉缺血再灌注模型。通过腹腔注射给予ＧＭ１,利用免疫组化方法检测ＦＧＦＲ１表达情况,并对病理学改变进行观察研究。结果 与对照组相比,缺血再灌组ＦＧＦＲ１表达明显增多（Ｐ〈０．０１）,而ＧＭ１用药组与缺血再灌组相比,ＦＧＦＲ１表达也明显增长（Ｐ〈０．     

PAI—1基因多态性与脑梗死及再梗死的关系

目的　探讨纤溶酶原激活物抑制物 - 1(PAI- 1)基因多态性与脑梗死及再梗死的关系。方法　对5 0例初次脑梗死 (FCI)患者、45例再次脑梗死 (RCI)患者及 6 0名健康对照者以发色底物分解法测定血浆PAI- 1活性、扩增片段长度多态性分析 PAI- 1基因启动子区 4G/ 5 G和第四内含子区 (CA) n序列多态性。结果患者血浆 PAI- 1活性 ,无论 FCI(1.13± 1.1AU/ ml)和 RCI(1.13± 0 .15 0 AU/ ml)均显著高于对照者 (0 .7±0 .2 5 AU/ m l) (均 P<0 .0 1) ;4G等位基因频率在两脑梗死组均显著高于对照者 (均 P<0 .0 1) ,且 RCI高于FCI(P<0 .0 5 ) ,并与血浆 PAI- 1活性升高及高血脂有关 ;短片段 (CA) n重复倾向于与血浆 PAI- 1活性升高相关联 ,长片段者在脑梗死特别是 RCI者中较少 ;4G等位基因的脑梗死患者中短片段 (CA) n频率较高。结论　 PAI- 1基因启动子区 4G等位基因及第四内含子区 (CA) n二核苷酸重复数少可致血浆 PAI- 1活性升高 ,从而参与脑梗死特别是 RCI的发病     

17β-雌二醇对脑缺血时Bc1-2蛋白表达的影响

目的　研究 17β 雌二醇对缺血性脑损害的神经保护作用是否与Bc1 2蛋白的表达有关。 方法　采用大鼠大脑中动脉闭塞制成局灶性脑缺血模型。在缺血 2小时 ,再灌注 2 2小时后立即断头取脑 ,应用TTC染色和免疫组化方法 ,观察脑梗死体积及Bc1 2蛋白的表达。结果　 17β 雌二醇用药组较对照组脑梗死体积显著缩小 (P <0 .0 5 ) ;Bc1 2蛋白的表达较对照组明显增强 (P <0 .0 5 )。结论　 17β 雌二醇上调大鼠脑缺血时bc1 2的表达很可能是其具有神经保护作用的机制之一。     

LIMK1mRNA及其蛋白在脆性X综合征小鼠大脑皮层中的表达

目的 观察LIMK1 mRNA及其蛋白在脆性X综合征(FXS)小鼠大脑皮层中的表达,探讨其在FXS发病机制中的作用.方法 选择雄性FMR1基因敲除型FVB近交系新生鼠和2、4、6周小鼠做为实验组(记为 KO0d、KO2W、KO4W和KO6W),雄性同龄野生小鼠(WT)作为对照组(记为WT0d、WT2W、WT4W和WT6W),每组每时间点9只.取小鼠单侧大脑皮层行LIMK1 mRNA实时荧光定量PCR分析,取另一侧大脑皮层行LIMK1蛋白Western blotting分析.结果 (1)KO6W组小鼠LIMK1 mRNA含量较同龄组WT小鼠及KO0d、KO2W和KO4W组小鼠明显升高,WT0d组小鼠LIMK1mRNA含量明显高于WT2W、WT4W和WT6W组小鼠,差异有统计学意义(P<0.05).(2)同龄KO、WT小鼠大脑皮层中LIMK1蛋白含量差异无统计学意义(P>0.05);KO0d组小鼠LIMK1蛋白含量明显低于KO2W、KO4W和KO6W组,WT0d组小鼠大脑皮层中LIMK1蛋白含量明显低于WT2W、WT4W和WT6W组,差异有统计学意义(P<0.05).结论 LIMK1蛋白的翻译过程在6周时受到明显抑制,反馈性调节转录过程使LIMK1 mRNA表达急剧升高.KO鼠LIMK1蛋白表达下降,从而影响树突棘的骨架蛋白重构,影响树突棘的功能改变,可能是FXS神经系统改变的重要机制之一.

Abstract：

Objective To observe the mRNA and protein expressions of LIMK1 in the cerebral cortex of mice with FMR1 gene knockout, and explore the roles of LIMK1 mRNA and LIMK1 protein inthe pathogenic mechanism of fragile X syndrome (FXS). Methods FVB strain male mice with FMR1 gene knockout (KO, n=36, experimental group) and their wild type (WT, n=36, control group) were equally divided into 8 counterpart subgroups (WT0d, WT2W, WT4W, WT6W, KO0d, KO2W, KO4W and KO6W),respectively, according to different ages. LIMK1 mRNA expression in the left sides of the cerebral cortex were analyzed with RT-qPCR and protein expression of LIMK1 in another side with Western blotting.Results No significant differences of LIMK1 mRNA expression in the cerebral cortex of KO0d, KO2Wand KO4W subgroups were noted as compared with that of respective age-matched WT mice (P>0.05);but that of KO6W subgroup was significantly increased as compared with that of WT6W subgroup, and KO0d,KO2W and KO4W subgroups (P<0.05);the level of LIMK1 mRNA in WT0d subgroup was obviously higher than that of WT2W, WT4W and WT6W subgroups (P<0.05). No statistic differences of LIMK1 protein between the same-age KO and WT mice were noted (P>0.05);significantly lower level of LIMK1 protein in KO0d subgroup was found as compared with that of KO2W, KO4W and KO6W subgroups (P<0.05);that of WT0d subgroup was lower than that of WT2W, WT4W and WT6W subgroups (P<0.05). Conclusion The translation process of LIMK1 protein is significantly inhibited at 6 weeks and LIMK1 mRNA expression increased sharply based on the feedback adjustment of LIMK1 protein expression declining;once this translation process is inhibited or interrupted, it will affect the dendritic spines skeleton protein reconstruct and lead to the dendritic spines function deficient;the inhabitation of translation process might probably play an important role in the process of dendritic spines maturation and should be an important pathogenic mechanism of FXS.     

Protective effect of oxysophoridine on cerebral ischemia/reperfusion injury in mice

Oxysophoridine, a new alkaloid extracted from Sophora alopecuroides L., has been shown to have a protective effect against ischemic brain damage. In this study, a focal cerebral ischemia/reperfusion injury model was established using middle cerebral artery occlusion in mice. Both 62.5, 125, and 250 mg/kg oxysophoridine, via intraperitoneal injection, and 6 mg/kg nimodipine, via intragastric administration, were administered daily for 7 days before modeling. After 24 hours of reperfusion, mice were tested for neurological deficit, cerebral infarct size was assessed and brain tissue was collected. Results showed that oxysophoridine at 125, 250 mg/kg and 6 mg/kg nimodipine could reduce neurological deficit scores, cerebral infarct size and brain water content in mice. These results provided evidence that oxysophoridine plays a protective role in cerebral ischemia/reperfusion injury. In addition, oxysophoridine at 62.5, 125, and 250 mg/kg and 6 mg/kg nimodipine increased adenosine-triphosphate content, and decreased malondialdehyde and nitric oxide content. These compounds enhanced the activities of glutathione-peroxidase, superoxide dismutase, catalase, and lactate dehydrogenase, and decreased the activity of nitric oxide synthase. Protein and mRNA expression levels of N-methyl-D-aspartate receptor subunit NR1 were markedly inhibited in the presence of 250 mg/kg oxysophoridine and 6 mg/kg nimodipine. Our experimental findings indicated that oxysophoridine has a neuroprotective effect against cerebral ischemia/reperfusion injury in mice, and that the effect may be due to its ability to inhibit oxidative stress and expression of the N-methyl-D-aspartate receptor subunit NR1.     

丹参素预处理对脑梗死大鼠血浆纤溶激活系统的影响

目的 观察丹参素预处理对脑梗死大鼠血浆纤溶激活系统的影响.方法 40只雄性大鼠,随机分为假手术组、脑梗死对照组、低剂量[5 mg/(kg·d)]丹参素组和高剂量[10 mg/(kg·d)]丹参素组,每组10只.应用自体血栓注入,制成大鼠脑梗死模型;用ELISA方法 检测血浆中组织型纤溶酶原激活物(t-PA)及其抑制物(PAI-I)含量;RT-PCR测定大鼠脑组织中t-PA mRNA及其PAI-I mRNA表达的变化.结果 与脑梗死对照组[(2.65±0.43)ng/ml]比较,低剂量及高剂最丹参素组t-PA含量[(3.29±0.13)ng/ml、(6.37±0.19)ng/ml]明显升高(均P＜0.01),两丹参素组之间差异亦有统计学意义(P＜0.0I);与脑梗死对照组比较,低剂量丹参素组PAI-I含量[(2.65±0.40)ng/ml]降低(P＜0.05),高剂量丹参素组PAI-I含量[(1.98±0.32)ng/ml]明显降低(P＜0.01),两丹参素组之间差异亦有统计学意义(P＜0.01);与脑梗死对照组比较,低剂量及高剂量丹参素组t-PA mRNA表达[灰度值分别为(0.37±0.14)、(0.46±0.12)]均明显升高(均P＜0.01),两丹参素组之间差异亦有统计学意义(P＜0.01);与脑梗死对照组比较,低剂量及高剂量丹参素组PAI-I mRNA表达[灰度值分别为(0.54±0.12)、(0.36±0.07)]均明显降低(均P＜0.01),两丹参素组之间差异亦有统计学意义(P＜0.01).结论 丹参素可以改善脑梗死大鼠纤溶功能,并且有剂量依赖性.     

H2S对急性脑梗死大鼠神经功能的保护作用及机制

目的 探讨H2S对急性脑梗死大鼠神经功能的保护作用以及相关机制.方法 将48只健康雄性SD大鼠随机分为假手术组、模型组、生理盐水组和H2S干预组.采用改良Longa法制备大鼠急性脑梗死模型,建模前25min,H2S干预组给予腹腔注射NaHS 28μmol/kg,生理盐水组给予等量生理盐水,分别于苏醒后和术后72h,评估各组大鼠的神经功能缺损.采用TTC染色法检测各组大鼠脑梗死面积,TUNEL法检测各组大鼠脑组织中细胞凋亡情况,Western blot法检测各组大鼠脑组织中PI3K、p-PI3K、Akt、p-Akt、Bcl-2、Bax蛋白的表达.结果 与模型组和生理盐水组相比,H2S干预组大鼠神经功能缺损程度评分、脑梗死面积和细胞凋亡指数均降低,差异均有统计学意义(P<0.05).与模型组和生理盐水组相比,H2S干预组大鼠脑组织中PI3K、Akt、Bcl-2蛋白相对表达量均升高,而p-PI3K、p-Akt、Bax蛋白相对表达量均降低,差异均有统计学意义(P<0.05).结论 H2S可能通过抑制PI3K/Akt信号通路而改变Bcl-2/Bax比例,减小急性脑梗死大鼠脑梗死面积及细胞凋亡指数,保护神经功能.     

Protective effect of the ketogenic diet in Scn1a mutant mice

Purpose: We evaluated the ability of the ketogenic diet (KD) to improve thresholds to flurothyl‐induced seizures in two mouse lines with Scn1a mutations: one that models Dravet syndrome (DS) and another that models genetic (generalized) epilepsy with febrile seizures plus (GEFS+). Methods: At postnatal day 21, mouse models of DS and GEFS+ were fasted for 12–14 h and then placed on either a 6:1 (fats to proteins and carbohydrates) KD or a standard diet (SD) for 2 weeks. At the end of the 2‐week period, we measured thresholds to seizures induced by the chemiconvulsant flurothyl. Body weight, β‐hydroxybutyrate (BHB) levels, and glucose levels were also recorded every 2 days over a 2‐week period in separate cohorts of mutant and wild‐type mice that were either on the KD or the SD. Key Findings: Mice on the KD gained less weight and exhibited significantly higher BHB levels compared to mice on the SD. It is notable that thresholds to flurothyl‐induced seizures were restored to more normal levels in both mouse lines after 2 weeks on the KD. Significance: These results indicate that the KD may be an effective treatment for refractory patients with SCN1A mutations. The availability of mouse models of DS and GEFS+ also provides an opportunity to better understand the mechanism of action of the KD, which may facilitate the development of improved treatments.