肢体缺血后处理对糖尿病大鼠脑缺血再灌注损伤线粒体结构和功能的影响

目的探讨肢体缺血后处理(I-PostC)诱导的远隔器官I-PostC(RPostC)对糖尿病大鼠局灶性脑缺血再灌注(I/R)损伤线粒体结构和功能的影响。方法链脲佐菌素空腹腹腔注射制作糖尿病大鼠模型,线栓法闭塞大脑中动脉(MCAO)制作局灶性I/R大鼠模型。造模成功的40只雄性SD大鼠分为4组:空白对照组、假手术组、I/R组,RPostC组,每组10只。I/R6h后取脑组织行HE染色观察,测定线粒体丙二醛(MDA)含量、超氧化物歧化酶(SOD)、Na+/K+-ATP酶、Ca2+-ATP酶和谷胱甘肽过氧化物酶(GSH-Px)活性,观察线粒体超微结构的改变。结果I/R组线粒体MDA含量[(4.99±1.25)nmol/mgprot]较空白对照组和假手术组明显升高(均P<0.01),SOD[(72.52±13.07)U/mg]、Na+/K+-ATP酶[(3.17±0.34)μmolPi/(mg.h)]、Ca2+-ATP酶[(1.56±0.23)μmolPi/(mg.h)]和GSH-Px活性[(22.66±5.29)U/mg)]明显降低(均P<0.01);与I/R组比较,RPostC组MDA含量[(3.58±0.91)...     

葛根素预处理对大鼠局灶性脑缺血-再灌注线粒体功能的影响

目的观察葛根素对大鼠局灶性脑缺血-再灌注损伤后线粒体功能的影响。方法健康成年雄性SD大鼠40只,随机分成5组(n=8):假手术组、脑缺血-再灌注组、葛根素预处理24h 100mg组、葛根素预处理24h 200mg组和葛根素预处理24h 400mg组,其中葛根素预处理24h组于脑缺血前24h给予相应剂量葛根素腹腔注射行单次预处理,采用大脑中动脉线栓法建立局灶性脑缺血-再灌注模型,缺血90min,再灌注24h后,测定缺血侧海马线粒体内游离MDA、SOD、Na+-K+-ATP酶及Ca2+-ATP酶活性。结果与脑缺血-再灌注组相比,葛根素预处理-24 h100mg组、葛根素预处理-24h 200mg组及葛根素预处理-24h 400mg组大鼠脑线粒体内MDA含量明显下降,SOD、Na+-K+-ATPase、Ca2+-ATPase活性明显升高(P<0.05),而在葛根素预处理3个剂量组间比较差异均无统计学意义(P﹥0.05)。结论葛根素预处理对脑缺血-再灌注后线粒体损伤具有非剂量依赖性保护作用。     

桃仁红花煎剂对大鼠缺血再灌注后脑组织的保护作用

目的研究桃仁红花煎剂对大鼠局灶性脑缺血再灌注后脑组织的影响。方法 45只雄性SD大鼠随机平均为给药组、模型组和对照组。采用线栓法阻塞大鼠右侧大脑中动脉,使其缺血2h后再灌注24h建立局灶性脑缺血再灌注模型。术前2h和术后3、12h分3次灌胃给予桃仁红花煎剂,总剂量是40g/kg。通过神经行为评分评定大鼠神经功能变化,按干湿重法测定脑含水量,用氯化三苯基四氮唑法测定脑梗死范围,分光光度法测定缺血区脑组织中Na+-K+-ATP酶和Ca2+-ATP酶的活性。结果在缺血再灌注3h和24h后,给药组神经行为评分明显高于模型组(P<0.05)。缺血再灌注24h,给药组脑含水量和脑梗死体积明显少于模型组(P<0.05);给药组缺血脑皮层中Na+-K+-ATP酶和Ca2+-ATP酶活性明显高于模型组(P<0.05)。结论桃仁红花煎剂对大鼠缺血再灌注后脑组织有保护作用,其机制可能与其增强Na+-K+-ATP酶和Ca2+-ATP酶的活性、减轻脑水肿有关。     

异丙酚对大鼠脑缺血再灌注损伤线粒体能量代谢的影响

目的 探讨静脉麻醉药异丙酚对脑缺血再灌注损伤模型大鼠的神经保护作用.方法 SD大鼠48只按随机数字表法分为假手术组、模型组、50 mg/kg异丙酚和100 mg/kg异丙酚组,每组12只.后3组大鼠均制备脑缺血(2 h)再灌注(24 h)模型,恢复灌注后分别经腹腔内注射等体积生理盐水和50、100 mg/kg异丙酚.再灌注24 h后对大鼠进行神经功能缺损评分,应用TTC染色观察脑梗死的范围,化学比色法检测脑组织线粒体琥珀酸脱氢酶(SDH)、Na+-K+-ATP酶、Ca2+-Mg2+-ATP酶活性的改变.结果 与模型组比较,50、100mg/kg异丙酚组大鼠神经功能缺损评分较低,脑组织线粒体SDH、Na+-K+-ATP酶、Ca2+Mg2+-ATP酶活性增高,差异有统计学意义(P＜0.05);与50mg/kg异丙酚组[(45.9±25.1)U/mg pro,(5.24±0.85)分]比较,100mg/kg异丙酚组SDH活性[(96.1±20.8)U/mgpro]较高,神经功能缺损评分(4.40±0.79)较低,差异有统计学意义(P＜0.05).结论异丙酚对脑缺血再灌注损伤大鼠具有神经保护作用,促进线粒体SDH、Na+-K+-ATP酶、Ca2+Mg2+-ATP酶活性的恢复、保护线粒体功能是其机制之一.     

有氧运动对大鼠骨骼肌线粒体Na+，K+-ATP酶和Ca2+-ATP酶及线粒体肿胀的影响

背景：研究表明有氧运动可提高线粒体功能,但在不同时期的作用特点还不明确。 目的：观察不同周期有氧运动对大鼠骨骼肌线粒体Na+,K+-ATP酶和Ca2+-ATP酶以及线粒体肿胀的影响。 方法：将SD大鼠随机分为正常对照组,有氧运动2,4和6周组。正常对照组不进行有氧运动,其余3组则参照BedfordTG标准,采用跑台运动方式,建立有氧运动模型进行相应的运动周期锻炼。测定各组大鼠骨骼肌线粒体Na+,K+-ATP酶和Ca2+-ATP酶的活性以及线粒体肿胀程度。 结果与结论：有氧运动2周组各指标与对照组比较无差异。有氧运动4和6周组线粒体Na+,K+-ATP酶、Ca2+-ATP酶活性均增高(P < 0.05),线粒体肿胀程度降低(P < 0.05)。实验结果表明,有氧运动可保护线粒体Na+,K+-ATP酶、Ca2+-ATP酶的活性,提高线粒体功能,但需要一定的时间积累。     

原花青素对大鼠脑缺血再灌注损伤代谢障碍的影响

目的 探讨原花青素(PC)对大鼠脑缺血再灌注损伤代谢障碍的影响.方法 采用Zea-Longa线栓法制备大鼠局灶性脑缺血再灌注模型.术前0.5 h和脑缺血2 h时给予大鼠PC溶液50、100、200 mg/kg,假手术组和模型组分别给予等量的生理盐水.观察不同剂量PC溶液对大鼠脑缺血再灌注后神经功能状态、脑组织含水量及乳酸含量、能量代谢酶(Na+-K+-ATP酶和Ca2+-ATP酶)活性的影响.结果 PC中、高剂量组神经功能行为学评分和脑组织含水量显著低于模型组(P＜0.05).PC低、中、高剂量组脑组织乳酸含量显著低于模型组(P＜0.05).PC中、高剂量组Na+-K+-ATPase活性较模型组升高(P＜0.05).PC高剂量组Ca2+-ATPase活性较模型组有所增高(P＜0.05).结论 PC可通过减轻脑水肿、改善脑组织的代谢障碍而发挥脑保护作用.     

骨骼肌肌浆网Na+，K+-ATP酶和Ca2+-ATP酶活性在不同训练条件下的变化

背景：Na+,K+-ATP酶和Ca2+-ATP酶在物质运送、能量转换以及信息传递方面具有重要作用。肌浆网在肌肉兴奋-收缩耦联过程中起关键作用,与运动性骨骼肌疲劳的发生密切关。 目的：通过建立SD大鼠有氧和无氧训练模型,观察不同训练负荷条件对大鼠骨骼肌肌浆网Na+,K+-ATP酶和Ca2+-ATP酶活性的影响。 方法：参照Bedford TG标准,建立有氧和无氧运动大鼠跑台训练模型,有氧运动组采用递增负荷训练,无氧运动组采用高速间歇训练,正常对照组大鼠正常笼内生活,不运动。各组动物训练结束后用超速离心法提取大鼠骨骼肌肌浆网,紫外分光光度计检测大鼠骨骼肌肌浆网Na+,K+-ATP酶和Ca2+-ATP酶的活性。 结果与结论：训练4周后,两个运动组大鼠骨骼肌肌浆网Na+,K+-ATP酶和Ca2+-ATP酶的活性逐渐升高(P < 0.05);训练6周,仅有氧运动组升高(P < 0.05),无氧运动组则活性降低(P < 0.05)。结果提示有氧训练更有利于保护大鼠骨骼肌肌浆网Na+,K+-ATP酶和Ca2+-ATP酶的活性,但需要一定的时间累积。     

肠淋巴再灌注加重肠系膜上动脉闭塞性休克大鼠的脑损伤*

目的：观察肠淋巴再灌注（MLR）对肠系膜上动脉闭塞性（SMAO）休克大鼠脑组织形态学的影响；从氧自由基、一氧化氮、中性粒细胞、膜泵、能量代谢等方面揭示其机制。方法：24只Wistar雄性大鼠均分为4组：Sham组，仅麻醉与手术；MLR组，夹闭肠系膜淋巴管（ML）1h，再灌注2h；SMAO组，夹闭肠系膜上动脉（SMA）1h，再灌注2h；MLR+SMAO：夹闭ML和SMA1h，再灌注2h。于再灌注2h后，选择固定位置留取脑组织，制备病理切片，观察形态学；同时制备脑组织匀浆，用于检测LA、MDA、SOD、NO、NOS、MPO、ATPase及ATP。结果：Sham与MLR组大鼠脑组织结构基本正常；SMAO组大鼠可见神经元有坏死、变性，偶见肿胀；MLR+SMAO组神经元损伤情况较SMAO组重。MLR与Sham组脑组织匀浆各项指标均无统计学差异；SMAO与MLR+SMAO组脑匀浆MDA、NO、LA含量、NOS与MPO活性均显著高于MLR与Sham组，且MLR+SMAO组脑匀浆MDA、NO含量、NOS与MPO活性均显著高于SMAO组；SMAO组脑匀浆SOD、Na+- K+-ATPase活性显著低于Sham与MLR组、Mg2+-ATPase活性、ATP含量显著低于MLR组；MLR+SMAO组脑匀浆的SOD、Na+- K+-ATPase、Ca2+-ATPase、Mg2+-ATPase及Ca2+-Mg2+- ATPase活性均显著低于Sham与MLR组，且Ca2+-ATPase、Mg2+-ATPase及Ca2+-Mg2+-ATPase活性、ATP含量均显著低于SMAO组。结论：MLR加重SMAO休克大鼠的脑损伤，其机制可能与MLR加重或增加脑组织氧自由基损伤、NO合成与释放、中性粒细胞扣押、能量代谢障碍及降低脑组织细胞膜泵活性等因素有关。     

大鼠脑缺血再灌注后线粒体ATP酶活性、形态学变化及丹参多酚酸的保护作用

目的观察注射用丹参多酚酸对大鼠脑缺血再灌注损伤后线粒体ATP酶活性、及形态学的影响。方法将54只雄性SD大鼠随机分成3组(n=18):假手术组(Sham组)、缺血再灌注组(IR组)、丹参多酚酸组(Salvianolate组)。HE染色法观察大脑神经元的组织病理学变化;TTC染色检测脑梗死体积;分光光度计发法线粒体丙二醛含量和线粒体Na~+/K~+ATP酶、Ca~(2+)ATP酶、Mg~(2+)ATP酶活性。结果假手术组大鼠脑组织红染,未见梗死灶;丹参多酚酸组神经细胞变性坏死程度、脑梗死面积较缺血再灌注组明显减轻;与缺血再灌注组相比,丹参多酚酸组线粒体丙二醛含量下降(P0.05)和线粒体Na~+/K~+ATP酶、Ca~(2+)ATP酶、Mg~(2+)ATP酶活性升高(P0.05)。结论注射用丹参多酚酸对缺血再灌注损伤有保护作用,其机制与提高线粒体ATP酶活性有关。     

注射用丹参多酚酸对大鼠脑缺血再灌注线粒体ATP酶活性的影响

目的探讨注射用丹参多酚酸对大鼠脑缺血再灌注线粒体ATP酶活性的影响。方法将54只雄性SD大鼠随机分成3组(n=18):假手术组(Sham组)、缺血再灌注组(IR组)、丹参多酚酸组(Salvianolate组)。HE染色法观察大脑神经元的组织病理学变化;TTC染色检测脑梗死体积;分光光度计发法线粒体丙二醛含量和线粒体Na~+/K~+ATP酶、Ca~(2+)ATP酶、Mg~(2+)ATP酶活性。结果假手术组大鼠脑组织红染,未见梗死灶;丹参多酚酸组神经细胞变性坏死程度、脑梗死面积较缺血再灌注组明显减轻;与缺血再灌注组相比,丹参多酚酸组线粒体丙二醛含量下降(P0.05)和线粒体Na~+/K~+ATP酶、Ca~(2+)ATP酶、Mg~(2+)ATP酶活性升高(P0.05)。结论注射用丹参多酚酸对缺血再灌注损伤有保护作用,其机制与提高线粒体ATP酶活性有关。     

Cromakalin pretreatment affects mitochondrial structure and function in a rat model of ischemia/reperfusion injury

BACKGROUND: Mitochondrial structural changes and energy dysmetabolism frequently occur subsequent to cerebral ischemia. Adenosine triphosphate (ATP)-sensitive potassium channel openers exhibit protective effects on cerebral ischemia/reperfusion injury. OBJECTIVE: To validate the effects of cromakalin on mitochondrial structure and function in ischemic penumbra brain tissue in a rat model of middle cerebral artery occlusion (MCAO). DESIGN, TIME AND SETTING: The present single-factor analysis of variance, randomized, controlled, animal experiment was performed at the Institute of Brain Science, Affiliated Hospital of Qingdao University Medical College between October 2007 and March 2008. MATERIALS: Forty male, Wistar rats were randomly divided into four groups, with 10 rats per group: sham-operated, MCAO, MCAO ATP-sensitive potassium channel opener (cromakalin), and MCAO eromakalin ATP-sensitive potassium channel blocking agent (glibenclamide). METHODS: Focal cerebral ischemia/reperfusion injury was induced by MCAO in all groups except the sham-operated group. The MCAO cromakalin group was administered 10 mg/kg cromakalin (i.p.) prior to MCAO induction. The MCAO cromakalin glibenclamide group received an injection of 10 mg/kg cromakalin (i.v.), and subsequently an injection of 10 mg/kg cromakalin (i.p.) prior to MCAO induction. MAIN OUTCOME MEASURES: At 24 hours after cerebral ischemia/reperfusion injury, cellular apoptosis was detected by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) nick-end labeling technique. Cytochrome C expression was measured by immunohistochemistry. In addition, mitochondrial swelling, membrane fluidity, membrane phospholipid and malonaldehyde (MDA) contents, as well as Na -K -ATPase, Ca2 -ATPase, and superoxide dismutase (SOD) activities were determined. RESULTS: Compared with the sham-operated group, the three ischemia groups exhibited significantly elevated mitochondrial MDA content, reduced membrane phospholipid and ATP contents, down-regulated membrane fluidity, and reduced Na -K -ATPase, Ca2 -ATPase, and SOD activities (P < 0.05-0.01 ). In the MCAO eromakalin group, the number of apoptotic cells decreased, and cytochrome C expression, as well as MDA content, were reduced. However, ATP content and Na -K -ATPase, Ca2 -ATPase, and SOD activities significantly increased compared with the MCAO group (P < 0.05-0.01 ). Glibenclamide noticeably antagonized cromakalin protection of mitochondria.CONCLUSION: Pretreatment with the ATP-sensitive potassium channel opener cromakalin increased mitochondrial Na -K -ATPase, Ca2 -ATPase, and SOD activities, decreased neuronal apoptosis, and inhibited cytochrome C expression following MCAO.     

促红细胞生成素在脑缺血再灌注损伤中对线粒体的保护作用

目的 探讨促红细胞生成素(EPO)在脑缺血再灌注损伤中对线粒体功能的保护作用. 方法 30只SD大鼠按随机数字表法分为正常对照组、缺血再灌注组和EPO治疗组3组,每组各10只.EPO治疗组和缺血再灌注组用线栓法复制脑缺血再灌注模型.EPO治疗组在缺血再灌注后1、24、48、60 h腹腔注射EPO,剂量为3000 U/kg(用生理盐水以1:1比例稀释),缺血再灌注组腹腔注射同等剂量的生理盐水.正常对照组仅分离颈部动脉,动脉不做栓塞处理.缺血后72h观察各组大鼠脑组织神经细胞线粒体跨膜电位、线粒体丙二醛、超氧化物歧化酶、Na十_K+.ATP酶活性、一氧化氮含量和免疫组化检测海马区Caspase-3阳性细胞数的变化. 结果 EPO治疗组的神经细胞线粒体跨膜电位(77.48±5.93)、超氧化物歧化酶[(96.91±8.66)p,kat/g]、Na+_K+-ATP酶活性[(10.48±2.77)μkat/g]明显高于缺血再灌注组[44.47±17.35、(84.46±8.54)μkat/g、(7.37±2.87)μkat/g],线粒体丙二醛[(37.99±5.38)μmol/g]、一氧化氮含量[(10.18±2.02)μmol/g]、Caspase-3阳性细胞数(66.31±8.09)明显低于缺血再灌注组[(44.83±6.48)μmol/g、(12.12±2.14)μmol/g、74.90±7.42]. 结论 EPO对缺血再灌注损伤脑组织产生保护作用的重要机制之一是保护神经细胞线粒体的功能,其核心是抑制线粒体跨膜电位的下降.     

羧乙基锗倍半氧化物对大鼠脑缺血再灌注损伤的保护作用

采用结扎双侧颈总动脉后再通的方法复制大鼠脑缺血再灌注损伤模型，通过测定再灌注后大鼠海马组织中脂质过氧化产物丙二醛（ＭＤＡ），超氧化物歧化酶（ＳＯＤ）与谷胱甘肽过氧化物酶（ＧＳＨ—Ｐｘ）及ＡＴＰａｓｅ的活性，观察了有机锗─羧乙基锗倍半氧化物（ＣＧＳ）对大鼠脑缺血再灌注后大鼠海马组织中ＭＤＡ水平，明显保护ＳＯＤ、ＧＳＨ─Ｐｘ、Ｎａ＋Ｋ＋─ＡＴＰａｓｅ及Ｃａ２＋─ＡＴ─Ｐａｓｅ活性。表明ＣＧＳ对大鼠脑缺血再灌注损伤具有保护作用。     

maFGF对慢性衰老大鼠脑组织中ATP酶活力的影响

目的观察改构型酸性成纤维细胞生长因子(maFGF)对D-半乳糖致衰老大鼠脑组织中Na+-K+-ATP酶活力及Ca2+-Mg2+-ATP酶活力的影响,探讨maFGF抗衰老的机制。方法选择成年Wistar大鼠40只,采用皮下注射D-半乳糖建立衰老模型,衰老模型成功后随机分为衰老对照组、NS对照组和maFGF治疗组。另10只不注射D-半乳糖作为正常对照组。maFGF治疗组按12μg/kg剂量肌内注射,1次/d,共14 d,NS对照组肌肉注射与maFGF治疗组相同容量的生理盐水,1次/d;衰老对照组不作任何干预。各组大鼠到相对应的时间点取出各组大鼠脑组织,测定脑组织匀浆中Na+-K+-ATP酶活力及Ca2+-Mg2+-ATP酶活力。结果与正常对照组相比,衰老对照组大鼠脑组织中Na+-K+-ATP酶活力及Ca2+-Mg2+-ATP酶活力均明显著降低(P<0.01);maFGF治疗组脑组织中Na+-K+-ATP酶活力和Ca2+-Mg2+-ATP酶活力均明显高于衰老对照组、NS对照组(P<0.05)。结论 maFGF能升高衰老大鼠脑组织中Na+-K+-ATP酶活力及Ca2+-Mg2+-ATP酶活力,对衰老大鼠脑损伤有一定保护作用。     

Effects of the hyperbaric oxygen treatment on the Na+,K+ -ATPase and superoxide dismutase activities in the optic nerves of global cerebral ischemia-exposed rats

The effects of hyperbaric oxygen (HBO) treatment on the Na+,K+ -ATPase and superoxide dismutase (SOD) activities were examined in the optic nerves of the rats exposed to global cerebral ischemia. Animals were exposed to global cerebral ischemia of 20-min duration and were either sacrificed or exposed to the first HBO treatment immediately, 0.5, 1, 2, 6, 24, 48, 72 or 168 h after ischemic procedure (for Na+,K+ -ATPase activities measurement) or 2, 24, 48 or 168 h after ischemia (for SOD activities measurement). HBO procedure was repeated for 7 consecutive days. It was found that global cerebral ischemia induced a statistically significant decrease in the Na+,K+ -ATPase activity of the optic nerves, starting from 0.5 to 168 h of reperfusion. Maximal enzymatic inhibition was registered 24 h after the ischemic damage. The decline in the Na+,K+ -ATPase activity was prevented in the animals exposed to HBO treatment within the first 6 h of reperfusion. The results of the presented experiments demonstrated also a statistically significant increase in the SOD activity after 24, 48 and 168 h of reperfusion in the optic nerves of non-HBO-treated ischemic animals as well as in the ischemic animals treated with HBO. Our results indicate that global cerebral ischemia induced a significant alterations in the Na+,K+ -ATPase and SOD activities in the optic nerves during different periods of reperfusion. HBO treatment, started within the first 6 h of reperfusion, prevented ischemia-induced changes in the Na+,K+ -ATPase activity, while the level of the SOD activity in the ischemic animals was not changed after HBO administration.     

硫酸镁对大鼠脑缺血再灌流损伤保护作用的研究

目的:探讨ＭｇＳＯ４对大鼠脑缺血再灌流损伤的保护作用。方法:线栓法将ＳＤ大鼠制成脑缺血再灌流模型,分组测定脑组织Ｎａ＋－Ｋ＋－ＡＴＰａｓｅ活性、ＭＤＡ、ＮＯ含量。结果:治疗１组(３００ｍｇ·ｋｇ－１)和治疗２组(６００ｍｇ·ｋｇ－１)与对照组比较,Ｎａ＋－Ｋ＋－ＡＴＰａｓｅ活性增加,ＭＤＡ、ＮＯ含量降低,且治疗２组比治疗１组作用更明显。结论:ＭｇＳＯ４对脑缺血再灌流损伤有保护作用,６００ｍｇ·ｋｇ－１较３００ｍｇ·ｋｇ－１更显著。     

Influence of lactate accumulation on Na,K-ATPase activity of ischemic and post-ischemic brain

In this study the cerebral Na+, K+-ATPase activity as well as selected parameters of oxidative metabolism and electrophysiological function were assessed in normoglycemic and hyperglycemic rats which were exposed to ischemia produced by electrocautery of the vertebral arteries and reversible occlusion of the carotid arteries. In hyperglycemic animals 0.5 h of ischemia was associated with massive accumulation of lactate (34 mumol X g-1) and enhanced Na+, K+-ATPase activity (116% control), whereas normoglycemic animals showed more moderate lactate accumulation (17 mumol X g-1) and normal Na+, K+-ATPase activity (102% control). In normoglycemic animals release of the carotid clamps and recirculation for 0.5-1.5 h was associated with a normalization of the lactate levels and a decrease in Na+, K+-ATPase activity (68-72% control). Restituted hyperglycemic animals showed metabolic changes which seemed related to the blood pressure, with hypotensive hyperglycemic animals showing continuing massive lactacidosis (30-35 mumol X g-1) and enhanced Na+, K+-ATPase activity (108-110% control), whereas normotensive hyperglycemic animals showed progressive decreases in lactate level (14-20 mumol X g-1) and normal or mildly suppressed Na+, K+-ATPase activity (88-97% control). These patterns of change suggest that the reperfusion of the post-ischemic hyperglycemic-hyperlactacidotic brain was inadequate or non-homogeneous.