EGb761对大鼠局灶性脑缺血再灌注损伤后细胞凋亡的影响

本实验用线栓法建立大鼠局灶性脑缺血再灌注损伤模型 ,观察银杏叶提取物的标准制剂 EGb76 1(含 2 4 %黄酮甙和 6 %萜烯 )对细胞凋亡以及脑缺血后脑组织病理变化的影响。1　材料与方法1.1　实验动物分组　健康 Wistar大鼠 2 0只 ,体重 2 5 0～ 30 0 g,随机分为假手术组 (A组 ) ,脑缺血再灌注组 (B组 ) ,生理盐水组治疗组 (C组 ) ,EGb76 1治疗组 (D组 ) ,每组 5只。1.2　实验方法　线栓法制作大鼠脑缺血再灌注损伤模型 ,假手术组仅分离血管 ,不插尼龙线。 D组于实验前 3d开始灌胃给 EGb76 1(Ginaton) ,15 0 mg/ kg,每日 2次 ,术前 1h灌…     

西比灵对鼠脑缺血再灌注损伤模型的血清及脑组织中SOD、MDA、N0、NOS含量及病理学的影响

目的 研究西比灵对鼠脑缺血再灌注损伤模型的血清及脑组织中超氧化物歧化酶(SOD)、丙二醛(MDA)、一氧化氮(NO)、一氧化氮合酶(NOS)的含量及病理学改变的影响。方法 选用21只沙土鼠并随机分为3组,脑缺血再灌注组(再灌注组)、西比灵预防组(预防组)及对照组,每组7只,分别测定血清、脑组织中SOD、MDA、NO、NOS的含量,并用电镜检测其病理学的变化。结果 与对照组比较,再灌注组显示严重的病理损害,其血清、脑组织中MDA、N0、NOS含量明显上升、SOD含量明显下降,两组比较有显著性差异(均P<0.01);与再灌注组比,预防组病理改变较轻,其血清、脑组织中MDA、NO、NOS明显下降,SOD明显上升,两组比较均有显著性差异(分别P<0.01和P<0.05)。结论 西比灵可减少自由基,诱导SOD生成,具有保护脑细胞的作用。     

山楂总黄酮对实验性脑缺血再灌注损伤大鼠的保护作用

目的 研究山楂总黄酮（HFs）对实验性脑缺血再灌注损伤大鼠的保护作用.方法 SD雄性大鼠90只随机分为5组,各组按规定预防给药15 d后,按照Longa法制作大脑中动脉闭塞模型,假手术组除不插入栓线外其余相同,缺血2 h后再灌注.分别于再灌注3 h、24 h进行神经行为评分;再灌注24 h后断头取梗死侧大脑皮质匀浆,测定丙二醛（MDA）、一氧化氮（NO）含量及超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、一氧化氮合成酶（NOS）、诱导型一氧化氮合成酶（iNO）活性.结果 山楂总黄酮各剂量组再灌注3 h、24 h神经行为评分明显低于缺血再灌注组;与I/R组比较,再灌注24 h后,山楂总黄酮各剂量组缺血梗死侧大脑皮质丙二醛（MDA）、一氧化氮（NO）含量明显降低,超氧化物歧化酶（SOD）、过氧化氢酶（CAT）活性升高,一氧化氮合成酶（NOS）、诱导型一氧化氮合成酶（iNO）活性也明显低于I/R组,各组间差异有统计学意义.结论 山楂总黄酮对实验性脑缺血再灌注损伤大鼠有一定的保护作用,其作用机制可能与提高脑组织中SOD和CAT活性、抑制脂质过氧化及炎症反应有关.     

人参总皂甙对大鼠脑缺血再灌注后MDA、SOD及细胞凋亡的影响

目的:观察人参总皂甙对大鼠脑缺血再灌注的保护作用,探讨其作用机制。方法:将40只大鼠随机分为4 组:假手术组、缺血再灌注组、治疗组1、治疗组2,采用线栓法制备大鼠脑缺血再灌注模型,72h断头取脑,Nissel染色光镜下观察海马CA1区病理形态变化,TUNEL法检测细胞凋亡,同时检测脑组织中丙二醛(MDA)、超氧化物歧化酶(SOD)的含量。结果:与缺血再灌注组相比,人参总皂甙治疗组光镜下病理损伤轻,脑组织中MDA含量降低、SOD含量升高,细胞凋亡数降低。结论:人参总皂甙对大鼠脑缺血再灌注损伤具有保护作用,其机制可能与抑制自由基损伤有关。     

bFGF对大鼠局灶性缺血再灌注脑组织的保护作用

目的探讨bFGF对大鼠脑缺血再灌注损伤后神经细胞凋亡和脑组织中SOD、MDA含量变化的影响。方法应用线栓法制作大鼠局灶性脑缺血再灌注模型,大脑中动脉阻塞1h再灌注损伤24h,检测假手术组、缺血再灌注组和bFGF组的凋亡细胞数和脑组织中SOD、MDA的含量。结果假手术组偶见凋亡细胞,缺血再灌注组和bFGF组凋亡细胞数分别是26.35±5.67和18.65±5.91,与缺血再灌注组相比,bFGF组缺血区皮质凋亡神经元明显减少(P<0.05)。SOD,MDA测定结果显示三组间比较,具有显著性差异(P<0.01和P<0.05)。结论抗氧化作用可能是bFGF减少大鼠脑缺血再灌注损伤后细胞凋亡的分子机制之一。     

依达拉奉对脑缺血再灌注大鼠脑组织及血清超氧化物歧化酶、一氧化氮、丙二醛水平的影响

目的探讨依达拉奉对局灶性脑缺血再灌注损伤的影响。方法将SD大鼠分为假手术组,脑缺血再灌注组和依达拉奉干预组(干预组),采用线栓法制备大脑中动脉缺血再灌注模型;缺血1h后,设再灌注2h、6h、12h、24h组,采用化学比色法检测各组脑组织及血清超氧化物歧化酶(SOD)、一氧化氮(NO)、丙二醛(MDA)浓度。结果缺血再灌注组脑组织SOD下降,血清SOD先升后降;脑组织NO浓度先降后升,血清NO浓度持续升高;脑组织及血清MDA浓度均先升后降;与缺血再灌注组比,干预组SOD下降幅度小(均P<0·01),NO、MDA浓度明显降低;干预组6h、12h脑组织含水量明显低于缺血再灌注组(均P<0·01)。结论依达拉奉可降低羟自由基水平,对脑缺血再灌注损伤有保护作用。     

西比灵对鼠脑缺血再灌注损伤模型的血清及脑组织中SOD、MDA、NO、NOS含量及病理学的影响

目的研究西比灵对鼠脑缺血再灌注损伤模型的血清及脑组织中超氧化物歧化酶(SOD)、丙二醛(MDA)、一氧化氮(NO)、一氧化氮合酶(NOS)的含量及病理学改变的影响.方法选用21只沙土鼠并随机分为3组,脑缺血再灌注组(再灌注组)、西比灵预防组(预防组)及对照组,每组7只,分别测定血清、脑组织中SOD、MDA、NO、NOS的含量,并用电镜检测其病理学的变化.结果与对照组比较,再灌注组显示严重的病理损害,其血清、脑组织中MDA、NO、NOS含量明显上升、SOD含量明显下降,两组比较有显著性差异(均P<0.01);与再灌注组比,预防组病理改变较轻,其血清、脑组织中MDA、NO、NOS明显下降,SOD明显上升,两组比较均有显著性差异(分别P<0.01和P<0.05).结论西比灵可减少自由基,诱导SOD生成,具有保护脑细胞的作用.     

线粒体钙单向转运体在大鼠脑缺血再灌注损伤中的作用及其机制

目的观察线粒体钙单向转运体在大鼠脑缺血/再灌注(I/R)损伤中的作用及其机制。方法将40只雄性Wistar大鼠随机分为4组:A组(假手术组)、B组(脑缺血再灌注组)、C组(脑缺血再灌注+钌红组)、D组(脑缺血再灌注+精胺组),采用线栓法建立大鼠大脑中动脉闭塞(MCAO)模型,各组在脑缺血2h后进行再灌注,再灌注24h后观察各组大鼠神经功能评分,检测各组血清脂质过氧化产物丙二醛(malondial-dehyde,MDA)的含量、血清乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)的活性,免疫组化检测皮质区Caspase-3阳性细胞数的变化。结果 B、C、D组神经功能评分升高,血清MDA的含量以及LDH活性显著高于A组,SOD活性与A组相比显著降低,caspases-3表达细胞与A组相比明显增多。C组能明显改善上述结果,而D组相反。结论抑制线粒体钙单向转运体可以明显减轻脑缺血再灌注损伤,其机制可能与减少氧自由基产生、维护线粒体功能有关。     

脑缺血再灌注损伤时 c-fos、c-jun 的表达和细胞凋亡

目的　研究脑缺血再灌注大鼠神经细胞凋亡和原癌基因c fos、c jun表达。 方法　 8周龄健康雄性Wistar大鼠 2 4只 ,随机分为缺血再灌注组、假手术组和对照组 ,每组各 8只。制作大鼠大脑中动脉栓塞 (MCAO)再灌注模型 ,缺血 4h再灌注 2h后断头处死 ,TUNEL法检测神经细胞凋亡 ,免疫组织化学法检测神经细胞c fos、c jun蛋白的表达。结果　缺血再灌注组细胞凋亡率、平均吸光度及c fos、c jun阳性细胞率、平均吸光度均高于假手术组和对照组 (P <0 .0 5 )。结论　脑缺血再灌注损伤可诱导c fos、c jun蛋白的表达和细胞凋亡 ;脑缺血再灌注大鼠神经功能评分与c fos、c jun蛋白的表达和细胞凋亡呈正相关。     

通络救脑注射液对大鼠脑缺血/再灌注神经保护作用的研究

目的观察通络救脑注射液对大鼠脑缺血再灌注损伤的保护作用机制。方法参照Longa法制备大鼠局灶性脑缺血再灌注模型。实验大鼠随机分为假手术组、模型组、通络救脑组。于脑缺血/再灌注24h进行行为学评分,HE染色法检测脑组织病理变化,并且检测大鼠脑组织中超氧化物歧化酶(SOD)、谷胱甘肽(GSH)和丙二醛(MDA)活性的变化,Western blot检测NF-κB的表达结果。结果与模型组相比,通络救脑注射液能明显改善大鼠神经行为,减轻脑组织病理改变,降低MDA含量,明显升高SOD、GSH活性,下调NF-κB表达。结论通络救脑对大鼠脑缺血再灌注损伤神经有保护作用,其作用机制可能为抑制氧化损伤和炎症。     

Effects of transection of cervical sympathetic trunk on cerebral infarct volume and oxygen free radical levels in rats with focal cerebral ischemia/reperfusion injury

BACKGROUND: Stellate ganglion block (SGB) plays a protective role on the brain, but the precise mechanism of action is not clear.OBJECTIVE: To simulate SGB by transection of the cervical sympathetic trunk (TCST) and to investigate the TCST effects on changes in cerebral infarct volume and oxygen free radical levels in rats with focal cerebral ischemia/reperfusion injury.DESIGN, TIME AND SETTING: A complete randomized control animal experiment was performed at the Institute of Neurological Diseases of Taihe Hospital, Yunyang Medical College from February to December 2005.MATERIALS: A total of 101 healthy Wistar rats, weighing 280-320g, of both genders, aged 17-18 weeks, were used in this study. 2,3,5-triphenyltetrazolium chloride (TTC) was purchased from Changsha Hongyuan Biological Company. Superoxide dismutase (SOD), malondialdehyde (MDA) and nitric oxide (NO) assay kits were provided by Nanjing Jiancheng Bioengineering Institute.METHODS: Rats were randomly divided into a TCST group, a model group and a sham operation group. Successful models were included in the final analysis, with at least 20 rats in each group. After TCST, rat models of focal cerebral ischemia/reperfusion injury were established in the TCST group by receiving middle cerebral artery occlusion (MCAO) by the intraluminal suture method for 2 hours, followed by 24 hours of reperfusion. Rat models of focal cerebral ischemia/reperfusion injury were made in the model group. Rats in the sham operation group underwent experimental procedures as for the model group, threading depth of 10mm, and middle cerebral artery was not ligated.MAIN OUTCOME MEASURES: Brain tissue sections of ten rats from each group were used to measure cerebral infarct volume by TTC staining. Brain tissue homogenate of another ten rats from each group was used to detect SOD activities, MDA contents and NO levels. Rat neurological function was assessed by neurobehavioral measures.RESULTS: Cerebral infarct volume was bigger in the model group than in the TCST group (P<0.05). Twenty four hours after cerebral ischemia/reperfusion, SOD activities were lower, whereas MDA contents and NO levels were higher in the TCST and model groups, compared with the sham operation group (P<0.05 or P<0.01). Compared with the model group, SOD activities were higher, whereas MDA contents and NO levels were lower in the TCST group (P<0.05).CONCLUSION: After TCST, cerebral infarct volume is reduced, SOD activities are increased, and MDA contents and NO levels are decreased compared to the model group in rats with focal cerebral ischemia/reperfusion injury. These changes may be associated with TCST.     

β-七叶皂甙钠对脑缺血-再灌注损伤大鼠自由基的影响

目的探讨β-七叶皂甙钠在大鼠脑缺血-再灌注损伤时对自由基的影响。方法用Wistar大鼠45只制备局灶性脑缺血-再灌注模型,分为假手术组、缺血-再灌注组、β-七叶皂甙钠治疗组,每个组又分为缺血2 h后再灌注6 h、12 h、24 h三个时间点,每组每个时间点5只大鼠,术后观察脑组织梗死面积、大鼠的神经行为学变化评分、脑组织超微病理结构变化、脑组织TTC染色,对缺血区脑组织超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondiadehyde,MDA)含量进行测定和分析。结果假手术组相应时间点神经功能损害评分显著低于缺血-再灌注组和治疗组(P<0.05),治疗组SOD含量明显高于缺血-再灌注组各时间点(P<0.05)。光镜显示假手术组脑组织结构未见明显病理改变,脑缺血-再灌注组神经细胞缺血坏死,细胞水肿明显,胶质细胞弥漫增生,炎性细胞浸润。结论自由基参与了脑缺血-再灌注损伤,β-七叶皂甙钠可降低缺血-再灌注后脑组织中MDA的含量,增加SOD的活性,减轻梗死体积,显著减轻大鼠神经功能损害和脑组织水肿,对局灶性脑缺血-再灌注损伤具有一定的保护作用。     

大鼠脑缺血-再灌注损伤中P-选择素的表达对脑血流的影响

目的　探讨脑缺血-再灌注损伤中P-选择素(PS)的表达及其与脑缺血后脑血流恢复的关系。方法　建立脑缺血-再灌注损伤大鼠模型,实验动物随机分为假手术组、非治疗组和藻酸双酯钠治疗组。观察大鼠治疗前后脑组织中PS表达、髓性过氧化物酶(MPO)活性和大鼠脑血流。结果　大鼠脑缺血再灌注损伤组织PS表达及MPO活性增高,缺血侧脑血流量明显减低。藻酸双酯钠治疗组脑组织中PS表达和MPO活性增高受抑,相应的再灌后脑血流量较非治疗组的明显增高。结论　抑制脑缺血-再灌注损伤脑组织中PS表达可改善脑缺血后无复流现象。     

甘草总黄酮对大鼠局灶性脑缺血再灌注损害的脑保护作用

目的　研究甘草总黄酮对大鼠局灶性脑缺血再灌注损害的脑保护作用。方法　我们拟采用线栓法建立的大鼠大脑中动脉 (MCA)缺血再灌注模型 ,并用 3种不同剂量的甘草总黄酮灌服后 ,测定缺血 2h再灌注 2 4h时血清和脑组织中丙二醛 (MDA)、超氧化物歧化酶 (SOD)、一氧化氮 (NO)、一氧化氮合酶 (NOS)活性。结果　发现甘草总黄酮能促进大鼠MCA、缺血再灌注 2 4h后神经功能恢复 ,甘草总黄酮能明显降低血清、脑组织中的MDA、NO含量 ,提高体内SOD的活性。结论　提示中药甘草总黄酮有抗氧化作用     

阿司匹林对沙土鼠全脑缺血-再灌注后脑内一氧化氮合酶及一氧化氮的影响

目的观察阿司匹林对沙土鼠全脑缺血-再灌注后的脑保护作用及其与脑内一氧化氮合酶及一氧化氮水平变化的关系。方法采用夹闭双侧颈总动脉的方法,制备沙土鼠短暂性全脑缺血-再灌注模型。27只健康雄性蒙古沙土鼠随机分为假手术组、脑缺血-再灌注组和阿司匹林治疗组,观察缺血7min再灌注24h后沙土鼠脑组织的病理学改变,以及一氧化氮合酶与一氧化氮水平的变化。结果病理学检查结果显示,沙土鼠脑缺血7min再灌注24h后海马CA1区缺血性损害明显,脑组织内一氧化氮合酶及一氧化氮水平显著升高(P<0.01);与脑缺血-再灌注组相比,阿司匹林治疗组沙土鼠的病理损害较轻,一氧化氮合酶与一氧化氮水平明显下降(P<0.01)。结论阿司匹林可显著减轻脑缺血-再灌注后的脑损伤,其作用机制可能与抑制一氧化氮合酶与一氧化氮水平上升有关。     

莱菔硫烷对大鼠局灶性脑缺血再灌注损伤的保护作用及机制研究

目的 探讨莱菔硫烷对大鼠局灶性脑缺血再灌注损伤的保护作用及机制.方法 采用线栓法制备大鼠大脑中动脉阻断局灶性脑缺血模型,分别于MCAO后1h腹腔注射莱菔硫烷2.5mg/kg、5mg/kg、10mg/kg.于缺血2h再灌注24h时进行神经行为缺损评分,TTC染色评价脑梗死体积,测定脑组织中超氧化物歧化酶(SOD)活力和丙二醛(MDA)含量.免疫荧光组织化学染色法检测黄核蛋白NQ01和脂质过氧化酶Prx6的表达.结果 莱菔硫烷给药组与对照组相比均能改善大鼠脑缺血再灌注后神经行为缺损评分,减少脑梗死体积.其中5mg/kg组能显著改善大鼠脑缺血再灌注后神经行为缺损评分,减少脑梗死体积,增强SOD活性,降低MDA含量.免疫荧光组织化学染色法提示NQ01和Prx6的表达明显增强.结论 莱菔硫烷对大鼠局灶性脑缺血再灌注损伤有神经保护作用,其机制可能与上调内源性抗氧化蛋白NQ01和Prx6的表达有关.     

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。     

尼莫地平对大鼠急性脑缺血再灌注损伤早期保护作用的研究

目的探讨尼莫地平对急性脑缺血再灌注损伤的早期保护作用。方法线栓法复制大鼠急性脑缺血模型。30只雄性Wistar大鼠随机分为假手术、模型、尼莫地平3组。模型组采取缺血2h再灌注2h。尼莫地平组大鼠在缺血0时刻起每小时腹腔注射给药一次,剂量为5mg/kg。各组大鼠在手术4h后实验结束后,行腹主动脉采血,同时取完整脑组织。脑组织切片进行TTC染色,并比较各组脑组织梗死面积。检测各组大鼠血清及脑组织匀浆中SOD、MDA、NO含量。结果与模型组相比,尼莫地平组大鼠脑组织梗死面积显著减少。血清生化指标显示,模型组SOD含量显著低于假手术组,给予尼莫地平治疗后,SOD含量增加明显。模型组MDA、NO含量明显高于假手术组,尼莫地平组明显降低血清中MDA、NO。结论尼莫地平对大鼠急性脑缺血再灌注损伤有保护作用,这种保护作用与NO和氧化应激密切相关。     

Correlation between HSP70 and c-F0S expression and apoptosis in rats undergolng transient focal cerebral ischemla and reperfusion

OBJECTIVE To compare the induction of c-Fos and HSP70 and the presence of apoptosis and the influence of Ginkgo biloba extract (EGb761) in transient focal cerebral ischemic reperfusion rats.BACKGROUND Proto ancogene activation and induction of heat shock protein (HSP) occur in response to cerebral ischemia, but the correlation between these proteins and apoptosis remains uncertain. METHODS Healthy wistar rats were randomized to the normal control group(Group A, n=4), the Sham-operated control group(Group B, n=4), the ischemia and reperfusion group(Group C, n=24), the EGb761 pre-treated ischemia and reperfusion group(Group D, n=24). The rats of Group C and Group D were subjected to transient left middle cerebral artery occlusion (MCAO) as described by Zea longa for 1 hour. RESULTS Immunohistochemical analysis revealed no c-Fos or HSP70-immunoreactivity in Group A and Group B rats. However, in Group C rats, c-Fos was expressed in ipsilateral superficial cortical layers at 1 hour after reperfusion. At 6 hours, c-Fos immunoreactivities were increased in the ipsilateral cortex and were present in the contralateral cortex, while HSP70 were induced beginning in the ipsilateral neurons of MCA distribution. At 12 hours, the expression of c-Fos reached top in superficial cortical layers. At 24 hours HSP70 immunoreactivities reached top both in ipsilateral cortex and in ipsilateral striatum. At 3 days after recirculation, HSP70 expression decreased. c-Fos expression disappeared at day 7 and HSP70 expression only occured in endothelial cells. TUNEL staining showed that there was no cell apoptosis in Gronp A or in Group B. However, in Group C, TUNEL-positive neurons were observed in the border of the penumbra-like area that surrounds the ischemic core at 6 hours following reperfusion and then the number of TUNEL-positive cells reduced gradually. The changes in expression of HSP70 and c-Fos at different time points in Group D was in accord with in Group C, but the number of positive cells and the immunoreactivities in Group D were more intense than in Group C. We found only several TUNEL-positive cells at 6 hours following reperfusion in Group D and no TUNE;L-positive cells were present at other time points.CONCLUSION The present results indicate that c-Fos was expressed from an earlier stage of reperfusion and the expression occured in bilateral cerebral cortex. In contrast, HSP70 induction began later and only occured in ipsilateral neurons of MCA distribution. Our results indicated that EGb761 could increase the expression of c Fos and HSP70 and reduce the apoptosis of neurons.