重复电针预处理对兔脊髓缺血再灌注损伤的保护作用

AIM: To investigate whether pretnatment with repeated electroacupuncture (EA) could induce ischemic tolerance against transient spinal cord ischemia in rabbits. METHODS: 24 male New Zealand white rabbits were randomly assigned to 3 groups(n = 8 each), animals in the control group received no treatment; animals in the SP and EA group received sodium pentobarbitone 30 mg/kg each day for 5 days; animals in EA group were also received electroacupuncture at the Zusanli acupoint 30 min a day for 5 days. 24 hours after the last treatment, spinal cord ischemia was induced by an infrarenal aortic occlusion for 20 min. Hind-limb motor function was determined with the Tarlov criteria at 4,8,12,24 and 48 h after reperfusion. All animals were sacrificed at 48h after reperfusion and the spinal cords(L5) were removed immediately for histopathologic study. RESULTS: The neurologic outcome and histopathology (48 h) in the EA group were significantly better than the control group( P = 0. 006) . CONCLUSION: Pre-ischemic tr     

纳洛酮对重复电针预处理诱导脊髓缺血耐受作用的影响

目的 :研究阿片肽在电针重复预处理诱导脊髓缺血耐受中的作用。方法 :4 0只雄性新西兰大白兔随机分成 5组 (n=8) :对照组、戊巴比妥钠组、纳洛酮组、电针预处理组和纳洛酮 +电针预处理组。对照组未行任何处理 ;戊巴比妥钠组每日静脉给予戊巴比妥钠 30 m g/ kg,连续 5 d;纳洛酮组每日静脉给予纳洛酮 0 .3mg/ kg,连续 5 d;电针预处理组每日在戊巴比妥钠 30 m g/ kg麻醉下 ,电针刺激双侧委中穴 6 0 min/ d,连续 5 d;纳洛酮 +电针预处理组预处理前 30 m in静脉注射纳洛酮 0 .3mg/ kg,余同预处理组。最后一次预处理后 2 4 h阻闭肾下腹主动脉 2 0 min,制备兔脊髓缺血模型 ;再灌注后 4 h、8h、12 h、2 4 h和 4 8h分别对动物后肢运动功能进行评分 ;再灌注 4 8h后深麻醉下处死动物取脊髓 (L 5～ L 7) ,制作标本并行组织病理学观察。结果 :所有动物都存活 ,再灌注 4 8h后电针预处理组动物后肢运动功能评分及脊髓前角运动神经元计数均明显高于纳洛酮 +电针预处理组 (P均 <0 .0 1) ,对照组、戊巴比妥钠组、纳洛酮组及纳洛酮 +电针预处理组间后肢运动功能评分及脊髓前角运动神经元计数均无显著性差异 (P均 >0 .0 5 )。结论 :重复电针预处理对兔脊髓缺血再灌注损伤的保护作用可被纳洛酮阻断 ,提示阿片肽参与了电针预处     

不同穴位重复电针预处理诱导兔脊髓缺血耐受效果的对比

目的:比较不同穴位电针预处理对兔脊髓缺血再灌注损伤保护作用的效果,为临床提供最佳的预处理方案。方法:32只雄性新西兰大白兔随机数字表法分成4组(各组n=8),即对照组、戊巴比妥钠组、委中穴组及足三里穴组。对照组静脉给予生理盐水1mL/kg,连续5d;戊巴比妥钠组静脉给予戊巴比妥钠30mg/kg,连续5d;委中穴和足三里穴组每天在戊巴比妥钠(30mg/kg)麻醉下,电针分别刺激委中穴和足三里穴60min/d,连续5d。最后一次预处理后24h,夹闭肾下腹主动脉20min,制作兔脊髓缺血模型;再灌注后4,8,12,24和48h分别对动物后肢运动功能评分;再灌注48h后,深麻醉下处死动物取脊髓(L5～7),制作标本行组织病理学观察。结果:所有动物均存活,再灌注后48h电针预处理委中穴和足三里穴组后肢运动功能评分及脊髓前角运动神经元计数均明显高于对照组(P=0.001);足三里组后肢运动功能评分和脊髓前角运动神经元计数明显低于委中穴组(P=0.001);对照组与戊巴比妥钠组相比,后肢运动功能评分及脊髓前角神经元计数无显著性差别(P=1.0和P=0.873)。结论:电针预处理对兔脊髓缺血再灌注损伤有显著的保护作用,且刺激委中穴优于刺激足三里穴的效果。     

远程预处理对兔脊髓缺血再灌注损伤的保护作用(英文)

目的:探讨远程预处理对兔脊髓缺血再灌注损伤的保护作用。方法:20只成年雄性新西兰大白兔随机分成2组(各组n=10),即对照组和远程预处理(RPC)组。所有动物脊髓缺血前1h戊巴比妥钠麻醉动物(30mg/kg,iv)。RPC组动物麻醉后进行双后肢短暂缺血2次(充气式压力止血带环扎双后肢,压力26.6kPa,阻断10min,松开10min,再阻断10min),最后一次缺血后再灌注30min,阻闭肾下腹主动脉20min,制作兔脊髓缺血模型。对照组动物不进行双后肢短暂缺血,其余同RPC组。再灌注后4,8,12,24和48h分别对动物后肢运动功能评分。再灌注48h后,处死动物取脊髓(L5～7),石蜡包埋切片行组织病理学观察。结果:RPC组神经功能评分在各时间点均明显高于对照组(P=0.000);与对照组相比,RPC组脊髓前角正常神经细胞数明显增多(P=0.000),且神经功能评分与脊髓前角正常神经细胞计数之间有显著相关性(r=0.868,P<0.01)。结论:远程预处理对兔脊髓缺血再灌注损伤有显著的保护作用。     

硫酸镁对兔脊髓缺血再灌注损伤的保护

背景硫酸镁用于脑缺血再灌注损伤的治疗已取得了较满意的疗效,但对脊髓缺血再灌注损伤的作用机制尚不十分清楚.目的观察硫酸镁对兔脊髓缺血再灌注损伤的保护效果,进一步探讨其作用机制.设计以实验动物为研究对象,随机对照的重复测量设计.单位一所大学医学院中心实验室.对象实验于2003-04/2004-06在西安交通大学医学院中心实验室完成.健康成年新西兰大白兔27只,体质量1.9～2.5 kg.随机抽签法分为硫酸镁组、生理盐水组和假手术组,每组9只.方法夹闭腹主动脉肾下段30min后恢复血流再灌注48 h,建立兔脊髓腰骶段缺血模型.硫酸镁组给予静脉灌注硫酸镁(0.25 mL/kg·h),生理盐水组用等量生理盐水代替.假手术组仅行中线剖腹术,不结扎动脉.在缺血前,缺血30 min及再灌注后1,2,8,16,24 h对动物行体感诱发电位监测,再灌注24及48 h后对硫酸镁组、生理盐水组动物行运动功能评分.再灌注后48 h处死动物,对脊髓行组织病理学检查.主要观察指标①运动功能评分.②体感诱发电位监测.③脊髓组织病理学检查.结果缺血30 min时硫酸镁组体感诱发电位(N1)潜伏期明显延长;再灌注后前2 h潜伏期较缺血时明显恢复,其后又显著延长.缺血30 min时生理盐水组波形消失.假手术组体感诱发电位没有明显变化,动物均完全康复.硫酸镁组各时间点潜伏期恢复均明显高于生理盐水组(P＜0.05);再灌注24和48 h后,硫酸镁组的神经功能评分分别为(3.7±0.5)和(3.4±0.7)分,均显著高于生理盐水组[(3.0±0.7)和(2.6±0.9)分](P＜0.05);再灌注48 h后硫酸镁组的脊髓前角正常神经细胞计数显著高于生理盐水组(23.4±3.4,12.3±3.2,P＜0.01).结论硫酸镁具有减轻兔脊髓缺血再灌注损伤及保护神经功能的作用.     

电针对心肌缺血再灌注损伤家兔心肌细胞凋亡的影响

目的:研究电针对心肌缺血再灌注损伤家兔心肌细胞凋亡的影响.方法:将结扎左冠状动脉前降支30 min,再灌注60 min,成功制作为心肌缺血再灌注损伤模型的家兔24只,随机分为模型组、电针1及2组各8只.另仅开胸未结扎冠状动脉的8只为假手术组作为正常对照.电针1、2组术后30 min分别采用电针针刺内关穴与列缺穴60 min,仅1次,应用原位末端标记法观察4组家兔心肌细胞凋亡情况.结果:心肌凋亡细胞数量,模型组显著高于假手术组(P＜0.01),电针1组与电针2组、模型组比较显著降低(P＜0.05,P＜0.01),但仍高于假手术组(P＜0.01).结论:细胞凋亡参与心肌缺血再灌注损伤的病理过程,电针内关穴可以减少缺血再灌注损伤心肌细胞凋亡的数量,从而达到抗缺血再灌注损伤的作用.     

重复高压氧预处理对兔短暂脊髓缺血后线粒体结构及ATP酶活性的影响

背景重复高压氧预处理诱导脊髓对随后的缺血产生耐受,这一现象的证实为临床防治胸腹主动脉术后的脊髓缺血并发症开辟了新的道路.探讨其缺血耐受性产生的机制无疑将为这一方法的临床应用奠定基础.目的探讨重复高压氧预处理诱导脊髓缺血耐受中线粒体结构及ATP酶活性变化.设计随机对照实验. 单位解放军第四军医大学西京医院麻醉科.材料实验于2004-03/06在第四军医大学空医系高压氧舱及西京医院麻醉科实验室完成.选用雄性新西兰大白兔50只.方法50只雄性新西兰大白兔随机分为两组单纯缺血对照组,重复高压氧预处理组0.25 MPa,100%O2,1 h/d,5 d.每组25只.最后1次预处理后24 h制作脊髓缺血模型.采用肾下腹主动脉阻断法造成脊髓缺血(20 min)/再灌注模型,观察动物再灌注6,24和48 h后肢神经运动功能评分,分别于缺血前、再灌注6,24和48 h处死动物并提取线粒体测定ATP酶活性.主要观察指标①各组动物后肢运动神经功能评分.②各组动物脊髓线粒体ATP酶活性.③各组动物病理学评估.结果纳入分析50只,最终进入统计分析的大白兔保持为2组,各25只,无缺失值.①后肢运动神经功能评分再灌注6,24和48 h高压氧预处理组神经功能评分明显高于对照组(P＜0.01).②脊髓线粒体ATP酶活性两组动物再灌注后各时间点线粒体Na+,K+-ATP酶活性和Ca2+,Mg2+-ATP酶活性均明显下降,高压氧预处理组Na+,K+-ATP酶活性在再灌注6,24h和48 h明显高于对照组(P＜0.01);Ca2+,Mg2+-ATP酶活性在再灌注6 h和24 h明显高于对照组(P＜0.01).③病理学评估电镜下观察再灌注48 h时的组织切片,对照组线粒体空泡化明显,高压氧预处理组多数线粒体结构接近正常.结论高压氧预处理诱导脊髓缺血耐受产生的机制部分可能是通过减慢线粒体ATP酶活性下降,维持线粒体功能实现的.     

重组人促红细胞生成素对兔脊髓缺血/再灌注损伤后白细胞介素8表达的影响

目的探讨重组人促红细胞生成素(rHuEPO)对兔脊髓神经细胞缺血/再灌注损伤后白细胞介素8(IL-8)表达的影响。方法 44只健康成年新西兰大白兔随机分为3组:假手术组(4只),对照组(20只),rHuEPO处理组(20只),对照组与rHuEPO处理组分别于再灌注6 h,12 h,24 h、48 h、7 d后时间点随机处死4只动物,取L3～L5节段脊髓组织用霉菌抗生物素蛋白-过氧化酶免疫组化染色法(S-P法)及兔抗人IL-8 ELISA试剂盒检测脊髓组织内IL-8的表达。结果 IL-8在无损伤脊髓中即见有表达。随着缺血及再灌注进程的加重,组织中IL-8阳性表达强度逐渐增加。脊髓损伤后24 h表达明显上调并达高峰;高表达持续至损伤后7 d;两组间IL-8水平的变化趋势相同,但在各时间点rHuEPO处理组兔脊髓组织IL-8的含量均低于对照组(P<0.05)。结论 rHuEPO具有抑制脊髓组织炎症反应的作用,能明显抑制兔脊髓缺血/再灌注损伤后的IL-8表达。     

Protective effects of trimetazidine in transient spinal cord ischemia

The neuroprotective effect of trimetazidine (TMZ) was tested prospectively in a rabbit spinal cord ischemia model. Ischemia was induced by clamping the aorta just distal to the left renal artery and proximal aortic bifurcation for 20 min. Twenty-five male New Zealand white rabbits were randomized as follows: TMZ group (n=10) receiving 3 mg/kg trimetazidine intravenously before the occlusion of the aorta; control group undergoing occlusion but receiving no pharmacologic intervention (n=10); sham-operation group (n=5) subjected to operative dissections without aortic occlusion. Physiological parameters and somatosensory evoked potentials (SEP) were monitored in animals before the ischemia, during the ischemia and in the 1st, 15th and 60th min of reperfusion. Neurologic status was assessed 24 and 48 h after the operation. The spinal cord, abdominal aorta, and its branches were processed for histopathologic examinations 48 h after the operation. At the end of the ischemic period, the average N1-P1 amplitude was reduced to 22% of the baseline in all ischemic animals. This was followed by a gradual return to 90DŽ% of the initial amplitude in the TMZ group and 81DŽ% in the control group (P<0.05) after 60 min of reperfusion. The average motor function score was significantly higher in the TMZ group than the control group (3.7ǂ.5 vs 3.1ǂ.6 at 24 and 3.5ǂ.7 vs 2.9ǂ.6 at 48 h; P<0.05). Histologic observations were clearly correlated with the neurologic findings. The results suggest that trimetazidine reduces spinal cord injury during thoracoabdominal aortic operations and may have therapeutic utility during high risk operations.     

氨基胍对兔脊髓缺血再灌注损伤的保护作用

目的：研究氨基胍对兔脊髓缺血再灌注损伤的保护作用。方法：在左肾动脉下方夹闭腹主动脉30min后恢复血流灌注,建立兔脊髓腰骶段缺血再灌注损伤模型。18只新西兰大白兔随机分为假手术组（A组,n=4）、缺血再灌注组（B组,n=7）和氨基胍治疗组（C组,n=7）。C组在夹闭动脉前静脉注射氨基胍（100mg.kg-1）,术后8h再肌肉注射氨基胍（100mg.kg-1）1次。术后24h对兔后肢运动功能进行评分,观察脊髓病理改变情况,检测脊髓一氧化氮（NO）含量和一氧化氮合酶（NOS）活性,TUNEL法检测脊髓细胞凋亡,免疫组织化学染色测定Bax与Bcl-2蛋白表达情况。结果：缺血再灌注后24h,与缺血再灌注组相比,氨基胍治疗组兔后肢运动功能明显改善,脊髓内NO含量和NOS活性下降（P〈0.05）,脊髓细胞凋亡指数下降（P〈0.05）,Bax蛋白表达下降（P〈0.05）,Bcl-2蛋白表达升高（P〈0.05）,脊髓损伤病理改变明显减轻。结论：氨基胍对兔脊髓缺血再灌注损伤有保护作用,其机制与抑制iNOS活性有关,并且可以通过调节Bax、Bcl-2蛋白表达抑制脊髓细胞凋亡。     

电针刺复合丹参对缺血再灌注心肌的协同保护作用

背景电针刺和丹参均具有防治缺血再灌注损伤的作用,但当两者同用时是否可产生协同作用. 目的探讨电针剌和丹参对缺血再灌注后具有心肌保护作用的心肌细胞热休克蛋白70 mRNA表达以及心肌损伤的标志多巴胺水平的影响,以及电针刺和丹参之间的相互作用. 设计两因素析因设计. 单位上海第二医科大学附属仁济医院麻醉科. 材料实验于2001-09/2002-12在仁济医院动物实验室进行.取康新西兰白兔24只,随机分为4组,每组6只①缺血再灌注组.②电针组.③丹参组.④电针+丹参组. 方法①缺血再灌注组采用夹闭心脏冠状动脉前降支30 min后松解2 h制成缺血再灌注模型.②电针组同前造模,在缺血前20 min电针刺"内关","云门","列缺",电压0.8 V,频率3.0～4.0 Hz.③丹参组同前造模,缺血前静脉单次注射丹参1.5 mg/kg,灌注前和灌注后分别单次注射丹参1.0 mg/kg.④电针剌+丹参组同前造模,既电针又给药.在缺血前,缺血30 min和再灌注2 h分别取各组兔静脉血测定血中多巴胺的含量;取缺血区和非缺血区心肌热休克蛋白70 mRNA的表达通过反转录-聚合酶链反应方法测定.结果经补充后24只兔进入结果分析.①心肌热休克蛋白70 mRNA的表达各组缺血再灌注后均较缺血前显著增加(P＜0.01),其他3组均高于缺血再灌注组(P＜0.05),电针+丹参组高于电针组和丹参组(F=4.48,P＜0.05).②多巴胺水平各组缺血再灌注后均较缺血前显著升高(P＜0.01),其他3组缺血30 min和再灌注2 h时血中多巴胺水平显著低于缺血再灌注组(P＜0.05),电针+丹参组低于电针组和丹参组(F=5.95,P＜0.05). 结论①电针刺和丹参都能激发缺血再灌注后热休克蛋白70 mRNA的表达,增强热休克蛋白70的蛋白稳定作用,减轻心肌损害.②电针剌和丹参能抑制缺血再灌注后体内多巴胺含量的增加,,减少多巴胺介导的损伤,保护心肌.③电针刺和丹参之间有相互协同效应.     

Hyperbaric oxygen therapy given 30 minutes after spinal cord ischemia attenuates selective motor neuron death in rabbits

OBJECTIVE: Spinal cord ischemia sometimes causes paraplegia because the spinal motor neuron cells are vulnerable to ischemia. Although various protective remedies for spinal cord injury have been reported, there have been few established clinical methods. Although hyperbaric oxygen (HBO) has been used clinically as a treatment for ischemia, the reason for its effectiveness is still uncertain because sufficient experimental data are lacking. DESIGN: Prospective, randomized, controlled study. SETTING: Experimental animal research laboratory in a university research center. SUBJECTS: Twenty-three Japanese white rabbits, weighing 2-3 kg. INTERVENTIONS: A modified rabbit spinal cord ischemia model of infrarenal aortic occlusion for 15 mins was employed. Rabbits were randomly assigned to four groups; the rabbits in group A did not undergo ischemic insults (n = 5). The rabbits in groups B and C underwent ischemic insult for 15 mins, followed by 1 hr of HBO treatment at 3 atm absolute with 100% oxygen at 30 mins (n = 6) or 6 hrs (n = 7) after reperfusion, respectively. The rabbits in group D underwent ischemic insult for 15 mins without HBO treatment (n = 5). MEASUREMENTS AND MAIN RESULTS: We observed neurologic functions for 14 days. The sections of the spinal cords were stained with hematoxylin and eosin, and the number of spinal motor neurons in ventral region was counted by light microscopy. All rabbits in groups A and B could stand, whereas all rabbits in groups C and D showed irreversible paraplegia on days 2 and 14 after reperfusion. Spinal motor neurons in ventral gray matter in groups C and D decreased significantly compared with those in groups A and B. CONCLUSIONS: HBO therapy shortly after ischemic insult had protective effects against ischemic spinal cord damage. However, delayed treatment with HBO did not change the prognosis.     

电针对MCAO大鼠脑皮质EPO表达和脑血流量的影响

目的：观察电针疗法对局灶性脑缺血大鼠脑皮层EPO表达和缺血局部组织血流量的影响,探讨其作用机制.方法：SD大鼠80只,随机分为正常组和假手术组各10只、模型组和电针组各30只;模型组和电针组又各按缺血再灌注24h、48h和72h分别分为3个亚组,每组10只.假手术组仅分离血管而不插线栓,模型组和电针组制备大脑中动脉缺血再灌注模型（MCAO）,电针组采用电针治疗.采用免疫组化组法及免疫印迹法检测大鼠脑皮质中促红细胞生成素（EPO）蛋白的表达,用激光多谱勒血流仪检测大鼠脑组织血流量（rCBF）的变化.结果：EPO免疫组化及Western结果比较,模型组及电针组EPO阳性细胞及蛋白表达均在脑缺血再灌注24h开始增加,48h达到峰值,72h开始下降,且电针组在3个时间点均高于同时间点模型组（P＜0.05,0.01）.rCBF比较,模型组和电针组随着24、48、72h 3个时间点呈明显增加趋势,且电针组各时间点rCBF均较模型组明显升高（P＜0.05,0.01）.结论：电针能使脑缺血再灌注脑组织中EPO表达增加,明显提高局部脑组织血流量,有助于减轻脑缺血再灌注后脑组织损伤.     

阿托伐他汀预处理对兔脊髓缺血再灌注损伤的影响

目的研究不同剂量阿托伐他汀预处理后,家兔脊髓缺血再灌注损伤模型行为学及生化学改变,评价阿托伐他汀预处理对脊髓缺血再灌注损伤的影响。方法用腹主动脉夹闭法制备脊髓缺血再灌注模型,缺血45 min,分别于动物再灌注2 h、6 h、12 h、24 h行动物行为学评分;再灌注24 h后,截取脊髓组织。检测脊髓组织匀浆中的丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性及髓过氧化物酶(MPO)活性。结果实验动物缺血再灌注12 h、24 h后行为学评分明显下降,中高剂量阿托伐他汀能明显增强SOD活性、降低MDA含量及MPO活性。结论阿托伐他汀预处理对脊髓缺血再灌注损伤有一定的神经保护作用,其疗效与剂量有关。     

多次缺血预处理对兔脊髓缺血性损伤时金属元素的影响

背景缺血预处理对主动脉手术的脊髓缺血性损害有良好的保护作用,但是脊髓缺血预处理保护作用的机制尚未完全阐明.目的探讨多次缺血预处理对兔脊髓缺血再灌注损伤的保护作用及其机制.设计以实验动物为研究对象,完全随机对照实验研究.单位一所大学医院的麻醉科.材料实验于2002-09/12在武汉大学人民医院麻醉学研究室完成.24只日本大白兔随机双盲分为假手术组、缺血再灌注组和缺血预处理保护组,每组8只.干预假手术组不阻断主动脉,缺血再灌注组阻断主动脉45 min,缺血预处理保护组阻断主动脉5 min,开放5 min,反复4次之后再阻断45 min.主要观察指标术后第7天检测脊髓组织金属元素(钙,镁,铜,锌)的浓度.术后观察后肢神经功能的评分、后肢针电极肌电图和脊髓组织病理学的改变.结果缺血再灌注组脊髓组织钙,铜的浓度较假手术组显著性升高(P＜0.05或0.01),镁,锌的浓度则显著性降低(P＜0.05).缺血再灌注组脊髓组织钙、锌的浓度分别较缺血预处理保护组显著性升高或降低(P＜0.01).缺血再灌注组后肢神经功能评分均显著性低于假手术组和缺血预处理保护组(P＜0.05或0.01),脊髓病理学和后肢肌电图亦较缺血预处理保护组有显著性病理改变(P＜0.01).结论多次缺血预处理对兔脊髓缺血再灌注损伤具有显著而又快速的保护作用,其保护机制与维持缺血区域钙,镁,铜,锌离子的平衡有关.     

脊髓损伤后孕酮保护作用研究

目的:探讨脊髓损伤后孕酮的保护作用。方法:将54只日本大耳白兔随机均分为假手术组、脊髓损伤组与孕酮治疗组各18只,每组又分为治疗后12 h、24 h、36 h、48 h、72 h和14 d 6个时间点。假手术组只行腹部切开术但不阻断腹主动脉,脊髓损伤组和孕酮治疗组制备缺血再灌注模型,造模后孕酮治疗组注射孕酮治疗,脊髓损伤组在相同时间点注射等量生理盐水。比较各时间点不同组兔神经细胞超微结构变化情况和神经功能(BBB)评分。结果:电镜结果显示,脊髓损伤组可见神经细胞核固缩,异染色质浓染凝集成块,细胞浆中有大量空泡形成。孕酮治疗组可见神经细胞和胶质细胞的核轻度固缩,染色质浓染、细胞浆空泡化均较脊髓损伤组减轻。脊髓损伤组术后12 h BBB评分为0.6分,随后缓慢升高,术后14 d时评分为9.3分。孕酮治疗组BBB评分变化与脊髓损伤组相似,但术后36 h开始,各时间点评分均高于相应脊髓损伤组(P0.05)。结论:脊髓损伤后,采用孕酮治疗可减轻神经细胞变性,发挥神经保护作用。     

电针合谷穴对大鼠局灶性脑缺血再灌注后脑内毛细血管密度及CD34表达的影响

目的观测电针大鼠双侧合谷穴(LI04)对局灶性脑缺血再灌注后脑内CD34表达及血管新生的影响。方法90 只大鼠随机分为正常组(n=6)、模型组(n=42)和电针组(n=42)。模型组和电针组采用线栓法制备大鼠局灶性脑缺血再灌注模型,电针组采用电针治疗。局灶性脑缺血1 h 后再灌注,观察再灌注后2 h、12 h、24 h、3 d、7 d、14 d、21 d 共7 个时相点大鼠神经症状学评分,采用HE染色观察并计算梗死体积和毛细血管密度,免疫组织化学法检测CD34的表达。结果电针组与模型组比较,再灌注后各时间点神经症状学评分降低(P<0.05)。电针组梗死体积显著小于模型组(P<0.001)。与模型组相比,电针组毛细血管密度再灌注后2 h 开始增加(P<0.05),再灌注后24 h 到再灌注后14 d 均较模型组明显增加(P<0.01)。电针组再灌注后3 d CD34阳性细胞表达较模型组明显增加(P<0.01),7 d 增加达高峰(P<0.01),14 d 仍有显著性差异(P<0.05)。结论电针能促进局灶性脑缺血再灌注后CD34的表达,增加毛细血管密度,促进血管新生。     

电针对神经病理性疼痛大鼠痛觉过敏及脊髓背角钾-氯离子协同转运体2表达的影响

目的:观察患侧电针和健侧电针足三里穴、阳陵泉穴对神经病理性疼痛大鼠行为学表现以及脊髓背角钾-氯离子协同转运体2(KCC2)表达的影响,探讨患侧选穴和健侧选穴两种针刺方案的镇痛效应,以及KCC2在电针镇痛效应中的作用。方法:将40只雄性SD大鼠随机分为4组:假模组、模型组、患侧电针组、健侧电针组,每组10只。建立大鼠坐骨神经慢性压迫性损伤(CCI)模型,治疗组于术后1周开始电针治疗,每天1次,连续7天。各组于术前(0天)及术后3、5、7、10、12、14天分别测量大鼠患侧足机械足反射阈值(MWT)和热缩足反射潜伏期(TWL)。术后14天处死大鼠,取脊髓组织行HE染色观察组织病理学改变,并采用免疫组化方法检测脊髓背角KCC2蛋白的表达。结果:与术前比较,CCI各组大鼠痛阈明显降低,出现痛觉过敏(P0.001),电针治疗后大鼠MWT和TWL均有不同程度的持续升高,且患侧与健侧电针相比无明显差异。HE染色结果提示,患侧电针组与健侧电针组脊髓背角组织及神经元病变程度较模型组减轻。术后14天患侧与健侧电针组较模型组脊髓背角患侧KCC2表达显著增加(P0.05),并且患侧电针组与健侧电针组无明显差异。结论:患侧电针和健侧电针能产生类似的镇痛效果,均可减轻外周神经损伤后引起的痛觉过敏。患侧电针与健侧电针治疗均可抑制KCC2蛋白的表达下调,电针的镇痛作用可能是通过增加脊髓背角中KCC2的表达实现的。     

脊髓缺血再灌注损伤中热休克蛋白表达的研究

目的研究脊髓缺血再灌注损伤(SCII)后热休克蛋白(HSP70)表达的变化。方法制作SCII动物模型,采用光镜、激光多普勒超声、免疫组化等技术,研究SCII后HSP70表达的变化。结果(1)缺血30min,血灌流量平均下降85.37%,再灌流即刻血灌流量迅速升高,再灌流10min左右达到最高点。再灌流30min,血灌流量基本恢复到缺血前的基线水平,以后逐渐降低。(2)缺血后部分大鼠再灌注后出现“二次瘫痪”现象。(3)脊髓缺血30min再灌注60min后即有HSP70的表达增强,随着再灌注时间的延长,HSP70表达逐渐增多,在再灌注240min达到高峰。此后,HSP70表达逐渐减少,在再灌注24h基本消失。结论(1)脊髓缺血一定时间后恢复血液供应,可以发生再灌注损伤。(2)脊髓缺血再灌注后HSP70的表达增加。     

Neuroprotective effects of progesterone in spinal cord ischemia in rabbits

BackgroundThis experimental study was performed to investigate the neuroprotective effects of progesterone on spinal cord ischemia in rabbits.MethodsEighteen female New Zealand white rabbits were used in this study. Rabbits were randomized into 3 groups. Spinal cord ischemia was induced by clamping the abdominal aorta from a point just inferior to the left renal artery to the aortic bifurcation for a period of 30 minutes. Group 1 served as the control group, and groups 2 and 3 received intraperitoneal progesterone immediately after the onset of reperfusion, at a dose of 8 mg/kg. Two hours after reperfusion, the animals in group 1 were killed. Four hours after reperfusion, the animals in group 2 were killed, and 6 hours after reperfusion, the group 3 rabbits were killed. Spinal cords were removed and fixed in 10% formalin in a phosphate buffer. Neuronal injury was evaluated by a pathologist who was blinded to the treatment groups, and 5 sections per animal were evaluated. The number of intact large motor neuron cells in the ventral grey matter region was counted.ResultsThe analysis revealed that the average mean arterial pressure for group 1 was significantly higher than that for group 2, and the mean sacrificed pressure value for group 1 was significantly higher than that for group 3 (P < .05). The number of intact neurons in group 1 was significantly lower than the number of intact neurons found in both groups 2 and 3 (P < .05). No other statistically significant differences were found between the groups.ConclusionThe findings from the present study indicate that progesterone effectively protects the spinal cord tissues against ischemic damage in the setting of decreased perfusion.