Cell proliferation and nestin expression in cortex and SEZ of MCAO rats

AIM:To explore the effect of brain ischemia injury on cell proliferation and nestin expression in cortex and subependymal zone (SEZ).METHODS:Using a local brain ischemia model(MCAO), BrdU positive cells of cortex and subependymal zone (SEZ), also nestin positive cells, were observed by immunohistochemistry.RESULTS:BrdU and nestin positive cells in SEZ of MCAO rats were obviously increased. In cortex, only nestin positive cells were observed.CONCLUSION:Neural stem cells in SEZ and cortex were activated after brain ischemia, it may be related with neural recovery after brain ischemia injury.     

A reactive pattern of ischemia-induced nestin protein in the rat brain

AIM:To explore the expressive profile of nestin protein in the focal ischemic brain and to study the recovery mechanism of brain focal infarct.METHODS:Cellular morphology,time-course and distribution pattern of nestin positive response were immunohistochemically examined in different brain regions of 36 adult male SD rats. RESULTS:Nestin positive response of different brain regions in sham operated rats was present in small- and micro-vasculartures and the third ventricle bottom and ependyma. A large number of nestin positive cells were detected in ischemic brain, and were more remarkable in the cortical areas of parietal lobe and preoptic area as well as ischemic caudoputamen. Stellate nestin positive cells were located in the deep layer of ischemic cortex, but fibrillary cells were located in the shallow layer. Nestin positive cells in the ischemic caudoputamen showed the same changes of morphology as those cells in the deep layer of ischemic cortex. Morphological and number alterations of nestin positive cells were the most remarkable at 1 weeks post-ischemia, which showed more hypertrophy and proliferation in morphology, and a marked increase in number was present in the ischemic cerebral cortex and the ischemic caudoputamen. These alterations of nestin positive cells persisted up to 6 weeks post-ischemia, and then, the nestin positive response in the ischemic brain decreased gradually.CONCLUSION:Focal cerebral ischemia induces nestin re-expression on reactive astrocytes, which may be very important to the self-recovery of cerebral infarct.     

Astrocyte proliferation in the rat hippocampus after middle cerebral artery occlusion

AIM: To study rat astrocyte proliferation in ipsilateral hippocampus following focal cerebral ischemia. METHODS: Ischemia was induced by temporary middle cerebral artery occlusion (MCAO). In hippocampus of rats at 3, 7 and 30 days after MCAO, the numbers and anatomic distribution of glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry. The protein expression of GFAP and proliferating cell nuclear antigen (PCNA) in the ipsilateral hippocampus were analyzed by Western blot analysis. RESULTS: Astrocytes appeared hypertrophic, with increased process thickness and numbers at 7 days after MCAO, and the highest density of astrocytes were seen at 30 days in the CA1, CA2 regions of the ipsilateral hippocampus. Western blot analysis revealed that GFAP levels were normal at 3 days, but increased by 7 days and remained elevation at 30 days. Western blot analysis of PCNA protein also revealed identified upregulation PCNA at 3 days after MCAO and the expression peaked at 7 days. CONCLUSION: This study demonstrates that focal cerebral ischemia in the rat results in a rapid response, a process often referred to as reactive astrogliosis or glial scarring, from resident astrocytes of the ipsilateral hippocampus to the side of ischemia.     

Curcumin attenuates rat lung ischemia/reperfusion injury by interfering with autophagy

AIM To investigate the role of curcumin （CUR） in lung ischemia/reperfusion （I/R） injury （LIRI） and its relationship with autophagy. METHODS 40 SD rats were randomly divided into sham operation （sham） group, I/R group, solvent （DMSO） group, CUR group and CUR+rapamycin （CUR-Rap） group. The rats were intraperitoneally injected with normal saline, DMSO, CUR or CUR+Rap before operation. After the rat LIRI model was established, the lung tissues were taken to measure W/D, TLW, IAR, and the contents of SOD and MDA were also measured to indicate the oxidative stress level. Light and electron microscopes were used to observed the morphology and ultrastrucure of lung tissues. The expression levels of autophagy-related proteins were determined by Western blot to evaluate autophagy levels. RESULTS Compared with sham group, wet weight/dry weight （W/D）, total lung water （TLW）, injured alveoli rate（IAR） and malondialdehyde （MDA） content in all other groups were increased, superoxide dismutase （SOD） activity was decreased, the levels of autophagy were increased （P<0.05）, and lung tissue injury and cell ultrastructural damage were aggravated in CUR group. Compared with DMSO group, W/D, TLW and IAR and MDA content were decreased, SOD activity was decreased, autophagy levels were also decreased （P<0.05）, and lung tissue and cell ultrastructural damage were attenuated. Compared with CUR group, W/D, TLW, IAR and MDA content were increased, SOD activity declined, the autophagy levels were increased （P<0.05）, and damage of lung tissues and cells were more serious in CUR-Rap group. CONCLUSION Curcumin attenuates the lung I/R injury in rats, and its mechanism may be related to the reduction of oxidative stress and the inhibition of autophagy.     

白肉灵芝水提物对脑缺血大鼠海马神经元的保护作用

试验旨在探讨白肉灵芝水提物（Ganoderma leucocontextum aqueous extracts,GLAE）对脑缺血后海马神经元的保护作用及机制。将50只健康大鼠分为对照组、模型组、GLAE低（0.05 mg/（g·BW））、中（0.1 mg/（g·BW））、高（0.2 mg/（g·BW））剂量组。利用双侧颈总动脉夹闭法建立大鼠脑缺血模型,GLAE组灌胃不同剂量的GLAE干预,对照组和模型组灌胃同体积的生理盐水,连续2周。用跳台试验方法检测记忆获得、记忆巩固和记忆再现障碍大鼠的学习记忆能力,HE染色观察大鼠海马组织的病理形态的变化,比色法检测海马组织一氧化氮合酶（nitric oxide synthase,NOS）活性和一氧化氮（nitric oxide,NO）含量,Western blotting和实时荧光定量PCR法分别检测海马组织生长相关蛋白-43（growth associated protein-43,GAP-43）和脑源性神经生长因子（brain derived neurotrophic factor,BDNF）的水平。结果显示,与对照组相比,模型组大鼠跳台试验的逃避潜伏期显著缩短、电击次数显著增加（P<0.05）;海马神经元细胞出现明显核固缩、排列松散紊乱等退行性改变,细胞数量显著减少（P<0.05）;海马组织NOS活性和NO含量均显著降低（P<0.05）;大鼠海马组织GAP-43蛋白表达量显著升高（P<0.05）;海马组织BDNF mRNA表达量显著下调（P<0.05）。与模型组相比,GLAE干预后,大鼠逃避潜伏期均显著延长、电击次数均显著减少（P<0.05）;GLAE高剂量组大鼠CA1区和齿状回锥体神经元细胞形态明显改善,神经元数量显著增加（P<0.05）;GLAE低剂量组对NOS活性影响不明显（P>0.05）,显著增加NO含量（P<0.05）,GLAE中、高剂量组NOS活性和NO含量均显著升高（P<0.05）;GLAE低、中、高剂量组海马组织GAP-43蛋白表达量均显著增加（P<0.05）;GLAE低、中、高剂量组海马组织BDNF mRNA表达量均显著增加（P<0.05）。以上结果表明,GLAE可通过提高NOS活性和NO水平、促进海马神经发生和功能恢复对脑缺血后海马神经元损伤有一定的保护作用,从而改善大鼠认知功能,0.2 mg/g GLAE效果最好。     

Effects of capsaicin on cognitive impairment and mitochondria-associated endoplasmic reticulum membranes in rats with chronic cerebral hypoperfusion

AIM:To observe the effects of capsaicin on cognitive impairment and mitochondria-associated endoplasmic reticulum membranes (MAMs) of hippocampal CA1 area in the rats with chronic cerebral hypoperfusion (CCH), and to investigate the underlying molecule mechanism of cognitive defects induced by ischemia. METHODS:Healthy male Sprague-Dawley (SD) rats(n=48) were randomly divided into sham operation (sham) group,CCH model (CCH) group, capsaicin group,and solvent group, 12 rats in each group. Capsaicin at 2.5 mg/kg was intraperitoneally injected twice a week for 4 weeks, starting on the 7th day after surgery. The rats in solvent group were given the same amount of solvent at the same time and under the same conditions. Morris water maze, object recognition test and open field test were conducted to analyze the cognitive related behavior performance on the 4th week after surgery. The changes of MAMs in the hippocampal CA1 region were observed under transmission electron microscope, the co-localization of the MAMs was observed by immunofluorescence double-labeling, and the expression of mitofusin 2 (Mfn2) in the hippocampal tissue was determined by Western blot.RESULTS:Four weeks after the operation, the behavior tests showed that the cognitive function of CCH rats was impaired compared with sham operation group. Compared with solvent group, spatial learning and memory in capsaicin group was improved significantly. The results of transmission electron microscope and confocal microscope showed that the distance of MAMs in the hippocampal CA1 area of CCH rats was increased compared with sham operation group, and the co-localization of the contacts was decreased (P<0.05). Compared with solvent group, the correlation between the mitochondria and ER in capsaicin group was increased (P<0.05). The protein level of Mfn2 in CCH group was significantly lower than that in sham group (P<0.05). Compared with solvent group, the protein level of Mfn2 in capsaicin group was higher (P<0.05). CONCLUSION:CCH rats showed decreased cognitive function and loosen MAMs. Capsaicin improves the cognitive behavior of CCH rats by up-regulation of MAMs.     

脑络通胶囊对大鼠局灶性脑缺血模型的影响

目的观察脑络通胶囊对大鼠大脑中动脉缺血模型的影响。方法通过线栓法建立大鼠大脑中动脉(MCA)局灶性脑缺血模型,动物经给药3次,观察脑络通胶囊对脑缺血大鼠的行为状态、脑缺血面积、脑组织形态学的作用。结果脑络通胶囊1.72、0.86、0.43g/kg剂量均能明显改善大鼠脑缺血后的行为状态,缩小缺血范围,降低缺血程度,使病变的脑组织得到明显地改善。     

针刺对大鼠局灶性脑缺血后神经功能缺损及 梗死面积的影响

〔摘要〕目的探讨针刺大椎、百会、人中穴对脑缺血大鼠神经功能缺损评分、脑梗死面积比的影响、方法参照 ZeaLonga线拴法复制大脑中动脉缺血模型(MCAO), 40只SD大鼠随机分为假手术组、模型组、针刺穴位组(即穴位 组,穴取大椎、百会、人中)、针刺对照点组(即对照点组,穴取大椎、百会、人中左侧旁开0.3 cm处),6次针刺后,观 察神经功能缺损评分及1`TC染色检测梗死面积比评价、结果与假手术组比较,模型组大鼠神经功能缺损评分、梗 死面积比明显增高,差异有统计学意义(P<0.01),而对穴位组、对照点组能不同程度地降低神经功能缺损评分,减少 梗死面积,差异有统计学意义(P<0.05 , P<0.01),且穴位组优于对照点组,差异有统计学意义(P<0.05 )〔结论针刺大 椎、百会、人中穴可通过改善神经功能的缺损,减少脑梗死面积从而对脑具有保护作用、     

中西医结合治疗缓慢型心律失常34例疗效观察

目的观察中西医结合治疗缓慢型心律失常疗效．方法治疗组采用中西医结合治疗,静脉滴注生脉注射液、西咪替丁等,对照组使用阿托品等治疗缓慢型心律失常．结果两组结果经统计学处理,痊愈率有高度差异性（P〈0．01）,总有效率有统计学意义（P〈0．01）．结论用中西医结合治疗缓慢型心律失常疗效显著．     

双侧脑室引流、冲洗结合腰穿救治高血压脑出血破入全脑室患者的临床观察

目的 观察双侧脑室冲洗、引流结合腰穿治疗高血压脑出血破入全脑室的有效性和安全性.方法 将高血压脑出血破入全脑室的83例分成双管组(n=57)和单管组(n=26),双管组采用双侧脑室引流、冲洗结合腰穿的方法治疗(57例);单管组用单管引流治疗(26例),将两组治疗结果进行统计学分析和比较.结果 双管组:良好和中残35例(61.4%)、重残11例(19.3%)、植物生存2例(3.5%)、死亡9例(15.8%),单管组分别为6例(23.1%)、6例(23.1%)、1例(3.8%)、13例(50.0%),以双管组的疗效为优(P＜0.01);且双管组的病死率明显低于单管组(P＜0.01).双管组的脑脊液廓清时间为3～6(4.2±1.3)d,明显短于单管组的6～15(8.6±2.1)d(P＜0.01).结论 双管引流比单管引流更容易保持引流通畅,脑脊液廓清迅速,并发症减少,能降低病死率,提高生存质量,是一种安全、有效治疗全脑室出血的方法.