链脲佐菌素所致高血糖对大鼠缺血性脑损伤的影响

目的 研究链脲佐菌素所致糖尿病性高血糖（ＨＧ）对大鼠缺血性脑损伤的影响。方法 链脲佐菌素引起糖尿病４周的大鼠造成１０ｍｉｎ前脑缺血,观察缺血后脑组织的病理改变和癫痫发作情况。结果 在海马ＣＡ１区和下脚区,ＨＧ组与正常血糖（ＮＧ）组动物脑神经元损伤程度相似。然而,ＨＧ组动物有严重的顶叶皮质神经元坏死和“额外脑结构损伤”。ＨＧ组中４２．１％的动物脑缺血后出现癫痫发作,血浆葡萄糖浓度低于１２ｍｍｏｌ／Ｌ     

链脲佐菌素侧脑室注射后大鼠海马SYP改变

目的研究侧脑室内注射链脲佐菌素（streptozotocin,STZ）对大鼠海马突触素（synaptophysin,SYP）的影响。方法将40只SD大鼠随机分成对照组和模型组,于第1d和第3d侧脑室注射STZ3rag／kg建立大鼠AD模型,15d后进行Morris水迷宫试验,取海马后通过RT-PCR、western-blotting及免疫组织化学法检测SYP表达水平。结果与对照组相比较,侧脑室注射STZ后大鼠逃避潜伏期明显延长,穿越平台次数明显减少,SYP表达明显减少。结论侧脑室注射STZ后大鼠空间记忆力减退与突触结构受损有关。     

侧脑室注射链脲佐菌素对大鼠脑内能量代谢及线粒体功能影响的研究

目的探讨侧脑室注射链脲佐菌素(streptozotocin,STZ)对大鼠脑内能量代谢及线粒体功能的影响。方法 12只SD大鼠随机分为STZ组及生理盐水组。30天后电镜下观察两组大鼠海马及额顶颞皮层,并检测线粒体呼吸链复合物Ⅱ和Ⅳ、Na+ -K+ -ATP酶、活性,及三磷酸腺苷(adenosine triphosphate,ATP)、二磷酸腺苷(adeno- sine diphosphate,ADP)的浓度。结果 STZ组海马及额顶颢皮层电镜下有明显改变,ATP浓度、ATP/ADP比值及线粒体功能降低明显(P>0．05),ADP浓度升高明显(P>0．05)。结论侧脑室注射STZ引起能量代谢明显异常及线粒体活性的下降并有神经细胞受损。     

链脲佐菌素剂量对C57BL/6J小鼠糖尿病神经病理性疼痛诱导效应的影响

目的观察不同剂量链脲佐菌素(STZ)对C57BL/6J小鼠糖尿病神经病理性痛诱导效应的影响,探讨其量效关系及最佳剂量。方法雄性C57BL/6J小鼠85只,随机分为正常对照组(n=10)和A～E 5个实验组(n=15,STZ腹腔注射剂量分别为80、100、120、140、160mg/kg),对照组腹腔注射等体积柠檬酸缓冲液。观察各组血糖、机械痛阈值、热刺痛阈值和28d生存率的变化,分析其与STZ剂量的关系。结果 A～E各组糖尿病成模率均>86.7%。C组神经病理性疼痛成模率(66.7%)均高于其余各组;E组28d生存率(46.7%)与A、B组间均存在显著差异(P<0.05)。结论 C57BL/6J小鼠腹腔注射STZ以120mg/kg为最佳剂量。     

链脲佐菌素对人神经母细胞瘤系SH—SYSY细胞生长的影响

目的 研究体外链脲佐菌素(STZ)对人神经母细胞瘤(SH-SY5Y)细胞生长以及SH-SY5Y细胞胰岛素信号转导通路相关蛋白表达的影响.方法 采用MTT测定法测定SH-SY5Y细胞活性,乳酸脱氢酶(LDH)漏出率测定法观察SH-SY5Y细胞生长情况;应用Western blot法检测胰岛素信号转导通路相关蛋白胰岛素受体-1(IRS-1)、磷脂酰肌醇激酶-3(PI3K)等的改变.结果 STZ与SH-SY5Y细胞共同孵育可抑制SH-SY5Y细胞生长,阻断胰岛素对SH-SY5Y细胞的促生长作用,且STZ抑制细胞生长呈明显的时间-剂量效应.随STZ浓度增加,LDH漏出率也增加.Western blot半定量分析发现IRS-1、PI3K表达减少.结论 体外STZ与SH-SY5Y细胞共孵育可能影响胰岛素/胰岛素样生长因子-1信号转导系统对细胞的促生长作用.STZ与SH-SY5Y细胞共孵育可能作为体外胰岛素信号转导通路的一种细胞模型应用于某些神经药理学研究.     

链脲佐菌素对大鼠海马p-CREB表达的影响及APP5肽类似物165的治疗作用

目的探讨链脲佐菌素(streptozotocin,STZ)对大鼠海马磷酸化环磷腺苷反应成分结合蛋白(phosphatedcylinc AMP response element binding protein,p-CREB)表达的影响以及APP5肽类似物165的治疗作用。方法将42只Wistar大鼠随机分为对照组、模型组和APP5肽类似物165治疗组。将STZ溶解于人工脑脊液,新鲜配制成浓度为25mg/mL,分别于第1、3天按体重3mg/kg行双侧脑室注射。APP5肽治疗组于3周后开始行APP5肽类似物165灌胃,连续4周。对照组和模型组则以等量生理盐水灌胃。4周后取脑组织海马行免疫组织化学染色和Western-blotting检测p-CREB。结果模型组p-CREB阳性神经细胞数(137.44±22.62)较对照组(27.33±9.91)明显增多(P<0.05),胞质淡染;APP5肽治疗组p-CREB阳性神经元数目(29.78±10.72)与对照组比较差异无统计学意义(P>0.05)。模型组海马p-CREB表达多于对照组和APP5肽治疗组,APP5肽治疗组与对照组接近。结论脑室注射小剂量STZ可使海马p-CREB阳性神经元表达增多,而APP5肽类似物165可使p-CREB阳性神经元表达恢复至正常水平。     

上皮膜蛋白1在实验性糖尿病大鼠模型中额叶皮质血—脑屏障上的表达

目的测定不同时间点实验性糖尿病大鼠额叶皮质血—脑屏障(BBB)上皮膜蛋白1(EMP1)的表达。方法选成年健康雄性Sprague-Dawley大鼠随机分为链脲佐菌素(STZ)诱导糖尿病组(STZ组)和对照组(CON组),每组分为2w、4w、8w 3个亚组,采用免疫组织化学法检测EMP1的表达。结果STZ大鼠与CON大鼠比较,4w组中STZ大鼠上EMP1的表达显著增多(P0.01),2w组和8w组中差异无统计学意义;EMP1的表达在STZ大鼠各亚组之间及CON大鼠各亚组之间比较差异无统计学意义(P0.05)。结论 EMP1在实验性糖尿病大鼠额叶皮质BBB上的表达改变,在模型建立成功后4w时表达显著增加。     

糖尿病大鼠周围神经形态、功能及电生理指标的动态变化

目的观察链尿佐菌素(STZ)诱导的1型糖尿病大鼠周围神经病变的发生、发展变化。方法用STZ(60mg/kg)腹腔注射诱导1型糖尿病大鼠模型,选取造模成功的大鼠随机分为造模后2周、4周、6周及8周组,对照组给予腹腔注射生理盐水,亦随机分为2周、4周、6周及8周组,每组8只。测定大鼠各时间点热痛阈值、尾神经感觉神经传导速度及腓肠神经的超微结构,观察以上3个指标的动态变化。结果 STZ诱导糖尿病大鼠模型制作成功率为83.3%。造模后4周,模型组热缩足反应潜伏期(TWL)明显长于对照组(P0.05),尾神经感觉传导速度明显慢于对照组(P0.05),模型组少数腓肠神经有髓纤维髓鞘扭曲,部分髓鞘板层松散。造模后6周,模型组TWL显著长于对照组(P0.01),尾神经传导速度显著慢于对照组(P0.01),模型组多数腓肠神经有髓纤维髓鞘板层松散明显,轴突变性。造模后8周,模型组腓肠神经有髓纤维可见典型髓球样改变。结论糖尿病大鼠周围神经功能改变早于形态改变。STZ造模后6周,周围神经形态和功能均有显著改变。     

股骨头骨骺缺血性坏死模型兔构建及组织病理学变化和p53与bcl-2的表达

背景：研究表明儿童股骨头骨骺缺血性坏死(Perthes病)的发病与儿童股骨头血液供应、髋关节周围病变、外伤、内分泌因素、遗传等多种原因有关。 目的：建立可靠的Perthes病的动物模型,并观察其股骨头组织学变化、细胞凋亡情况及p53和bcl-2基因的表达,并探讨凋亡基因p53和bcl-2与Perthes病的的相关性。 方法：采用C型臂透视下经右侧股骨颈注射TH胶致骺板缺血,建立幼兔Perthes病模型,并以同样方法用经右侧股骨颈给予注射生理盐水的兔作对照,于建模后4,6,8,10,12周分别取模型组和对照组两组右侧股骨头做病理切片,观察两组髋关节骺板细胞凋亡情况,并观察两组右侧股骨头p53和bcl-2等凋亡基因表达阳性率的变化。 结果与结论：建模后4周开始出现模型组右侧髋关节关节间隙增宽,出现点状出血变性区并逐步扩大,随后关节软骨与骨骺的黏附强度下降,骨质松脆,干骺端疏松变软。模型组右侧股骨头骺板细胞凋亡率与p53基因表达阳性率呈明显正相关性(r =0.68, P < 0.05),与bcl-2基因表达阳性率之间呈明显负相关性(r =-0.75, P < 0.01)。实验建立的perthes病动物模型稳定,简单,无反复现象。结果证实了Perthes病的发生与p53和bcl-2凋亡基因相关。     

心肌缺血再灌注损伤模型大鼠心肌组织及血清肌钙蛋白Ⅰ表达和心肌肿瘤坏死因子α与重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白的干预**

背景：心肌缺血再灌注时生成大量的肿瘤坏死因子α直接造成心肌的收缩功能下降。 目的：观察药物重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白对大鼠缺血再灌注心肌损伤的影响。 方法：成年雄性Wistar大鼠建立心肌缺血再灌注模型。药物干预组在再灌注前注射重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白,模型组给予生理盐水,并设立不造模的假手术组。再灌注后立即测量心肌梗死面积,ELISA检测再灌注后的心肌肿瘤坏死因子α及血清肌钙蛋白Ⅰ的含量,实时PCR检测心肌肿瘤坏死因子α mRNA的表达。 结果与结论：与假手术组相比,模型组与药物干预组大鼠心肌肿瘤坏死因子α及其mRNA和肌钙蛋白的水平明显升高 (P < 0.05);与模型组相比,药物干预组心肌肿瘤坏死因子α及血清肌钙蛋白Ⅰ水平明显升高(P < 0. 05),心肌梗死体积与肿瘤坏死因子α mRNA的表达减少(P < 0. 05)。提示重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白能够减轻心肌缺血/再灌注损伤作用,改善大鼠的心功能。     

构建股骨头缺血坏死模型兔总胆固醇和三酰甘油的变化

背景：引起股骨头坏死最常见的非创伤性致病因素是激素的大量使用,尽管激素与非创伤性股骨头坏死的关系密切,但其发病机制至今尚未阐明。 目的：通过建立股骨头缺血坏死动物模型并检测脂质代谢指标,以探讨脂质代谢与股骨头坏死的关系。 方法：20只健康日本大白兔随机分成2组。实验组通过联合应用马血清和甲泼尼龙建立股骨头坏死的动物模型。对照组腹腔注射生理盐水。于开始注射甲泼尼龙针后第2,4,6,8周检测血清中总胆固醇和三酰甘油;于第2,4,6周分别处死实验组及对照组各2只动物行苏木精-伊红染色组织学检查;于第8周,剩余动物行DR、CT检查后处死行组织学检查。 结果与结论：实验组DR检查显示双侧骨密度降低,骨骺线不清,骨质网状结构模糊;CT显示骨质网状结构密度降低,呈点状分布,骨皮质变薄,骨质疏松,骨骺线不清;苏木精-伊红染色显示骨小梁结构不清,排列紊乱,骨髓内脂肪细胞增多,骨细胞缺失,血栓形成,肥大细胞增生,而对照组的DR、CT和苏木精-伊红染色未见明显异常。实验组造模后总胆固醇、三酰甘油升高(P < 0.05),且明显高于对照组(P < 0.01)。结果证实,联合应用马血清和激素可成功建立股骨头坏死动物模型。脂质代谢紊乱是激素性股骨头坏死重要发病机制之一。     

链脲酶素诱导灵长类动物的糖尿病模型

背景：为了提高人类同种胰岛移植的临床疗效，于临床前研究设计良好大型动物模型如猪或灵长类动物是至关重要的。 目的：评价不同剂量链脲酶素诱导非人类灵长类1型糖尿病模型的效能。 设计、时间及地点：对比观察动物实验，于2007-10/2008-12在中南大学湘雅三医院细胞移植与基因治疗中心完成。 材料：成年健康雄性恒河猴21只分为：125 mg/kg链脲酶素组(n=5)、75 mg/kg链脲酶素组(n=5)、50 mg/kg链脲酶素组(n=11)。 方法：根据动物体质量，称取一定量链脲酶素，用预先配制好的缓冲液，配制为25 g/L，过滤消毒。将新鲜配置的链脲酶素按125，75，50 mg/kg剂量分别注入3组实验猴静脉内，药物注射时间为1~5 min。 主要观察指标：观察用药后1~16周动物的肝、肾功能，葡萄糖代谢和组织形态学变化。 结果：在125 mg/kg剂量下，由于胰腺β细胞严重受损， 2只恒河猴在给予链脲菌素后8 h因严重低血糖死亡；该组恒河猴给药1周出现肝脏转氨酶及血肌酐、尿素氮的显著升高，2~4周达峰值，1只恒河猴于链脲酶素给药后由于内源性胰岛素严重不足，外源性胰岛素不能纠正其高血糖状态，最终发生严重的糖代谢紊乱、酮症酸中毒、肝肾功能衰竭于第13天死亡，存活2只恒河猴在观察期间内肝脏转氨酶及血尿素氮、血肌酐均维持在较高水平；在75 mg/kg剂量下，恒河猴在链脲酶素给药一两周出现肝脏转氨酶及血肌酐、尿素氮的显著升高，4周后肝肾功能仍表现为不同程度的异常，其中1只恒河猴于注入链脲酶素后出现肾功能损害及机体抵抗力低下，出现眼睑肿胀、全身浮肿及臀部感染不愈，于链脲酶素给药后5周而死亡，1只动物于注入链脲酶素后出现持续高血糖状态不能纠正、食欲不振，体质量显著下降，因全身衰竭而于注药后第9周死亡；在给予50 mg/kg的剂量，肝肾功能的影响较小，表现为一过性轻度升高，4周后基本可恢复正常范围，该组仅3只动物注药后1~4周出现眼睑水肿，随后水肿消退。 结论：低剂量50 mg/kg 链脲酶素可能是构建多数恒河猴糖尿病模型的最佳剂量。     

激素性兔股骨头坏死模型的建立

背景：激素的应用已经成为激素性股骨头缺血坏死发病的首要原因。 目的：拟应用马血清与皮质醇激素联合制备兔股骨头缺血性坏死早期模型,探讨激素性股骨头坏死的发病机制。 方法：新西兰大白兔随机分成3组。激素联合马血清组静脉注射马血清10 mL/kg,3周后再次注射马血清6 mL/kg,再2周后注射甲强龙45 mg/kg,1次/d,连续5 d。激素组注射甲强龙45 mg/kg,1次/d,连续5 d。正常对照组不做任何处理。于激素给予前及激素注射后1,3,7和14 d检测定血胆固醇、三酰甘油水平;于激素注射后第2、4、8周行股骨头MRI和组织病理学检测。 结果与结论：与对照组比较,激素联合马血清组和激素组大白兔血清三酰甘油和总胆固醇分别于激素注射后1 d 和3 d明显高于对照组(P < 0.01)。MRI检测结果显示,激素联合马血清组大白兔股骨头于激素注射后第4周出现坏死信号;激素组第8周出现坏死信号。组织学检测结果显示,激素联合马血清组大白兔于激素注射第4周时股骨头出现骨小梁部分变细、断裂,空骨陷窝增加;第8周骨小梁稀疏、破碎,脂肪细胞增大,空骨陷窝明显增大。激素组病理坏死程度各时段均较激素联合马血清组轻。结果表明,激素联合马血清方法可成功制备股骨头坏死早期模型。     

酒精性和激素性股骨头坏死的发病与脂质代谢

背景：非创伤性股骨头坏死的发病机制存在各种理论学说,脂质代谢异常是其中一种假说,但是脂质代谢如何参与骨坏死的发病机制仍不清。 目的：通过对激素性和酒精性骨股头坏死患者血脂改变及股骨头置换标本形态计量学观察分析,探讨脂质代谢改变在非创伤性骨坏死中的作用。 方法：选择股骨头坏死住院患者162例,包括酒精性骨股头坏死57例,激素性骨股头坏死63例,创伤性骨股头坏死42例,其国际骨循环分期为(ARCO)Ⅱ~Ⅳ期。患者入院后常规抽空腹静脉血行三酰甘油、低密度脂蛋白等血脂检测。各组选取12例关节置换股骨头标本,股骨头冠状面正中剖开,在负重区和非负重区取材,对骨髓造血面积、脂肪细胞密度、脂肪细胞直径和脂肪细胞面积等行形态计量学观察。 结果与结论：酒精性股骨头坏死患者血三酰甘油升高率显著高于激素性和创伤性股骨头坏死患者(P < 0.05)。酒精性股骨头坏死患者血低密度脂蛋白降低率明显高于激素性和创伤性股骨头坏死(P < 0.05)。与激素性骨坏死患者相比,酒精性股骨头坏死患者脂肪细胞面积和脂肪细胞直径大,但造血面积减小,且差异有显著性意义。推测脂质代谢改变在酒精性股骨头坏死晚期起重要调节作用,而在激素性股骨头坏死晚期作用不明显。     

Changes of NADPH-diaphorase reactivity in lumbar spinal cord of short-term streptozotocin induced diabetic rats

Diabetes is an endocrine and metabolic disorder often associated with erectile dysfunction and peripheral neuropathy. Among other factors, penile erection is induced by activation of nitric oxide synthase (NOS). Hypothalamic paraventricular nuclei neurons produce NO and project to spinal cord areas implicated in penile reflexes. These nuclei have shown an increase of NOS in streptozotocin-induced diabetic rats. NOS-containing neurons are identical to the populations of neurons selectively stained for NADPH-diaphorase activity. Using this technique, we have evaluated changes of NOS in the lumbar spinal cord of diabetic rats with or without insulin treatment. Positive staining was found in motoneurons, dorsal horn neurons (layer II), neurons surrounding the ependimus (layer X) and neurons at the intermediolateral cell column (ILCC). Diabetic animals showed significant decrease in reactive area and increase of the histochemical reaction in motoneurons from the sexual dimorphic nuclei and in neurons of the ILCC. A marked decrease of the number of reactive neurons was also observed in layer II. Morphologic alterations were observed in neurons of layer X as an increase in the percentage of multipolar neurons and a decrease in the number and length of secondary processes. The alterations observed in these animals were absent in the insulin treated diabetic animals. These results show the plasticity of lumbar spinal cord neurons, suggesting a direct participation of NO synthesis in the physiopathology of the erection dysfunction in diabetes.     

成人非创伤性股骨头坏死股骨头塌陷的预测

背景：股骨头坏死塌陷就意味着软骨下骨板的力学性能失效，最终会导致受累的髋关节功能障碍。 目的：测量真实坏死股骨头的坏死体积及坏死体积占整个股骨头体积的百分比，分析股骨头坏死病灶大小和部位与股骨头塌陷的关系。 设计、时间及地点：单一样本观察，于2006-10/2007-10在河北工程大学附属医院解剖教研室完成。 对象：选择河北工程大学附属医院骨科收治的股骨头坏死患者12例15髋，男9例，女3例，年龄41~59岁。 方法：将坏死股骨头用1 mm厚手动钢锯冠状位均匀劈成10等份，将锯开的股骨头放入标有刻度的量杯中，加入生理盐水测量股骨头体积；在每层断面上仔细辨认股骨头坏死反应性增生的硬化带，其内测缘即为股骨头坏死的边界，用尖嘴咬骨钳咬除股骨头坏死部分，然后将去除坏死后的股骨头放入量杯中，加入生理盐水测量股骨头坏死体积。 主要观察指标：①股骨头坏死体积及坏死体积比测量结果。②坏死体积比例与坏死股骨头数目关系。 结果：股骨头劈开后测量整个股骨头的平均体积为50.0 cm3，股骨头坏死区域的平均体积为21.6 cm3，股骨头坏死区域的体积与整个股骨头的体积比平均值为43.5%。股骨头坏死体积比在40%以上的有9髋，30%~40%有4髋，30%以下的有2髋。 结论：坏死病灶的大小与股骨头塌陷的危险性密切相关，当坏死病灶体积大于30%时，股骨头坏死的塌陷率高达87%，具有高风险塌陷；小于30%时，坏死病灶的位置对病情进展起关键作用。     

Changes in nucleotide hydrolysis in rat blood serum induced by pentylenetetrazol-kindling

There is growing pharmacological evidence from several animal models of seizure disorders that adenosine possesses endogenous anticonvulsant activity. Apart from being released from cells, adenosine can be produced by the degradation of adenine nucleotides by ectoenzymes or soluble nucleotidases. These enzymes constitute an important mechanism in synaptic modulation, as they hydrolyze ATP, an excitatory neurotransmitter, to adenosine, a neuroprotective compound. We recently demonstrated an increase in ectoenzyme activity in rat brain synaptosomes after pentylenetetrazol-kindling in rats resistant to kindling, suggesting a role for ectonucleotidases in the seizure control. The present work investigates the effect of seizures induced by pentylenetetrazol kindling on the enzymes that could be playing a role in ATP, ADP and AMP hydrolysis to adenosine in rat blood serum. Animals received injections of PTZ (30 mg/kg, i.p., dissolved in 0.9% saline) once every 48 h, totaling 10 stimulations and the controls animals were injected with saline. The hydrolysis of ATP, ADP and AMP were significantly increased (42, 40, and 45%, respectively), while phosphodiesterase activity was unchanged. These results suggest once more that an increase in the ATP diphosphohydrolase and 5'-nucleotidase activities and, possibly, in adenosine levels, could represent an important compensatory mechanism in the development of chronic epilepsy. Moreover, the fact that this increase can also be measured in serum could mean that these enzymes might be useful as plasma markers of seizures in epilepsy.     

构建酒精性股骨头坏死动物模型的理论依据及造模方法*

背景：酒精性股骨头坏死晚期股骨头易塌陷、致残率高,且目前缺乏有效的防治方法,故备受国内外学者的关注,因此,建立酒精性股骨头坏死动物模型,是进一步探讨其发病机制及临床防治的重要研究方向。 目的：总结国内外对酒精性股骨头坏死动物模型建立的研究进展。 方法：应用计算机分别在PubMed数据库和中国期刊全文数据库(CNKI)中检索1990-01/2010-01有关酒精性股骨头坏死的临床和基础实验研究的文章,检索词分别为“alcohol intoxication; avascular necrosis; animal model; research progress”和“酒精中毒;股骨头坏死;动物模型;研究进展”,纳入所述内容与酒精性股骨头坏死的动物模型相关的文献,排除发表时间较早或重复性研究。 结果与结论：收集到151篇相关文献,排除121篇不符合标准的文献,共纳入30篇符合标准的文献。经分析得出以下结论：通过对鼠、兔等动物过度饮酒或股骨头局部酒精介入,可导致股骨头髓内脂肪细胞增多、增大,小血管数量减少或阻塞,造成微循环障碍导致其缺血、缺氧而最终发生坏死,其发生机制接近于人类股骨头缺血性坏死的早期病理改变。结合国内外对酒精性股骨头坏死动物模型的研究现状,采用灌胃法,给予中国白兔烈性中国白酒(含体积分数55%乙醇)8 mL/(kg•d)4~8周处理,此种造模方法更适合目前实验研究。     

Changes in regional energy metabolism after closed head injury in the rat

We examined in the present investigation regional ATP, glucose, and lactate content in the cortical and subcortical structures, in a rat model of closed head injury (CHI). In serial tissue sections bioluminescence imaging of ATP, glucose, and lactate was performed at 4 h, 12 h and 24 h (n=4/5 per time point with) after the induction of CHI or sham surgery. Bioluminescence images were analyzed by computer-assisted densitometry, at the lesion site, in remote cortical areas, and in the subcortical structures (thalamus and caudate nucleus). ATP content was significantly decreased at the lesion site after 4 h and in the remote cortex at 12 h post-injury. At 12 h, the ATP content reached baseline levels on the ipsilateral side and at 24 h also at remote lateral parietal sites. In the contralateral cortex, ATP increased transiently above the baseline at 12 h. No significant changes in ATP were found in the thalamus and caudate nucleus. Cortical glucose and lactate contents could not be discerned over time. Following CHI there is an acute and progressive, yet transient, ischemic cortical profile, which is not reflected in subcortical areas.     

Changes in tumor necrosis factor alpha and myeloper-oxidase in mouse models of local cerebral infarction induced by photochemical method

BACKGROUND: Lots of evidences have demonstrated that acute inflammatory reaction plays an important role in cerebral ischemia and cerebral ischemia/reperfusion injury. Tumor necrosis factor (TNF), as one of important inflammatory cytokines, also participates in the injury. OBJECTIVE: To observe the changes in TNF-α expression and myeloperoxidase (MPO) activity of mouse models of local cerebral infarction induced by photochemical method, and analyze the correlation of TNF-α expression and MPO activity. DESIGN: Randomized controlled experiment. SETTING: Laboratory of Cerebral Microcirculation, Taishan Medical College. MATERIALS: Sixty involved male adult Kunming mice were provided by the Experimental Animal Center of Shandong University. TNF-α primary antibody, kits for enzyme-linked immunosorbent assay(ELISA) and streptavidin-biotin complex immunohistochemical dyeing kit were purchased from Boster Company(Wuhan). MPO kit was purchased from Jiancheng Bioengineering Institute (Nanjing). Cold light source was developed by Hengfa Co.,Ltd.( LG-150, Xuzhou). METHODS: This experiment was carried out in the Laboratory of Cerebral Microcirculation of Taishan Medical College between July 2004 and July 2005. The involved 60 Kunming mice were randomized into 3 groups: normal control group (n =6), sham-operation group (n =6) and model group (n =48). Mice in the model group were observed at 30 minutes, 1, 3, 6, 12, 24, 48 and 72 hours after illumination, separately, 6 mice at each time point. In the model group, mice models of local cerebral infarction were developed as follows: The mice were anesthetized to expose left skulls. Taking 2 mm left to sagittal suture and 2 mm posterior to coronal suture as center, a field with diameter of 3 mm for illumination was set. The optical fiber detecting head of cold light source was vertically close to exposed skull. The mice were injected with rose Bengal for 5 minutes, and then cold light source was open for 10 minutes. Illumination was omitted in the sham-operation group. Mice in the control group were not modeled. At postoperative 6 hours, TNF-α expression in infracted-side cortex was detected with immunohistochemical method and ELISA, and MPO activity in infracted-side cortex with chromatometry. MPO activity could reflect the infiltration degree of neutrophils in tissue. Stronger activity indicated severer infiltration. Single-factor analysis of variance was used for comparison among groups, q test for pairwise comparison and correlative analysis for detecting the inter-parameter correlation. MAIN OUTCOME MEASURES: Changes in TNF-α expression and MPO activity of left cortex of mice in each group. RESULTS: Sixty mice were involved in the final analysis. After cerebral infarction, TNF-α positive cells were neurons and glial cells mainly, distributing in and around the infarct region. TNF-α expression in cortex of mice of sham-operation group was （615.7±16.1） ng/L, and that of model group increased to （792.2±17.8） ng/L at 3 hours after illumination, and reached peak [（921.9±23.9） ng/L] at 6 hours after illumination, and decreased to （848.0±30.6） ng/L at 12 hours after illumination and recovered to the normal level [（625.3±14.3） ng/L] at 72 hours after illumination. MPO activity of sham-operation group was （7.151±0.433） nkat/g, and that of model group increased to （10.469±0.600） nkat/g at 3 hours after illumination, reached the peak [（15.486±0.650） nkat/g] at 12 hours after illumination, decreased to （11.052±0.617） nkat/g at 24 hours after illumination and recovered to the normal level [（7.418±0.617） nkat/g] at 72 hours after illumination. Change of MPO activity lagged behind that of TNF-α, and correlative analysis showed that the both were positively correlated（r =0.953, P < 0.01）. CONCLUSION: In the acute stage of cerebral infarction of mice induced by photochemical method, TNF-α expression in infarcted-side cortex is closely related with infiltration of neutrophils. TNF-α induces inflammatory cells to intrude into ischemic brain tissue, and participates in the inflammatory reaction process at the early stage of cerebral ischemia.