大鼠颈动脉粥样硬化性狭窄模型的建立

目的 建立一种经济实用、稳定、可靠的颈动脉粥样硬化动物模型.方法 SD大鼠分为3组,每组20只.采用球囊损伤法造成颈动脉内膜损伤后,分别给予普通饲料喂养,高脂饲料灌胃,同时给予腹腔注射维生素D,于第1,2,3,4周取出损伤侧血管,行HE染色,计算各组大鼠颈动脉狭窄率和内、中膜面积.结果 注射维生素D组大鼠颈动脉损伤处,第1周时内皮下即有泡沫细胞出现,随着时间延长逐渐出现内皮细胞脱落,内膜下泡沫细胞逐渐增多,血管平滑肌细胞增生,排列紊乱,至第4周时形成明显的动脉粥样斑块,管腔狭窄,较其他组差异有统计学意义(P＜0.05).结论 SD大鼠经球囊损伤颈动脉内膜后,结合高脂饲料及维生素D腹腔注射,可形成明显的颈动脉粥样硬化斑块,该方法时间短、操作简单、成功率高,可作为脑缺血性疾病研究的动物模型.     

高脂饲料喂养与动脉内膜球囊损伤结合建立兔腹主动脉粥样硬化模型

背景：以往研究多采用单纯饲喂高脂饲料、正常或高脂血症动物动脉内膜损伤致动脉粥样硬化狭窄模型。 目的：拟采用喂养高脂饲料与动脉内膜球囊损伤相结合的方法建立腹主动脉粥样硬化的模型。 设计、时间及地点：对比观察实验,于2007-01/03在解放军总医院动物实验中心完成。 材料：新西兰白兔20只,随机分为单纯饲喂高脂饲料组、高脂饲料与动脉内膜损伤结合组,每组10只。高脂饲料由普通颗粒饲料+4%胆固醇+10%猪油组成+10%蛋黄粉组成。 方法：单纯饲喂高脂饲料组单纯喂养高脂饲料,高脂饲料与动脉内膜损伤结合组喂养高脂饲料4周后,行腹主动脉内膜球囊损伤术。 主要观察指标：12周后取主动脉行病理检查,计算机计算脂纹脂斑厚度,内、中膜厚度,内膜厚度比值,并进行血清总胆固醇、三酰甘油和高密度脂蛋白胆固醇、低密度脂蛋白胆固醇的测定。 结果：12周后,两组兔血脂血清总胆固醇、三酰甘油、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇均升高,高脂饲料与动脉内膜损伤结合组升高更明显(P < 0.05)。喂养高脂饲料的两组动物均出现动脉粥样硬化斑块,横段面纤维斑块切片出现钙化。与单纯饲喂高脂饲料组比较,高脂饲料与动脉内膜损伤术结合组内膜明显增厚, 形成了明显的斑块及纤维帽结构,斑块厚度与内中膜厚度比值增加明显(P < 0.01)。 结论：所诱导产生的斑块与人类斑块成分具有一定相似性,包括斑块的纤维帽、钙化、脂核等,提示采用动脉内膜损伤与高脂饮食结合方式来制作动脉粥样硬化斑块的模型是可行的、实用的。     

经股动脉入路建立球囊损伤兔颈动脉狭窄模型

目的经股动脉入路制作一种实用、可靠且稳定的颈动脉狭窄模型,用于探讨颈动脉再狭窄的防治。方法新西兰大白兔20只,随机分为对照组、实验1周组、实验2周组和实验4周组,每组5只,常规喂养。所有动物采用经右侧股动脉入路置入HyperForm球囊,实验组行右颈总动脉定比扩张,对照组行单纯右颈动脉路图显示,术后以普通饮食喂养。实验1、2、4周组实验兔分别于手术后1周、2周和4周处死,并在第4周同时处死对照组兔。右颈动脉分叉近心端1～2cm节段取材送光镜和电镜检查,分别观察血管壁增生情况并计算血管内膜和中膜厚度。结果与对照组比,实验组新生内膜厚度呈持续性显著增加(P<0.01),实验2周组与实验1周组比较及实验4周组与实验2周组相比,内膜厚度均显著增加(P<0.01)。各组中膜厚度未见明显统计学差异(P>0.05)。结论经股动脉入路用Hyper Form球囊行颈动脉定比扩张能成功地制作一种稳定、简便可靠的颈动脉狭窄动物实验模型。血管平滑肌细胞的过度增生是造成PTA术后管腔再狭窄的主要因素。     

颈动脉粥样硬化斑块CD147、MMP-9表达及阿托伐他汀干预的影响

目的 探讨兔颈动脉粥样硬化斑块中CD147、基质金属蛋白酶-9(MMP-9)的表达情况以及阿托伐他汀对其表达的影响.方法 24只新西兰大白兔按随机数字表法分为3组:正常对照组8只,普通饲料喂养16周;余16只给予球囊损伤颈总动脉+高胆固醇喂养,12周后分成模型对照组和阿托伐他汀组(每天2.5 mg/kg阿托伐他汀灌胃),继续喂养4周后给予鲁塞尔氏蛙蛇毒和组胺触发斑块破裂.对斑块进行病理分析,免疫组织化学法检测斑块内巨噬细胞及CD147、MMP-9蛋白表达,逆转录聚合酶链反应(RT-PCR)检测斑块内CD147、MMP-9mRNA表达.结果 药物触发后模型对照组存活的6只兔中有4只发生斑块破裂及血栓形成,阿托伐他汀组存活的7只兔中有1只发生斑块破裂及血栓形成,正常对照组中未见斑块破裂及血栓形成.与模型对照组比较,阿托伐他汀组斑块内巨噬细胞含量明显减少(33.62±3.82vs16.22±2.61),CD147蛋白表达的阳性染色面积比明显降低(29.54±3.03vs14.22±1.63),MMP-9蛋白表达的阳性染色面积比明显降低(21.18±4.75 vs 11.11±1.87),差异有统计学意义(P＜0.05＝.模型对照组和阿托伐他汀组斑块内CD147 mRNA表达水平分别为0.939±0.105和0.426±0.112,MMP-9 mRNA表达水平分别为0.335±0.069和0.135±0.067,比较差异有统计学意义(P＜0.05＝.Pearson相关检验显示斑块中MMP-9蛋白与CD147蛋白表达呈正相关(r=0.669,P=0.001),MMP-9 mRNA与CD147 mRNA表达呈强正相关(r=0.783,P=0.000).结论 CD147、MMP-9表达上调可能参与斑块不稳定的发生机制,阿托伐他汀可能通过下调CD147、MMP-9的表达发挥稳定斑块的作用.     

蛋白酶体抑制剂MG132对动脉粥样硬化的影响

目的观察蛋白酶体抑制剂MG132对动脉粥样硬化的影响。方法将新西兰白兔30只随机分成高脂组、MG132(1)组和MG132(2)组。3组兔普通饲料喂养1w后行颈动脉球囊损伤术,术后高脂饲料(含1%胆固醇、3%猪油和15%蛋黄)喂养;MG132(1)组在术后高脂饲料喂养的同时血管局部应用蛋白酶体抑制剂MG132;MG132(2)组在术后高脂饲料喂养4w后血管局部应用蛋白酶体抑制剂MG132;3组均喂养8w后取颈总动脉血管制成病理切片行HE染色。结果高脂组兔的右颈总动脉管壁呈动脉粥样硬化改变。MG132(1)组兔右颈动脉血管组织结构基本正常。MG132(2)组兔的右颈总动脉管腔介于二者之间。结论局部应用蛋白酶体抑制剂MG132能够抑制血管内膜增生及动脉粥样形成,同时可能具有稳定斑块的作用。     

银杏黄酮苷元干预兔颈总动脉损伤后内膜增生及血凝素样氧化低密度脂蛋白受体1的表达

背景：银杏叶提取物中的黄酮苷元具有较强的抗氧化活性,课题组前期实验证实可抑制氧化低密度脂蛋白诱导内皮细胞表达血凝素样氧化低密度脂蛋白受体1。 目的：进一步探讨银杏黄酮苷元对兔颈总动脉内皮损伤后内膜增生和血凝素样氧化低密度脂蛋白受体1表达的影响。 方法：将雄性新西兰大白兔随机分为对照组、假手术组、模型组和治疗组。对照组予普通饲料,其余各组予高脂饮食,假手术组仅作颈外动脉结扎,模型组和治疗组均球囊损伤右颈总动脉,治疗组损伤后用银杏黄酮苷元灌胃。4周后检测各组血脂水平,观察各组右颈总动脉形态,用免疫组织化学法和RT-PCR检测血凝素样氧化低密度脂蛋白受体1蛋白和mRNA表达。 结果与结论：模型组术后4周内膜增生明显,有粥样斑块形成,血凝素样氧化低密度脂蛋白受体1表达明显增加(P < 0.05)。治疗组银杏黄酮苷元灌胃4周后内膜增生较轻,内膜面积,血凝素样氧化低密度脂蛋白受体1阳性细胞数和其基因表达水平均低于模型组(P < 0.05),但血脂与模型组比较无差异。提示银杏黄酮苷元可减轻新生内膜增生及动脉粥样硬化病粥样斑块的形成,这种作用可能与抑制兔颈总动脉内皮损伤后血凝素样氧化低密度脂蛋白受体1的表达有关。     

渐变网格球扩支架治疗兔囊状动脉瘤

目的评估单纯使用新型渐变网格球扩支架治疗动脉瘤的可行性。方法取新西兰大白兔12只,分为实验组（n=6）和对照组（n=6）。采用弹性酶诱导方法,在右侧颈总动脉起始部制作动脉瘤模型,实验组载瘤动脉内植入渐变网格球扩支架,对照组不植入支架。观察3个月,复查造影并行统计学分析,并处死动物行病理检查。对实验组支架植入前后造影结果进行分析．拟合动脉瘤内时间－密度曲线。结果实验组载瘤动脉内均成功植入渐变网格球扩支架,其中1个动脉瘤植入后即刻和3个月造影未见动脉瘤显影,其余5个动脉瘤显影较淡;3个月时,DSA显示支架位置均良好,载瘤动脉通畅,无支架内狭窄;支架植人后时间－密度曲线与术前相比,对流相降低而弥散相增加。对照组1只动物造影前死于麻醉意外,其余5只观察3个月,动脉瘤大小无明显变化。病理显示：实验组1只不显影的动脉瘤血管内膜已完全覆盖支架,而另5只未愈合的动脉瘤颈部支架仍然裸露：对照组动脉瘤体厚薄不均匀。弹力层均消失。结论单纯使用渐变网格球扩支架可促进兔囊状动脉瘤愈合,且对载瘤动脉无明显影响。     

肩袖损伤模型兔持续被动活动对骨-肌腱界面早期修复的影响：MRI不同时点影像证实

背景：肩袖损伤中以冈上肌腱损伤最为多见,其修复的难点在于骨-肌腱界面的重建,目前缺乏合适的动物模型以及确切的治疗方法。 目的：建立兔肩袖损伤修复的实验动物模型,通过MRI观察持续被动活动对肩袖骨-肌腱界面损伤重建术后骨-肌腱界面早期修复的作用,为肩袖损伤修复术后康复制定一个合适的康复程序。 方法：8月龄雄性新西兰大白兔16只,构建双侧冈上肌腱急性损伤模型后重建其在肱骨大结节上的止点,随机分为自由活动组(n=4)和持续被动活动组(n=12)。持续被动活动模型与重建组运动术后第2天开始应用自制兔肩关节持续被动活动器,以2,4,10 (°)/s不同持续被动活动速度进行屈伸75°~75°运动。分别在术前和术后第1,2,4周观察各组动物一般情况及不同持续被动活动速度时兔肩袖骨-肌腱界面的MRI影像学变化。 结果与结论：大体观察可见重建2周后切口愈合良好,拆线后未见切口感染、血肿、裂开等;MRI检查结果显示：术后各组肩袖骨-肌腱界面逐渐愈合,术后第1,2,4周不同时间点与自由活动组相比, 持续被动活动组在不同屈伸角度下骨-肌腱界面修复迅速,每屈伸75°~75°运动速度为4 (°)/s时,骨-肌腱界面的修复明显好于自由活动组。证实该动物模型能够很好的满足实验的需要,兔冈上肌腱损伤重建术后第2天开始,不同速度持续被动活动屈伸运动,可促进骨-肌腱界面的修复,以4 (°)/s的持续被动活动速度最佳。     

MSCTA及DSA对模型兔颈动脉狭窄的评价

目的 探讨多层螺旋CT血管造影术(MSCTA)及数字减影血管造影术(DSA)对粥样硬化性颈动脉狭窄模型兔的诊断价值.方法 30只兔中的10只做为空白对照组,余20只兔颈动脉外膜置人改良的硅胶橡胶圈后,给予兔高胆固醇饲料喂养2周诱导颈动脉狭窄模型兔,采用股动脉插管方法行MSCTA及DSA检查,观察模型兔颈动脉狭窄程度及影像学征像,比较二者对颈动脉粥样硬化性狭窄的诊断价值.结果 MSCTA及DSA均可显示颈动脉管腔狭窄的部位、范围、程度及形态以及颈动脉的血管走向,MSCTA对颈动脉狭窄敏感程度较高,但对颈动脉狭窄的特异性诊断不如DSA.结论 MSCTA和DSA能较好的反应模型兔的颈动脉狭窄部位、形态.     

加味补阳还五汤对兔髂动脉球囊损伤血管狭窄和氧化应激的影响

背景：近年研究证明,补阳还五汤具有扩张血管、改善微循环、抗炎、抗氧化应激和保护血管内皮细胞的功能,但其具体机制尚不清楚,尤其是对经皮腔内冠状动脉成形后再狭窄形成过程有何影响,目前很少见报道。 目的：观察加味补阳还五汤对兔髂动脉球囊损伤后血管狭窄及氧化应激的影响。 方法：将新西兰兔以随机抽签法分为对照组、模型组和药物组。对照组给予普通饲料,模型组、药物组给予高脂饮食。饲养2周后,模型组、药物组行髂动脉内膜剥脱术,对照组兔行假手术对照,药物组术后饲料中添加加味补阳还五汤药颗粒 2 mL/(kg•d),对照组及模型组喂食同前。4周后光镜观察兔髂动脉内膜的损伤情况,并检测血脂水平、血清超氧化物歧化酶活性和丙二醛水平的变化。 结果与结论：对照组髂动脉内膜薄且结构完整,无动脉硬化斑块;模型组内膜增厚,管腔明显狭窄,可见明显动脉粥样硬化斑块;药物组动脉粥样硬化斑块厚度减小,管腔狭窄程度较轻。药物组兔血清总胆固醇、三酰甘油和低密度脂蛋白胆固醇、丙二醛水平明显低于模型组,而高密度脂蛋白胆固醇、血清超氧化物歧化酶水平明显高于模型组(P < 0.05)。结果说明加味补阳还五汤具有较好的防止家兔球囊扩张损伤髂动脉所致的管腔狭窄及抗实验性动脉粥样硬化作用,其机制可能与其清除氧自由基抗氧化应激、调节脂质代谢等作用有关。     

灯盏细辛注射液对动脉粥样硬化家兔炎症反应的影响

目的观察灯盏细辛注射液对动脉粥样硬化家兔炎症反应的抑制作用,探讨其抑制动脉粥样硬化的机制。方法健康SD雄性家兔随机分为对照组和模型组,模型组采用高脂饲料喂养联合耳缘静脉注射小牛血清白蛋白方法建立动脉粥样硬化模型,饲养10 w后,随机抽取6只进行病理形态学检测,明确动脉粥样斑块形成。然后把模型组随机分为高脂组(100 g/d)和灯盏细辛组[5 ml/(kg·d)腹腔注射]。实验10 w、18 w取血清,ELISA法测定血清肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)、C-反应蛋白(CRP)、单核细胞趋化蛋白-1(MCP-1)的水平,组织形态学方法分析动脉粥样硬化斑块情况。结果第10周,高脂组和灯盏细辛组血清TNF-α、IL-6、CRP、MCP-1水平明显高于对照组,动脉粥样硬化斑块明显形成;第18周,灯盏细辛组血清TNF-α、IL-6、CRP、MCP-1水平明显低于高脂组,动脉粥样斑块明显减轻。结论灯盏细辛注射液具有抑制、延缓动脉粥样硬化斑块进展的作用,可能与降低TNF-α、IL-6、CRP、MCP-1的分泌有关。     

内皮型一氧化氮合酶基因转染对血管损伤后新生内膜增生的影响*

背景：一氧化氮能够抑制血管平滑肌细胞的迁移和增殖,而一氧化氮合酶是其合成的关键酶,有关一氧化氮合酶基因体内转染对平滑肌细胞及动脉粥样硬化血管损伤后内膜增生影响少有报道。 目的：观察内皮型一氧化氮合酶 (endothelial nitric oxide synthase,eNOS)基因体内局部转染对动脉粥样硬化大鼠血管损伤后新生内膜增生的抑制作用。 方法：建立动脉粥样硬化Wistar大鼠颈动脉球囊损伤模型,建模后随机分成空白对照组、AdCMV-lacz对照组和AdCMV-eNOS组,分别将PBS,AdCMV-lacz和AdCMV-eNOS体内转染至以上3组大鼠的损伤血管壁。转染2周后培养并鉴定损伤局部平滑肌细胞,并用RT-PCR法检测各组损伤及转染后血管平滑肌细胞eNOS mRNA的表达,同时观察转染后不同时期新生内膜增生的影响。 结果与结论：AdCMV-eNOS组的颈总动脉血管平滑肌细胞可表达eNOS mRNA。3组大鼠转染后1和3个月,AdCMV-eNOS组内膜/中膜面积比值低于空白对照组和AdCMV-lacz对照组(P < 0.01)。结果显示,eNOS基因体内转染损伤后血管可以抑制血管新生内膜增生,减少再狭窄发生率。     

新西兰白兔非乙醇性脂肪性肝病模型的建立与评价★

背景：高脂饮食是导致非乙醇性脂肪性肝病发病的独立相关危险因素。 目的：建立非乙醇性脂肪性肝病兔模型。 方法：将新西兰白兔随机分为对照组和模型组，模型组给予高脂饮食，对照组给予普通动物饲料喂养。 结果与结论：兔饲养12周后，模型组肝细胞均呈现弥漫性脂肪变性，汇管区与小叶间可见炎细胞浸润、坏死及纤维化，对照组肝脏无异常，且模型组肝指数、丙氨酸氨基转移酶、天冬氨酸氨基转移酶、三酰甘油均高于对照组(P < 0.01或P < 0.05)。说明实验成功建立了非乙醇性脂肪性肝病兔模型。     

模拟人后外侧入路髓核摘除构建的椎间盘退行性变动物模型

背景：椎间盘退变的病理生理学机制至今尚未明了,所以制作一种与自然椎间盘退行性病变过程相类似的、适用于这些治疗方法的动物模型有助于对椎间盘退行性变的诱因、病理生理变化及某些治疗方法的研究。 目的：模拟人后路髓核摘除术进行后外侧穿刺抽吸髓核建立椎间盘退行性变动物模型的可行性研究及其优点。 方法：随机选取日本大耳白兔20只,根据解剖结构观察结果,为防止纤维环后外侧穿刺损伤血管,采用右后外侧入路,行L1~2和L3~4椎间盘后外侧穿刺髓核抽吸法摘除部分髓核组织,将L2~3椎间盘纳入对照组。术后2,4,8,12周分别对造模后椎间盘及对照椎间盘(L2~3)行X射线及MRI影像学观察。 结果与结论：日本大耳白兔20只均进入结果分析,术后2,4,8,12周实验组椎间盘高度指数持续下降(P < 0.05),说明造模后椎间盘间隙高度会逐渐降低,并且各时间点椎间盘高度指数明显低于对照组(P < 0.05)。与对照组比较,造模后2,4,8,12周时实验组椎间盘MRI T2WI信号逐渐下降,并且随时间延长逐渐降低,呈低信号改变。通过影像学观察发现本模型退行性变的征象与人腰椎间盘退行性变的征象一致,提示模拟人后路髓核摘除术建立后外侧纤维环穿刺髓核抽吸的椎间盘退行性变动物模型成功建立,为运用组织工程修复重建退行性变椎间盘的研究提供有效的动物模型。     

制作兔腰椎终板下缺血模型：MRI与病理学变化验证的可行性

背景：目前的椎间盘退变动物模型主要通过改变椎间盘生物力学环境、损伤椎间盘自身结构以及应用基因技术改变动物的遗传性状等诱发退变,但这些方法均为外界人为因素直接作用于椎间盘,与椎间盘退变的自然病程差别较大。 目的：评价经皮穿刺兔腰椎终板下椎体注射平阳霉素制作椎体终板下缺血模型的可行性。 方法：选取新西兰大白兔46只,每兔设L5为实验组、L4为对照组。穿刺腰椎终板下椎体,L5注射平阳霉素(2 g/L)1 mL,,L4注射生理盐水1 mL。其中4只兔子术前行腰动脉造影。术后第1,2,3,4,5周,2个月及3个月随机选取6只行MRI检查,并取病理送检。测量对比第4周实验组MRI与病理切片的缺血面积。 结果与结论：对照组MRI和病理学检查均无特异性变化,实验组MRI第1,2周变化不明显,第3周出现FST1WI低信号,T2WI及FST2WI呈稍高信号,第4周信号变化更明显;实验组病理第1,2周无特异性变化;第3,4周骨小梁排列杂乱, 骨细胞逐渐减少,椎体骨髓内血细胞减少,脂肪细胞增多融合,终板软骨细胞减少、结构紊乱,纤维环及髓核无明显变化;第5周出现椎间盘退变表现;第2,3个月终板下椎体缺血持续存在、椎间盘退变更加明显。实验组第4周MRI与病理切片缺血面积之间有显著正相关性(r=0.965,P < 0.001)。结果证实,采用经皮穿刺终板下椎体注射平阳霉素的方法可成功制作兔腰椎终板下缺血动物模型,具有创伤小、操作简便、重复性好和成功率高的特点。该模型是研究腰椎及椎间盘退变较理想的一种动物模型。     

Hyperlipidemia affects neuronal nitric oxide synthase expression in brains of focal cerebral ischemia rat model★

BACKGROUND: Hyperlipidemia, a risk factor for ischemic cerebrovascular disease, may mediate production of neuronal nitric oxide synthase (nNOS) to induce increased nitric oxide levels, resulting in brain neuronal injury. OBJECTIVE: To investigate effects of hyperlipidemia on brain nNOS expression, and to verify changes in infarct volume and pathology during reperfusion, as well as neuronal injury following ischemia/reperfusion in a rat model of focal cerebral ischemia. DESIGN, TIME AND SETTING: Complete, randomized grouping experiment was performed at the Laboratory of Physiology, Shanxi Medical University from March 2005 to March 2006. MATERIALS: A total of 144 eight-week-old, male, Wistar rats, weighing 160-180 g, were selected. A rat model of middle cerebral artery occlusion was established by suture method after 4 weeks of formulated diet. Nitric oxide kit and rabbit anti-rat nNOS kit were respectively purchased from Nanjing Jiancheng Bioengineering Institute, China and Wuhan Boster Biological Technology, Ltd., China. METHODS: The rats were equally and randomly divided into high-fat diet and a normal diet groups. Rats in the high-fat diet group were fed a high-fat diet, consisting of 10% egg yolk powder, 5% pork fat, and 0.5% pig bile salt combined with standard chow to create hyperlipidemia. Rats in the normal diet group were fed a standard rat chow. A total of 72 rats in both groups were randomly divided into 6 subgroups: sham-operated, 4-hour ischemia, 4-hour ischemia/2-hour reperfusion, 4-hour ischemia/4-hour reperfusion, 4-hour ischemia/6-hour reperfusion, and 4-hour ischemia/12-hour reperfusion, with 12 rats in each subgroup. MAIN OUTCOME MEASURES: nNOS expression was measured by immunohistochemistry, and pathomorphology changes were detected by hematoxylin-eosin staining. Infarct volume and nitric oxide levels were respectively measured using 2, 3, 5-triphenyltetrazolium chloride (TTC) and immunohistochemistry. RESULTS: In the ischemic region, pathology changes were significant in the 4-hour ischemia/4-hour, 4-hour ischemia/6-hour reperfusion, and 4-hour ischemia/12-hour reperfusion subgroups fed on a high-fat diet compared to the same groups fed on a normal diet. In each ischemia subgroup, nNOS expression in brain tissues was higher than in the sham-operated subgroups fed on either the high-fat diet or normal diet (P < 0.01). At each ischemia/reperfusion time point, rats fed on a high-fat diet expressed higher levels of nNOS compared to rats fed on the normal diet (P < 0.05). When tissue was stained with TTC, a white infarction area was detected in the ischemic hemisphere, demonstrating that the infarct volume gradually increased with prolonged reperfusion time in each ischemia subgroup. At each ischemia/reperfusion time point, the infarct volume was larger in rats fed on a high-fat diet compared to those fed on a normal diet. CONCLUSION: nNOS expression was greater in hyperlipidemia rats following ischemia/reperfusion. Cerebral ischemia/reperfusion injury is aggravated with prolonged reperfusion time. Key Words: focal cerebral ischemia; hyperlipidemia; ischemia/reperfusion injury; neuronal nitric oxide synthase     

Hyperlipidemia affects neuronal nitric oxide synthase expression in brains of focal cerebral ischemia rat model★

BACKGROUND: Hyperlipidemia, a risk factor for ischemic cerebrovascular disease, may mediate production of neuronal nitric oxide synthase (nNOS) to induce increased nitric oxide levels, resulting in brain neuronal injury. OBJECTIVE: To investigate effects of hyperlipidemia on brain nNOS expression, and to verify changes in infarct volume and pathology during reperfusion, as well as neuronal injury following ischemia/reperfusion in a rat model of focal cerebral ischemia. DESIGN, TIME AND SETTING: Complete, randomized grouping experiment was performed at the Laboratory of Physiology, Shanxi Medical University from March 2005 to March 2006. MATERIALS: A total of 144 eight-week-old, male, Wistar rats, weighing 160-180 g, were selected. A rat model of middle cerebral artery occlusion was established by suture method after 4 weeks of formulated diet. Nitric oxide kit and rabbit anti-rat nNOS kit were respectively purchased from Nanjing Jiancheng Bioengineering Institute, China and Wuhan Boster Biological Technology, Ltd., China. METHODS: The rats were equally and randomly divided into high-fat diet and a normal diet groups. Rats in the high-fat diet group were fed a high-fat diet, consisting of 10% egg yolk powder, 5% pork fat, and 0.5% pig bile salt combined with standard chow to create hyperlipidemia. Rats in the normal diet group were fed a standard rat chow. A total of 72 rats in both groups were randomly divided into 6 subgroups: sham-operated, 4-hour ischemia, 4-hour ischemia/2-hour reperfusion, 4-hour ischemia/4-hour reperfusion, 4-hour ischemia/6-hour reperfusion, and 4-hour ischemia/12-hour reperfusion, with 12 rats in each subgroup. MAIN OUTCOME MEASURES: nNOS expression was measured by immunohistochemistry, and pathomorphology changes were detected by hematoxylin-eosin staining. Infarct volume and nitric oxide levels were respectively measured using 2, 3, 5-triphenyltetrazolium chloride (TTC) and immunohistochemistry. RESULTS: In the ischemic region, pathology changes were significant in the 4-hour ischemia/4-hour, 4-hour ischemia/6-hour reperfusion, and 4-hour ischemia/12-hour reperfusion subgroups fed on a high-fat diet compared to the same groups fed on a normal diet. In each ischemia subgroup, nNOS expression in brain tissues was higher than in the sham-operated subgroups fed on either the high-fat diet or normal diet (P< 0.01). At each ischemia/reperfusion time point, rats fed on a high-fat diet expressed higher levels of nNOS compared to rats fed on the normal diet (P<0.05). When tissue was stained with TTC, a white infarction area was detected in the ischemic hemisphere, demonstrating that the infarct volume gradually increased with prolonged reperfusion time in each ischemia subgroup. At each ischemia/reperfusion time point, the infarct volume was larger in rats fed on a high-fat diet compared to those fed on a normal diet. CONCLUSION: nNOS expression was greater in hyperlipidemia rats following ischemia/reperfusion. Cerebral ischemia/reperfusion injury is aggravated with prolonged reperfusion time.     

经皮纤维环穿刺法建立兔椎间盘退行性变动物模型

背景：合适的椎间盘退变动物模型是椎间盘组织工程研究的必要条件,但目前尚缺乏公认的模型制备方法。 目的：采用C形臂辅助下经皮纤维环穿刺法制备兔椎间盘退变动物模型,并评估其可行性。 方法：选定新西兰大白兔L2/3和L3/4椎间盘作为穿刺干预椎间盘,L1/2和L5/6椎间盘作为空白对照组,采用C形臂辅助下经皮穿刺法干预椎间盘。于术后2,4,8,12周各选择2只兔麻醉后拍摄兔腰椎核磁共振影像,处死动物采集椎间盘,进行椎间盘髓核蛋白多糖测定。核磁共振检查观察椎间盘退行性改变,二甲基亚甲蓝染色分光光度法测定髓核中蛋白多糖含量变化。 结果与结论：术后4周穿刺干预椎间盘髓核区域核磁共振信号强度及髓核内蛋白多糖含量同空白对照组相比均下降(P < 0.05),其后两者呈现逐渐下降趋势。结果证实,C形臂X射线机辅助下经皮纤维环穿刺法可用于椎间盘退变动物模型的制备。     

丹红注射液对家兔颈动脉PTA术后内膜增生及bFGF、TF表达的影响

目的 探讨丹红注射液对家兔颈动脉经皮腔内血管成形(PTA)术后内膜增生和碱性成纤维细胞生长因子(bFGF)及组织因子(TF)表达的影响.方法 50只新西兰大白兔按随机数字表法分为5组:假手术组、模型组和低、中、高剂量丹红注射液治疗组,每组10只;每组再按检测时间点分为术后7 d、28 d 2个亚组,每亚组各5只.假手术组仅结扎颈外动脉,后四组采用球囊损伤颈动脉法制备家兔颈动脉PTA术后狭窄模型.各治疗组于球囊损伤术前第1天开始分别静脉注射丹红注射液1 mL/(kg·d)、2 mL/(kg·d)、4 mL/(kg·d),连续7 d或14 d.假手术组及模型组给予静脉注射生理盐水2 mL/(kg·d).术后7、28 d时分别取模型靶血管进行组织学观察,免疫组化染色检测bFGF和TF的表达,图像分析系统进行半定量分析.结果 术后7 d时模型组新生内膜面积、新生内膜面积/中层面积(I/M)与其他组比较差异无统计学意义(P>0.05);术后28 d时模型组新生内膜面积、I/M比值较假手术组明显增加,而中、高剂量丹红注射液治疗组较模型组均明显降低,差异有统计学意义(P<0.05).与模型组相比,中、高剂量丹红注射液治疗组血管壁bFGF和TF表达量均明显降低,差异有统计学意义(P<0.05),而低剂量丹红注射液治疗组差异无统计学意义(P>0.05).结论 中、高剂量组丹红注射液可抑制家兔颈动脉PTA术后内膜增生,其机制可能是通过下调动脉壁bFGF和TF的表达来实现的.

Abstract：

Objecfive To study the effect of Danhong injection(DI)on intimal hyperplasia and expressions of basic fibroblast growth factor(bFGF)and tissue factor(TF)in carotid arteries of rabbits after percutaneous transluminal angioplasty(PTA). Methods Fifty New Zealand White rabbits were randomly divided into 5 groups (n=10),namely sham-operated group,model group,and DI treatment groups(low-,medium-and high-dose DI treatment groups).The animals in the sham-operated group were only given external carotid artery ligation.The rabbit models of carotid stenosis in the later 4 groups were established by ballon-injury.Before the carotid balloon injury,the animals in the DI treatment groups were given DI at doses of 1 mL/kg·d-1,2mL/kg·d-1 and 4 mL/kg·d-1,respectively,for 7 or 14 d via intravenous injection;the animals in the sham-operated and model groups were given saline solution(2mL/kg·d-1)instead.The animals were sacrificed and their common carotid arteries were removed on the 7th and 28th d of surgery;morphology changes of the arterial wall with hematoxylin eosin staining were observed by optical microscope,and the expressions ofbFGF and TF of the vascular wall were detected through immunohistochemical techniques;with the help of an image analysis system,semiquantitative analysis was performed.Results On the 7th d of surgery,the intimal area and intima area/media area (I/M)ratio in the model group showed no significant differences as compared with those in the other 4groups(P>0.05).On the 28th d of surgery,the intimai area and I/M ratio in the model group were increased markedly as compared with those in the sham-operated group (P<0.05), and the intimal area and I/M ratio in the medium and high dose DI treatment groups were significantly reduced as compared with those in the in the model group (P<0.05). The expressions of TF and bFGF in the neointima of carotid arteries of rabbits treated with medium and high dose of DI were significantly weaker than those of the model group (P<0.05), and those of rabbits treated with low dose of DI had no obvious differences as compared with those of the model group (P>0.05). Conclusion Medium and high dose of DI can inhibit the intimal hyperplasia resulting from percutaneous transluminal angioplasty, might through the inhibition of expressions of bFGF and TF in the neointima of the arterial wall.