间歇性节段性动脉阻断对术后动脉内膜增生影响的实 …

目的：探讨间歇性节段性动脉阻断对阻断段动脉内膜增生的影响。方法：１６只健康杂各犬随机分成４组，解剖历侧颈总动脉，以微型动脉夹在距环状软骨各２．０cm的近、远端阻断颈总动脉０，１５，３０和６０min，术后４周取标本进行切片观察及计算机图像分析。结果：阻断１５min组阻断段动脉内膜未见明显增生，阻断３０和６０min组阻断段动脉见新生内膜形成，且后者内膜增生程度较前者严重，前者新生内膜中细胞成分多，而后者以细胞间     

间歇性节段性动脉阻断对术后动脉内膜增生影响的实验研究

目的：探讨间歇性节段性动脉阻断对阻断段动脉内膜增生的影响。方法：１６ 只健康杂种犬随机分成４ 组,解剖右侧颈总动脉,以微型动脉夹在距环状软骨各２．０ ｃｍ 的近、远端阻断颈总动脉０,１５,３０和６０ ｍ ｉｎ,术后４ 周取标本进行切片观察及计算机图像分析。结果：阻断１５ ｍ ｉｎ 组阻断段动脉内膜未见明显增生,阻断３０ 和６０ｍ ｉｎ 组阻断段动脉见新生内膜形成,且后者内膜增生程度较前者严重,前者新生内膜中细胞成分多,而后者以细胞间质为主。结论：术中节段性阻断动脉３０ ｍ ｉｎ 以上可致阻断段动脉内膜增生,故动脉阻断以１５ ｍ ｉｎ 左右为宜。     

球囊扩张兔颈总动脉致动脉再狭窄的动物模型建立

目的：本实验通过球囊扩张兔颈总动脉建立动物模型,模拟PTA对动脉壁的损伤．探讨PTA后早期动脉壁内膜、中膜变化的特点。方法：用球囊扩张兔颈总动脉建立动脉损伤动物模型,术后普通饲料喂养,在术后2d、7d、14d取手术侧动脉壁标本,切片做HE染色观察测量动脉壁内膜厚度及中膜厚度的变化情况,以同只兔未手术侧动脉做为对照组,通过统计学计算．了解其变化特点。结果：①球囊扩张后兔颈总动脉内膜厚度在术后2d、7d与对照侧相比无差异（P〉0．05）,术后,14天球囊扩张侧动脉内膜厚度与对照侧相比有明显差异（P〈0．05）;②球囊扩张后兔颈总动脉内膜出现增厚,而对照侧的内膜厚度无明显改变;③球囊扩张后兔颈总动脉中膜厚度与对照侧相比有明显差异,球囊扩张后兔颈总动脉中膜厚度在术后2d及出现增厚,术后7d及术后14d较术后2d增厚明显（P〈0．05）,在术后14d与术后7d相比无明显差异（P〉0.05）。结论：本实验动物模型的建立方法是可行的,不但维持了血管在球囊扩张后正常的血流动力学,而且对于被球囊扩张血管段的定位也有帮助。     

细胞间粘附分子-1表达对家兔动脉缺血后内膜增生的影响

目的 探讨细胞间粘附分子-1的表达对家兔动脉缺血后内膜增生的影响. 方法 选用健康家兔40只,每组10只,随机分成正常对照组和3个缺血组(缺血15 min组、缺血30 min组、缺血60 min组).分别解剖出右侧股动脉,以动脉夹钳夹使股动脉缺血段长2 cm,正常对照组只加解剖,不加阻断,缺血组分别钳夹阻断缺血15、30、及60 min后恢复血流.分别于饲养第7、14、28天后抽取外周静脉血以ELISA法检测家兔血清中的sICAM-1;同时于第28 d以免疫组化S-P法检测缺血段股动脉内皮细胞上粘附的ICAM-1及弹力纤维VG染色法计算缺血段动脉内膜、中膜厚度. 结果 ①正常对照组股动脉内皮细胞粘附的ICAM-1呈弱阳性表达(2.80±0.35),与缺血15 min组相比无显著性差异(P＜0.05),缺血15 min与缺血30 min(26.67±0.75)相比差异显著(P＜0.01),缺血30 min组明显低于缺血60 min组(35.38±0.65)(P＜0.01).②正常对照组、缺血15 min组股动脉内膜无增生;缺血30 min组见动脉内膜明显增生,与缺血60 min组相比差异显著(P＜0.01). 结论 动脉缺血后通过测定ICAM-1值判定内膜增生程度与ICAM-1值有关.     

低温预防周围神经缺血再灌注损伤的形态学研究

采用暂时阻断家兔髂总动脉起始处的血流的实验模型,观察用降温方法预防坐有神经的缺血再灌注损伤（缺血４小时,再灌注５小时）的形态学变化,结果显示：神经的缺血再灌注损伤在低温下较常温下明显轻微,提示降温可预防周围神经缺血再灌注损伤。     

兔缺血再灌注脊髓损伤节段病理变化的定位研究

目的 观察并比较两种动物模型中脊髓各节段病理变化。方法 采用腰动脉阻断法和腹主动脉阻断法造成兔脊髓缺血再灌注损伤，然后对损伤脊髓各节段标记定位，进行病理学观察。结果 腰动脉阻断法中以L3、4节段损伤员重，其近侧及远侧节段损伤逐渐减轻；腹主动脉法中从L2至脊髓末端均有严重损伤；T4节段变有轻度病理改变。结论 腰动脉阻断法中脊髓末端仍存在血供，损伤范围小；腹主动脉法中则没有血供，损伤范围大；严重的脊髓缺血再灌注损伤可波及到非缺血区组织。     

自体静脉Cuff减轻动脉内膜增生的实验研究

目的：探讨聚四氟乙烯（ＰＴＦＥ）血管加自体静脉Ｃuff吻合对动脉内膜增生的缓解作用 及对流出道的改善作用。方法：健康杂种犬１２只（２４条腿），随机分成对照组产验组。对照组动脉阻断后以ＰＴＦＥ血管直接行旁路转流术；实验组以自体颈外静脉Ｃrff间置于ＰＴＦＥ与远端股动脉之间。术后第８周行动脉造影，制备标本行组织学观察及计算机图像分析，测定增生内膜、中膜的厚度、增生内膜的截面积、血管腔截面积及两者的比值、静脉     

家兔不同脑区神经元对全脑缺血再灌注损伤的敏感性

目的；研究家兔不同脑区神经元对全脑缺血再灌注损伤的敏感性。方法；结扎家兔左侧椎动脉，夹闭双侧颈总动脉４０ｍｉｎ，松开再灌注４８ｈ，７２ｈ。光镜观察。结果：神经元呈缺血性改变。海马，纹状体损伤严重，皮层病变轻微。结论；不同脑区神经元对缺血再灌流损伤的敏感性与其动脉分布有关。     

洋金花总生物碱对缺血再灌注脑组织病理形态和丙二醛的影响

目的在阻断双侧颈总动脉和椎动脉结合低血压法复制的脑完全性缺血再灌注模型上，研究了缺血再灌注脑组织病理形态和丙二醛含量变化的关系。结果提示：洋金花总生物碱能抑制膜脂质过氧化作用，使缺血再灌注脑组织的丙二醛含量和病理形态改变减轻。动态观察血液丙二醛，可作为评价脑缺血性疾病治疗和预后的监测指标。     

阿魏酸钠对球囊损伤大鼠颈总动脉血管内膜增生的影响

目的：研究阿魏酸钠对球囊损伤大鼠颈总动脉内膜增生的影响。方法：制作球囊损伤的大鼠颈总动脉内膜增生模型，Wistar雄性大鼠15只随机分为①对照组(n=7)腹腔内注射生理盐水(4ml／d)。②阿魏酸钠治疗组(n=8)腹腔内注射融于生理盐水的阿魏酸钠(200mg/d)，球囊损伤14d后，损伤区域的血管段取材固定后，分析血管断面组织形态学的变化；免疫组化方法观察增殖细胞核抗原(PCNA)阳性细胞以了解血管壁细胞的增殖情况。结果：阿魏酸钠可明显减少新生内膜面积，增加管腔面积，抑制平滑肌细胞的增殖。结论：阿魏酸钠具有抑制血管内膜增生的作用。     

大鼠新型颈总动脉球囊损伤后狭窄模型的建立及其动态病理学观察

目的为经皮腔内冠状动脉成形术后再狭窄的研究提供新型的造模方法。方法SD大鼠24只，假手术组6只，模型组18只（分3个时相点5d、14d和28d，每个时相点6只动物）。用2．0Fogarty球囊导管自右股动脉插管建立大鼠左颈总动脉球囊损伤模型。每组动物于相应时相点麻醉心脏放血处死后，取损伤颈总动脉。常规固定、包埋、切片，分别做HE染色、TR-A02弹力纤维染色和Masson三色染色，在光学显微镜下病理观察。结果球囊损伤大鼠颈总动脉后，5d可见内皮细胞脱落，表面炎性细胞浸润，附壁少量血栓形成。随着时间的延长，14d动脉内膜逐渐增厚，VSMC增殖。胶原和弹力纤维组织大量增生，28d可见内膜明显不规则增生，管腔明显变窄。结论本实验成功的复制了球囊损伤术后血管狭窄动物模型，其病理改变与临床经皮腔内冠状动脉成形术后再狭窄的病理改变相似。而且该造模方法与自颈外动脉插管建立颈总动脉再狭窄模型相比失血少。创面小，操作简便迅速，有利于提高手术成功率。     

腺病毒介导的AT2R基因转染对培养大鼠颈动脉内膜增生的影响

目的构建AT2R重组腺病毒载体并研究AT2R对动脉损伤后新生内膜增生的影响.方法将AT2R基因克隆至腺病毒载体,构建AT2R重组腺病毒;球囊损伤大鼠颈动脉后立即取下颈动脉,用重组腺病毒转染,在体外培养2周后,观察AT2R基因转染对损伤动脉新生内膜形成的影响.结果构建的AT2R重组腺病毒经鉴定正确,其滴度为1×109pfu/ml;AT2R重组腺病毒转染可明显抑制培养大鼠颈动脉新生内膜增生.结论成功构建了AT2R重组腺病毒载体;AT2R可明显抑制大鼠颈动脉球囊损伤后的新生内膜增生.     

Experimental Study of Adenovirus Vector Mediated-hVEGF165 Gene on Prevention of Restenosis after Angioplasty

This study evaluated the effects of adenovirus vector mediated human vascular endotheli-al growth factor-165 (hVEGF165) gene on prevention of restenosis after angioplasty. Rabbit models of bilateral carotid artery injury were established by balloon denudation. The recombinant adenovi-ruses containing hVEGF165 cDNA was directly injected into left side of the injured carotid arteries.On day 3 and week 3 after transfection the expression of VEGF was observed by RT-PCR and im-munohistochemistry. The thrombokinesis, reendothelialization (rET) and intimal hyperplasia in ca-rotid arteries were evaluated by computerized image analysis system 3 weeks after gene transfer,The changes in the VEGF gene-treated side were compared with the control side. Our results showed that 3 days and 3 weeks after hVEGF165 gene transfer the VEGF mRNA and antigen ex-pression were detected in vivo. 3 weeks after the transfer, the carotid artery rET was markedly better in the VEGF gene-treated group compared with the control. The thrombokinesis, intima are-a/media area (I/M), maximal intimal and medial thicknesses (ITmax and MTmax) demonstrated a statistically significant decrease in arteries treated with VEGF gene as compared with the control group. It is concluded that VEGF gene transfer could be achieved by intra-arterial injection of re-combinant adenoviruses. It might accelerate the restoration of endothelial integrity, inhibit throm-bokinesis and attenuate intimal hyperplasia in the injured arteries after VEGF gene transfer. This procedure could be useful in preventing restenosis after angioplasty.     

激活剂蛋白-1诱骗性寡核苷酸对大鼠颈总动脉损伤后新生内膜的影响

目的探讨激活剂蛋白1(AP1)诱骗性寡核苷酸对大鼠颈总动脉损伤后新生内膜的影响及其作用机制。方法36只Wistar大鼠随机分为3组,每组12只,均建立颈总动脉损伤模型,单纯损伤组不予任何处理;Lipofectin转染试剂组局部释放Lipofectin;治疗组局部释放AP1诱骗性寡核苷酸,3组均于于术后30min、3h、7d处死13取材,观察比较内膜增生程度,免疫组化方法检测增殖细胞核抗原(PCNA)和c fos。结果单纯损伤组和Lipofectin转染试剂组7d时可见内膜明显增生,且有较多平滑肌细胞表达PCNA,c fos30min表达达高峰。而治疗组内膜增生程度减轻,PCNA、c fos的表达率明显下降,差异有显著意义(P<0.05或P<0.01)。结论局部转染AP1诱骗性寡核苷酸可明显抑制动脉损伤后的内膜增生,下调PCNA、c fos的表达。     

缬沙坦对大鼠颈动脉球囊损伤后血管内膜增生和TLR4表达的影响

目的:观察缬沙坦对大鼠颈动脉球囊损伤后血管内膜增生及Toll样受体4(toll like receptor4,TLR4)表达的影响。方法:以1.5F球囊导管建立大鼠颈动脉球囊损伤模型,SD大鼠随机分为假手术组(n=6)、球囊损伤组(n=12)、缬沙坦组(n=12)。造模后第1天开始给予缬沙坦10mg.kg-1.d-1灌胃,假手术组及球囊损伤组灌胃等量蒸馏水,14d后取损伤血管段,组织形态学观察并测定内膜增生情况,实时定量PCR检测血管TLR4mRNA的表达。结果:术后14d,球囊损伤组和缬沙坦组可见明显内膜增生,内膜面积及内膜和中膜面积比均显著大于假手术组,而缬沙坦组上述指标均低于球囊损伤组(P<0.05);球囊损伤组和缬沙坦组TLR4mRNA的表达较假手术组升高,与球囊损伤组比较,缬沙坦可降低球囊损伤诱导的TLR4高表达。结论:缬沙坦可抑制大鼠颈动脉球囊损伤引起的血管内膜增生,其作用可能是通过下调损伤血管TLR4的过表达而实现。     

Experimental study of adenovirus vector mediated-h VEGF<Subscript>165</Subscript> gene on prevention of restenosis after angioplasty

Summary This study evaluated the effects of adenovirus vector mediated human vascular endothelial growth factor-165 (hVEGF₁₆₅) gene on prevention of restenosis after angioplasty. Rabbit models of bilateral carotid artery injury were established by balloon denudation. The recombinant adenoviruses containing hVEGF₁₆₅ cDNA was directly injected into left side of the injured carotid arteries. On day 3 and week 3 after transfection the expression of VEGF was observed by RT-PCR and immunohistochemistry. The thrombokinesis, reendothelialization (rET) and intimal hyperplasia in carotid arteries were evaluated by computerized image analysis system 3 weeks after gene transfer. The changes in the VEGF gene-treated side were compared with the control side. Our results showed that 3 days and 3 weeks after hVEGF₁₆₅ gene transfer the VEGF mRNA and antigen expression were detectedin vivo. 3 weeks after the transfer, the carotid artery rET was markedly better in the VEGF gene-treated group compared with the control. The thrombokinesis, intima area/media area (I/M), maximal intimal and medial thicknesses (IT_max and MT_max) demonstrated a statistically significant decrease in arteries treated with VEGF gene as compared with the control group. It is concluded that VEGF gene transfer could be achieved by intra-arterial injection of recombinant adenoviruses. It might accelerate the restoration of endothelial integrity, inhibit thrombokinesis and attenuate intimal hyperplasia in the injured arteries after VEGF gene transfer. This procedure could be useful in preventing restenosis after angioplasty. Liu Qigong, male, born in 1968, Associate Professor This project was supported by a grant from Wuhan Chenguang Program (No. 20015005048)     

瑞舒伐他汀对损伤动脉一氧化氮合成酶的影响

目的观察瑞舒伐他汀对大鼠颈总动脉球囊损伤后内膜增生及内皮化的影响,探讨瑞舒伐他汀对内皮型一氧化氮合酶（eNOS）及诱生型一氧化氮合酶（iNOS）的影响,评价瑞舒伐他汀对再狭窄的影响与一氧化氮合成酶的关系。方法 48只大鼠随机分为3组,每组24只,即瑞舒伐他汀组：于术前3d开始连续每天予瑞舒伐他汀5mg/kg？d灌胃;损伤组：予等量生理盐水灌胃;对照组（对照组与损伤组共用大鼠,损伤组取材左侧损伤颈总动脉,对照组取右侧正常颈总动脉）。3组各于术后即刻、7d、14d、28d麻醉并处死大鼠,留取两侧颈总动脉标本,应用组织形态学方法检测内膜增生并进行计算机图像分析,RT-PCR方法检测eNOS及iNOS的表达情况。结果瑞舒伐他汀组内膜面积低于损伤组;损伤组eNOSmRNA术后明显低于对照组,iNOSmRNA表达高于对照组;瑞舒伐他汀组eNOSmRNA表达明显高于损伤组,iNOSmRNA表达明显低于损伤组（p〈0.05）。结论瑞舒伐他汀可抑制内膜增生,对球囊损伤具有修复作用,这可能与瑞舒伐他汀调节一氧化氮合成酶的表达,促进大鼠颈总动脉球囊损伤后再内皮化有关。     

大鼠颈动脉损伤后内膜增生的计算机图像分析

目的　利用计算机图像处理分析系统对局部应用反义寡脱氧核苷酸( ODNs) 对大鼠颈动脉损伤后内膜增生抑制作用进行定量分析。方法　将大鼠随机分成6 组( 即反义组A、正义组B、错配组C、对照组D、脂质体组E、假手术组F) ,进行大鼠颈动脉损伤后内膜增生的形态参数测定。结果　A 组管腔面积大于B、C、D 及E 组( P< 0 .05) 且小于F 组( P< 0 .05) ;A 组的内膜面积、内膜/ 中膜面积比值、最大内膜厚度、内膜/ 中膜厚度比值、内膜覆盖率均明显小于B、C、D 及E组( P< 0 .01) ,而B、C、D 及E 组组间两两比较无显著性差异( P> 0 .05) 。结论　利用计算机图像处理从量化角度分析内膜增生的各种情况,进一步证实了血管紧张素Ⅱ1 型受体( AT1R) 的反义( ODNs) 对氮气干燥后大鼠颈动脉内膜增殖有抑制作用,且具有序列依赖性     

Inhibitory effect of adenoviral vector-mediated AT2R gene transfection on neointimal hyperplasia

Objective： To investigate the effect of adenoviral vector-mediated AT2R gene transfection on neointimal hyperplasia after rat carotid artery balloon injury. Methods ：AT2R gene was transferred into rat carotid arteries by recombinant adenovirus pAd-AT2R after the establishment of rat carotid balloon injury restenosis model. The arteries were harvested on the 14th day after gene transfer. The efficiency of transgene delivery was measured by the expression of adenovirus-encoding green fluorescent protein （GFP） under fluorescent microscope. The expression of AT2R and PCNA （proliferating cell nuclear antigen） was evaluated by RT-PCR, immunocytochemistry, immunofluorescence staining, confocal microscopy, respectively. The ratio of intimal to medial area （I/M） was quantified with images and determined by an image analysis system. Results, GFP-positive area in adventitia, media and the forming neointima was about 40%. Adenoviral delivery of rat AT2R gene up-regulated AT2R expression in balloon-injured rat carotid and reduced PCNA expression and I/M significantly in neointima（P〈0.01）. Double immunofluorescence labeling of AT2R and PCNA also showed that AT2R gene transfer inhibited VSMCs proliferation in neointima. Conclusion： AT2R gene transfer may be a novel promising therapy to limit neointimal hyperplasia.