Therapeutic angiogenesis induced by human hepatocyte growth factor (HGF) gene in rat myocardial ischemia models

Coronary arteriosclerotic cardiopathy is also named myocardial ischemia, which severely threatens humanhealth. Following the economic development and change of life style in China, population blood pressure, weight index and serum cholesterol level all rise. This prophesies incidence rate of coronary arteriosclerotic cardiopathy and stroke will increase year by year. Angioplasty and surgical bypass, the primary interventional therapies for these in-dividuals, are temporally limited by the prob…     

Early association of electrocardiogram alteration with infarct size and cardiac function after myocardial infarction

OBJECTIVE: Myocardial infarction (MI) is the main cause of heart failure, but the relationship between the extent of MI and cardiac function has not been clearly determined. The present study was undertaken to investigate early changes in the electrocardiogram associated with infarct size and cardiac function after MI. METHODS: MI was induced by ligating the left anterior descending coronary artery in rats. Electrocardiograms, echocardiographs and hemodynamic parameters were assessed and myocardial infarct size was measured from mid-transverse sections stained with Masson's trichrome. RESULTS: The sum of pathological Q wave amplitudes was strongly correlated with myocardial infarct size (r = 0.920, P < 0.0001), left ventricular ejection fraction (r = -0.868, P < 0.0001) and left ventricular end diastolic pressure (r = 0.835, P < 0.0004). Furthermore, there was close relationship between MI size and cardiac function as assessed by left ventricular ejection fraction (r = -0.913, P < 0.0001) and left ventricular end diastolic pressure (r = 0.893, P < 0.0001). CONCLUSION: The sum of pathological Q wave amplitudes after MI can be used to estimate the extent of MI as well as cardiac function.     

大鼠心肌缺血模型在环境毒理学中应用的探讨

结扎大鼠左冠状动脉的左室支,造成局部心肌缺血,大体标本活性体染色与光镜检查的结果相符合,均显示缺血中心区大部分心肌梗塞,变性;缺血损伤在导致心电图紊乱的同时,还使心肌组织的脂质过氧化产物丙二醛大量生成和自由基清除剂超氧化物歧化酶的活性显著下降;     

磁场对缺氧性心肌损伤大鼠心肌MDA含量变化的影响

目的：探讨磁场对缺氧性心肌损伤大鼠心肌MDA含量变化的影响。方法：本实验将大鼠随机分为四组：空白对照组、磁作用对照组、缺氧性心肌损伤组、缺氧性心肌损伤磁场治疗组。采用硫代巴比妥酸法测定各组大鼠心肌MDA含量。结果：心肌MDA含量,缺氧性心肌损伤磁场治疗组明显低于缺氧性心肌损伤组（P＜0．01）。结论：磁场对缺氧性心肌损伤大鼠心肌MDA含量具有明显降低的作用。初步证实,磁砀对缺氧性心肌损伤大鼠心肌具有一定的保护作用。进而表明,磁场对克山病心肌具有一定的保护作用。     

极化液对大鼠不同血糖浓度下心肌缺血再灌注的影响

目的:探讨不同血糖浓度下大鼠心肌缺血再灌注(MI/R)心肌损伤的程度及明确极化液(GIK)对高血糖心肌MI/R的保护作用.方法:将12只健康SD大鼠,随机分为4组,每组3只,即对照组、GIK组、高血糖组、GIK+高血糖组.每组大鼠结扎左冠状动脉前降支30 min,再灌注120 min.采用氯化三苯基四氮唑(TTC)染色法检测大鼠缺血再灌注心肌的心肌梗死面积.动态观察Ⅱ导联心电图QRS波、T波、ST段和心率变化.结果:TTC染色显示每组大鼠心肌都有梗死,心肌梗死面积高血糖组>GIK+高血糖组>对照组>GIK组;心电图显示高血糖大鼠心肌缺血期ST段明显抬高,再灌注后实验动物出现心律失常.GIK对正常血糖大鼠MI/R具有明显保护作用.结论:GIK对高血糖大鼠心肌缺血再灌注损伤保护作用有限,而对正常血糖大鼠MI/R损伤有较好的保护作用.     

磁场对缺氧性心肌损伤大鼠血红细胞SOD活性变化的影响

目的：探讨磁场对缺氧性心肌病大鼠血红细胞SOD活性变化的影响。方法：将大鼠随机分为四组：空白对照组、磁作用对照组、缺氧性心肌损伤组、缺氧性心肌损伤磁场治疗组。采用四氮唑蓝法测定各组大鼠血红细胞SOD活性。结果：大鼠血红细胞SOD活性,心肌损伤磁场治疗组明显高于缺氧性心肌损伤组（P＜0．01）,磁作用对照组高于空白对照组（P＜0．05）。结论：在磁场作用下缺氧性心肌损伤大鼠和健康大鼠血红细胞SOD活性可明显升高,进而表明：磁场对缺氧性心肌损伤大鼠和健康大鼠心肌具有一定的保护作用。     

黄芪丹参配伍提取物对心肌缺血大鼠的心脏保护作用

为证实黄芪丹参配伍提取物对心肌缺血模型大鼠的心脏具有保护作用,通过左前降支结扎诱导心肌梗死（MI）大鼠模型的方法,研究培哚普利组（PB组）、黄芪组（HQ组）、丹参组（DS组）、黄芪丹参配伍提取物组（HQ+DS组）等药物干预后对心功能的影响。采用Notch信号抑制剂RO4929097探讨Notch信号在HQ+DS组诱导心肌梗死边缘区（infarcted border zone,IBZ）血管生成中的作用。用动物超声和Masson染色评价左室射血分数（left ventricular ejection fraction,LVEF）和心肌梗死面积百分比。用免疫荧光法测定心肌IBZ中CD31和vWF的平均光密度（average optical densities,AODs）。Western blot法检测低氧诱导因子1-??（hypoxia inducible factor 1-alpha,HIF-1??）、成纤维细胞生长因子受体1（fibroblast growth factor receptor 1,FGFR-1）、Notch1和Notch胞内结构域（Notch intracellular domain,NICD）等血管生成相关蛋白和干细胞动员相关蛋白,例如基质细胞衍生因子1（stromal cell-derived factor 1,SDF-1）、C-X-C趋化因子受体4型（C-X-C chemokine receptor type 4,CXCR-4）和心肌营养素1（cardiotrophin 1,CT-1）。结果表明：与模型组相比,治疗3周后HQ+DS和PB组的LVEF均明显改善、心肌梗死面积降低,尽管与模型组相比HQ组和DS组LVEF增加、心肌梗死面积减少,但差异不显著;模型组和HQ+DS-I组IBZ中CD31的AODs均明显低于假手术组,与模型组相比,HQ+DS显著增加IBZ中CD31的 AODs,降低梗死区CD31和vWF的AODs;模型组和各治疗组HIF-1的表达均高于假手术组;与模型组相比,HQ+DS组的FGFR-1、SDF-1、CT-1、Notch1和NICD表达增加;与HQ+DS组相比,HQ+DS-I组的Notch1和NICD表达降低。可见黄芪丹参配伍对IBZ心肌细胞的保护作用优于黄芪或丹参单独应用,黄芪丹参配伍保护MI大鼠的的机制可能通过Notch信号传导和动员干细胞向IBZ移动而发挥促血管新生作用。     

磁场对缺氧性心肌损伤大鼠血浆MDA含量变化的影响

目的：探讨磁场对缺氧性心肌损伤大鼠血浆MDA含量变化的影响。方法：将大鼠随机分为四组：空白对照组、磁作用对照组、缺氧性心肌损伤组、缺氧性心肌损伤磁场治疗组。采用硫代巴比妥酸法测定各组大鼠血浆MDA含量。结果：血浆MDA含量,缺氧性心肌损伤磁场治疗组明显低于缺氧性心肌损伤组（P＜0．01）。结论：磁场对缺氧性心肌损伤大鼠血浆和MDA含量具有明显降低的作用。初步证实,磁场对缺氧性心肌损伤大鼠及克山病心肌具有一定的保护作用。     

心肌缺血后处理对急性心肌梗死患者血清CK和CK-MB的影响

目的探讨缺血后处理对急性心肌梗死(AMI)患者血清CK和CK-MB峰值及其动态变化的影响.方法将64例12 h内行直接经皮冠状动脉介入治疗(PCI)的ST段抬高型AMI患者随机分为两组,对照组34例,缺血后处理组30例.对照组给予单纯再灌注治疗,缺血后处理组采用再灌注30 s/再缺血30 s,交替3次后再持续灌注的方法...     

Effects of Chinese herbs capable of replenishing qi, nourishing yin and activating blood circulation and their compatibility on differentially expressed genes of ischemic myocardium

This study was conducted to investigate the effects of Chinese herbs capable of replenishing qi, nourishing yin and activating blood circulation and their compatibility on differentially expressed genes of ischemic myocardium which were selected from differential expression profile we had established before, and to explore the underlying mechanism. The acute myocardial infarction （AMi） model was established by ligating the left anterior descending （LAD） coronary artery, then the model rats were randomly divided into the model group, the Metoprolol group, the replenishing qi nourishing yin （RN） group, the activating blood circulation （AB） group, and the replenishing qi, nourishing yin and activating blood circulation （RA） group. In addition, the normal group and the sham group were set up. The rats of medication groups were administered by intragastric gavage with corresponding drugs on the second day after operations, and the rats of the normal group and the sham group were given normal saline as the same time.Then the ischemic hearts were harvested on the 8th day after operation. The myocardial pathomorphological changes were observed under a light microscope. The mRNA changes of target genes such as COX5a and ATP5e were detected using Real-time fluorescence quantitative PCR （Q-PCR）, and the activities of related enzymes were detected by colorimetric assay. The main resuits were as follows： the histological changes were observed by HE staining, and cardiocyte swelling, inflammatory cell infiltration and cytolysis were showed in regional ischemic myocardium of the model group, while the pathomorphological changes in all medication groups did not show obvious changes. Two genes related to energy metabolism, COX5a and ATP5e, were selected as the target genes which were down-regulated at the mRNA level in the medication groups. The activities of correlative functional enzymes also decreased in the RA group compared to that in the model group accordingly （P〈0.05）. The results indicated that the abnormal expression of genes involved in energy metabolism pathways could be one of the molecular mechanisms of AMI. The compatibility of Chinese herbs capable of replenishing qi, nourishing yin and activating blood circulation affects the expression of energy-relative gene COX5a, ATP5e, which is probably the mechanism of myocardial preservation, and is more effective than single herb of replenishing qi and nourishing yin or activating blood circulation.     

Detecting myocardial ischemia with 2-D CVIB imaging method--an in vivo animal experiment study

A 2-D cyclic variation of integrated backscatter (CVIB) imaging method was established for detecting myocardial ischemia. To demonstrate the feasibility and validity of this method, animal experiments were conducted. Acute myocardial ischemia was induced by occluding left anterior descending coronary artery in 10 anesthetized open-chest dogs. While scanning the normal hearts and the ischemic hearts with a B scanner, digital radiofrequency data were acquired by a real-time acquisition system in synchronism. The offline analysis to the radio-frequency signal with the 2-D CVIB imaging method was performed to verify the consistency between the imaging result and the design of the experiment. In addition, 4 dogs in experiment were treated with the heart pacemaker in order to investigate the influence of changing in heart rate on the detection of ischemic myocardium with the proposed method. The experimental result showed that the 2-D CVIB imaging method succeeded in detecting the ischemic myocardium and is a new non-invasive way for the cardiologists to both quantitatively and visually evaluate the contractile performance of the myocardium.     

重组腺病毒Canstatin对人肝癌HepG2细胞增殖影响的研究

为研究重组腺病毒Canstatin感染人肝癌HepG2细胞及细胞转染后Canstatin在HepG2细胞中的表达,探讨Canstatin基因对人肝癌HepG2细胞生长增殖的影响。采用不同感染复数(MOI=10、40、80)的腺病毒Ad-Canstatin-GFP感染HepG2细胞,倒置荧光显微镜下观察细胞形态,流式检测感染效率,Real-Time PCR和Western blot法检测HepG2细胞中Canstatin mRNA和蛋白表达。CCK-8法检测细胞增殖能力的变化,Western blot法检测细胞中PCNA蛋白的表达。结果显示MOI为40时,72 h感染率可达到98.9%,HepG2细胞中Canstatin mRNA和蛋白表达均高于对照组和空载体组(P0.05)。HepG2细胞感染Ad-Canstatin腺病毒后生长增殖受到抑制(P0.05),且PCNA的表达低于对照组(P0.05)。由此可知,Canstatin抑制人肝癌HepG2细胞的生长增殖有作用可能与影响PCNA的表达有关。     

云南省野生恒河猴三种病毒血清抗体的初步调查

用酶免疫法（ＥＩＡ）检查了云南省３６０只野生恒河猴的轮状病毒、腺病毒和痘病毒血清抗体,抗体阳性率分别为８６７％,８０８％和１９４％．成年猴上述三种病毒抗体的阳性率明显高于未成年猴．在云南省思茅、文山和临沧地区三种病毒抗体阳性率无明显差异,其中轮状病毒、腺病毒抗体阳性率较高,痘病毒抗体阳性率较低．结果表明：轮状病毒、腺病毒和痘病毒抗体阳性动物在野外广泛存在,抗体阳性动物的分布与年龄有关．在云南省思茅、文山和临沧三个地区,三种病毒的抗体阳性率无差异．     

CAV-1感染MDCK中各IFN及受体分子表达谱的差异

目的 分析犬肾细胞系(MDCK)感染犬腺病毒1型(CAV-1)后各种干扰素(IFN)亚型及其受体分子转录水平的表达情况。方法 建立犬 IFN-β、IFN-β 受体(IFNAR1)、IFN-λ1、IFN-λ1 受体(IFN-λ1R)、IFN-λ3 和 IFN-λ3 受 体(IFN-λ3R)的染料法定量PCR方法,分析了 MDCK在Poly I：C处理和接种CAV-1后细胞内各IFN及受体分子转录水平的相对含量。结果 正常MDCK细胞内存在较高含量的IFN-X3R且含量显著高于其他IFN及受体分子的 转录水平;用Poly I：C处理MDCK细胞24 h后,IFN-X1R、IFN-X3和IFN-X3R的表达量均显著性上调,而IFN-λ1R、IFN-λ3和IFN-λ3R几乎没变化;接种CAV-1后,IFN-α的含量急剧上升至6倍。结论 MDCK细胞存在不同水平浓度的 IFN分子及其受体转录产物,且在Poly I：C刺激和腺病毒感染时受体分子的表达谱不同。     

CTGF mRNA在糖尿病大鼠心肌中的表达及贝那普利的干预

目的研究结缔组织生长因子(CTGF)mRNA在糖尿病(DM)大鼠心肌中的表达及贝那普利(Benazepril)的干预作用.方法wistar大鼠34只建立糖尿病模型后,随机分为贝那普利治疗组及糖尿病未治疗组,12周末应用Rt-PCR检测糖尿病大鼠心肌CTGR mRNA的表达,并与正常组对照.结果DM未治疗组CTGR mRNA的表达明显高于正常组和贝那普利治疗组(P＜0.01).结论持续高糖能导致心肌CTGF mRNA表达增强,而贝那普利能在一定程度上抑制其表达,从而可能减轻糖尿病心肌病的病理变化.     

高原环境心肌细胞体外生长特性研究

目的：研究高原环境心肌细胞体外生长特性,作为高原环境研究心肌细胞功能的实验基础。方法：将心肌细胞系复苏后使用体积分数为0.15的高糖DMEM培养基接种于33mm培养皿中并置于37℃5%CO2培养箱中进行培养;台盼蓝染色法检测细胞的成活率,并观察心肌细胞的形态变化,CCK8法绘制细胞生长曲线,血球计数法进行细胞计数。结果：心肌细胞最初接种时呈圆形,24小时后细胞贴壁生长;4天后细胞有伪足伸出,细胞呈梭形、多角形等形态,大小均匀,生长迅速,7天后细胞逐渐形成细胞簇,11天后细胞簇相互连接成片生长,达90%融合时进行传代;计数1000个细胞,蓝染细胞48个,成活率为95.20%;生长曲线显示细胞生长过程分为潜伏期、对数生长期和平台期三个阶段。结论：高原环境能够进行体外心肌细胞培养。     

Therapeutic angiogenesis induced by human hepatocyte growth factor gene in hindlimb ischemia of dogs

A preclinical study of treating peripheral srtery occlusive disease(PAD) was performed by using a hepatocyte growth factor(HGF) gene-expressing vector,plasmid pUDKH,in a dog model with complete ischemia of one hindlimb.After ligation of femoral artery of one hindlimb,pUDKH was transferred directly into the ischemic limb muscles.The angiogenic activity of the plasmid pUDKH was evaluated.On D 30 after injecting once of pUDKH at differ-ent doses into local muscles immediately after operation,the degree of augmentation of collateral vessel formation was significantly greater than that treated by blank vector.In addition,the blood flow rate of femoral artery in dogs treated with pUDKH was recovered on D90,while the folw rate was only 1/5 tp 1/3 in control dogs.The pulse amplitude of pUDKH-treated dogs was recovered on D90,but it was hardly detectable in most of the control dogs.The side effects of intramuscular transfection of pUDKH were also investi-gated,and no significant positive change was found.It is suggested that angiogenesis induced by HGF gene has the potential for clincal use in the treatment of peripheral arte-rial diseases.     

西红花甙对缺血性脑梗塞的药效研究

西红花甙经十二指肠给药，对电凝阻断大脑脑中动脉造成的局灶性脑缺血有明显保护作用，能明显缩小脑梗塞灶，使梗塞后动物的活动行为障碍减轻，同时脑指数、ＭＤＡ含量明显降低。     

大鼠急性心肌梗塞后在不同部位移植胚胎干细胞疗效研究

目的探讨大鼠急性心肌梗塞（AMI）后在不同部位移植胚胎干细胞（ESC）的心肌组织形态学及血液动力学变化。方法Wistar大鼠40只随机分为正常对照组、梗塞未治疗组（梗塞组）、梗塞中心移植组（中心组）、梗塞周边移植组（周边组）共4组。结扎冠状动脉左前降支制成急性心肌梗塞模型,梗塞后1周移植体外分化并经标记的ESCs,移植后4周分别检测组织形态及血流动力学指标的改变。结果周边组移植细胞稳定存活,而中心组移植细胞未能存活。心功能及组织学检测表明中心组与梗塞组无显著差异（P〉0．05）;与梗塞组比较,周边组梗塞面积显著小于梗塞组（P〈0．01）,（21．0±1．3）％VS（40．7±2．2）％;左室重量小于梗塞组（P〈0．01）,（702．0±24．0）nagVS（882．2±32．6）mg;反应左室收缩功能的指标左室内压最大上升速率和左室内压均大于梗塞组（P〈0．01）,分别为（7．9±0．7）X103mmHg／sVS（5．9±0．5）×103mmHg／s和（117．5±10．7）mmHgVS（89．24-8．1）mmHg;而左室舒张末期压力均明显小于梗塞组（P〈0．01）,（8．5±0．3）mmHgV8（13．6±1．2）mmI-Ig。结论梗塞周边区移植ESCs可以阻止心室重构、减少瘢痕面积、改善心功能。     

电针治疗对脑梗死大鼠nNOS免疫阳性神经元表达的影响

目的 :研究电针对脑梗死大鼠尾壳核、海马CA1区缺血半影 (IP)区神经元型一氧化氮合酶 (nNOS)免疫阳性神经元的影响 ,为探讨针刺治疗脑梗死神经机制提供实验依据。方法 :采用线栓法建立大鼠永久性大脑中动脉栓塞的脑梗死模型 ,分为缺血组和缺血 +电针组 ;用免疫组化的方法观察尾壳核、海马CA1区IP区nNOS免疫阳性神经元表达的变化。结果 :(1)缺血组 :缺血 1d ,尾壳核IP区nNOS免疫阳性神经元深染固缩 ;缺血 7d后 ,阳性神经元数目增加 (P <0 0 5 ) ;在海马CA1区IP区 ,缺血 1d ,阳性神经元分布广 ;缺血 7d ,数目减少 (P <0 0 5 ) ;缺血 14d ,数目较缺血7d时增加 (P <0 0 5 )。 (2 )缺血 +电针组 :尾壳核IP区nNOS免疫阳性神经元数目较缺血组增加(P <0 0 5 ) ;在海马CA1区IP区 ,电针 1d ,阳性神经元分布局限 ,数目较缺血组减少 (P <0 0 1) ,电针 7d后 ,数目较缺血组增多 (P <0 0 5 )。结论 :电针对nNOS免疫阳性神经元调节具有双重性。既能减少电针 1d时海马IP区nNOS表达 ,减轻一氧化氮 (NO)的毒性作用 ;又能加速海马 (电针 7、14d)、尾壳核IP区nNOS表达的恢复