培养的大鼠血管平滑肌细胞中bFGF基因表达与血管紧张素Ⅱ …

采用Ｎｏｒｔｈｅｒｎ杂交检测培养的自发性高血压大鼠（ＳＨＲ）和正常血压ＷＫＹ大鼠的血管平滑肌细胞（ＶＳＭＣ）中碱性成纤维细胞生长因子（ｂＦＧＦ）基因表达，用紫外法和放免法分别测定培养液中血管紧张素转换酶（ＡＣＥ）活性和血管紧张素Ⅱ（ＡｎｇⅡ）含量，发现ＡｎｇⅡ能明显促进ＶＳＭＣ中ｂＦＧＦ基因表达，而ｂＦＧＦ则能明显诱导ＡＣＥ活性和提高ＡｎｇⅡ释放，且ＳＨＲＶＳＭＣ的ｂＦＧＦ基因表达，ＡＣＥ活性和Ａ     

几种生长因子及血管内皮细胞生长因子受体在胃肠道平滑肌肉瘤中的表达

本研究是应用免疫组化法观察细胞因子血管内皮细胞生长因子（ＶＥＧＦ）、血小板衍生生长因子（ＰＤＧＦｂ）、转化生长因子（ＴＧＦβ）、碱性成纤维细胞生长因子（ＢＦＧＦ）及血管内皮细胞生长因子受体（ＶＥＧＦＲ）在平滑肌肉瘤组织和ＥＬＩＳＡ法在细胞株的表...     

自发性高血压大鼠左心室肥厚与局部胰岛素样生长因子Ⅰ的关系

目的和方法：采用放射免疫分析法测定自发性高血压大鼠（ＳＨＲ）和正常血压大鼠（ＷＫＹ）心肌组织中胰同素样生长因子Ⅰ（ＩＧＦ－Ｉ）水平,用心肌细胞横径（ＴＤＭ）和心脏湿重／体重（ＨＷ／ＢＷ）判定左心室肥厚。结果：ＳＨＲ的ＴＤＭ和ＨＷ／ＢＷ均十分显著的高于ＷＫＹ;心肌组织ＩＧＦ－Ｉ水平也显著高于ＷＫＹ,且ＳＨＲ心肌组织ＩＧＦ－１与ＴＤＭ及ＨＷ／ＢＷ之间均呈显著正相关。结论：ＳＨＲ有明显的左心室肥厚,而组     

VEGF-B和VEGF-C的研究进展

ＶＥＧＦＢ 和ＶＥＧＦＣ 是近来发现的血管内皮生长因子（ＶＥＧＦ） 家族的两个新成员,ＶＥＧＦＢ有两种存在形式,即ＶＥＧＦＢ１６７ 和ＶＥＧＦＢ１６８ ,主要表达于胚胎及成人的肌肉组织（ 心肌及平滑肌） ,可通过与ＶＥＧＦＲ１／ＦＬＴ１ 结合发挥促血管生成的作用。ＶＥＧＦＣ 主要表达于胚胎中富含淋巴管的区域及成人的心脏、胎盘、卵巢、小肠和甲状腺等,能通过与ＶＥＧＦＲ２ 及ＶＥＧＦＲ３／ＦＬＴ４ 结合,发挥促血管、淋巴管形成及维持淋巴内皮细胞功能的作用。二者的发现使ＶＥＧＦ家族的成员进一步扩大并说明了内皮细胞功能调节的复杂性。     

血管内皮生长因子及其受体在卵巢癌中表达的免疫组化 …

目的;检测血管内皮生长因子（ＶＥＧＦ）及其受体（ＫＤＲ０在卵巢癌中的表达,并探讨其与卵巢癌发生的关系。方法：采用ＳＡＢＣ免疫组化染色法,对６６例卵巢肿瘤中,ＶＥＧＦ和ＫＤＲ的表达进行检测。结果：恶性,交界性及良性卵巢肿瘤中,ＶＥＧＦ和ＫＤＲ的表达率分别为７２．９％,７５．００Ａ％,３８．４６及５４．０５％,４３．７５％〈７．６９％;恶性肿瘤及交界性肿瘤ＶＥＧＦ和ＫＤＲ的表达,明显高于良性肿瘤。     

血管内皮生长因子及其受体在卵巢癌中表达的免疫组化检测

目的： 检测血管内皮生长因子（ＶＥＧＦ） 及其受体（ＫＤＲ） 在卵巢癌中的表达, 并探讨其与卵巢癌发生的关系。方法： 采用ＳＡＢＣ免疫组化染色法, 对６６ 例卵巢肿瘤中, ＶＥＧＦ 和ＫＤＲ 的表达进行检测。结果： 恶性、交界性及良性卵巢肿瘤中, ＶＥＧＦ 和ＫＤＲ 的表达率分别为７２－９％ 、７５－００ ％ 、３８－４６ ％ 及５４－０５％ 、４３－７５％ 、７－６９ ％ ; 恶性肿瘤及交界性肿瘤ＶＥＧＦ和ＫＤＲ的表达, 明显高于良性肿瘤（Ｐ＜ ０－０５） 。ＫＤＲ 不仅表达于肿瘤血管内皮细胞, 在肿瘤细胞内也有强表达。有淋巴结转移的卵巢癌ＶＥＧＦ 的表达与无淋巴结转移者相比较, 有显著差异（Ｐ＜ ０－０５）; ＫＤＲ 的表达与卵巢癌淋巴结转移无关（Ｐ＞ ０－０５）; 卵巢癌的不同临床分期、病理类型及病理分级间ＶＥＧＦ和ＫＤＲ的表达, 无显著性差异（ Ｐ＞ ０－０５） 。结论： ＶＥＧＦ和ＫＤＲ 的表达可能与卵巢癌的发生有关。     

去甲二氢愈创木酸对胶质瘤细胞诱导的内皮细胞迁移的影响

目的 观察去甲二氢愈创木酸（ＮＤＧＡ）对胶质瘤细胞诱导的内皮细胞迁移的影响。方法 采用微孔滤膜培养小室及室细胞联合培养法,进行人脐静脉内皮细胞系ＥＣＶ－３０４细胞与人恶性胶质瘤细胞系ＳＨＧ－４４细胞的联合培养,并以免疫组化ＳＰ方法检测ＳＨＧ－４４细胞血管内皮生长因子（ＶＥＧＦ）、碱性成纤维细胞生长因子（ｂＦＧＦ）的表达。结果 １００μｍｏｌ／Ｌ的ＮＤＧＡ作用１～３ｄ后,ＳＨＧ－４４细胞ＶＥＧＦ、ｂ     

活动类风湿关节炎病人血清ＶＥＧＦ水平测定

目的：为了解血管内皮细胞长生因子（ＶＥＧＦ）在类风湿关节炎（ＲＡ）病人中的水平。方法：ＩＬＩＳＡ双抗夹心法ＶＥＧＦ试剂合定量测定,同时采用ＢeckmanArray360检测血清ＲＦ水平,结果：３２例活动性ＲＡ病人血清ＶＥＧＦ水平健康对照组有明显增高（Ｐ＜０．０１）,结论：提示ＶＥＧＦ参与了ＲＡ的发病,测定血清ＶＥＧＦ水平是一种无侵入性,实用的监测ＲＡ病情的方法。     

缺血代氧对大鼠心肌产生血管内皮生长因子的影响及？…

目的：了解缺血低氧刺激与大鼠心肌血管内皮生长因子（ＶＥＧＦ）产生的关系、意义及其可能机制。方法：１．建立Ｗｉｓｔａｒ大鼠急性心肌梗塞模型,将２８只大鼠随机分为４组,每组７只：Ａ组正常对照;Ｂ组急性心肌梗１ｄ;Ｃ组急性心肌梗３ｄ;Ｄ组急性心肌梗７ｄ;Ａ组正常对照;Ｂ组急性心肌梗塞１ｄ;Ｃ组急性心肌塞７ｄ。心甩冰冻切片,免疫组化探测ＶＥＧＦ。２．原代培养大鼠心肌细胞随机分组：单纯缺氧组,Ａ：缺氧培养０     

血管内皮生长因子基因的表达及血管生成作用

构建新的血管内皮生长因子（ＶＥＧＦ）高效真核表达载体ｐｃＤ２／ＶＥＧＦ,体外转染ＶＳＭＣ〈发现其在ＶＳＭＣ中的表达效率较ｐｃＤＮＡ３／ＶＥＧＦ明显增高,体外和体内实验证实ＶＥＧＦ基因具有显著促进血管生成的作用。     

卡托普利对SHR心肌VEGF表达及微血管的影响

目的检测卡托普利降压治疗前后自发性高血压(SHR)心肌血管内皮生长因子(VEGF)表达水平与微血管密度的变化,探讨该药物是否具有逆转微血管稀少的作用。方法60只8月龄大鼠均分成正常血压对照组(WKY),SHR组和卡托普利治疗SHR组,用单宁酸-氯化铁组织化学法检测心肌微血管,用免疫组化SP法检测心肌VEGF蛋白表达,并对上述3组心脏切片用计算机图象分析系统进行定量观察分析。结果对照组SHR心肌微血管密度比对照组WKY减少,而VEGF表达水平比WKY组增强(均P<0.05),卡托普利治疗组SHR心肌微血管密度增加,而VEGF表达水平下降,与对照组SHR比较差异具有显著性(P<0.05),而与WKY组相当(P>0.05)。结论卡托普利在降压治疗的同时可下调VEGF表达水平,并逆转微血管减少,从而对靶器官具有保护作用。     

短链酰基辅酶A脱氢酶在大鼠心脏发育中的表达及其与心肌肥厚的关系

 目的：研究大鼠心脏发育过程中短链酰基辅酶A脱氢酶(short-chain acyl-CoA dehydrogenase, SCAD)的表达变化规律,并探讨其与高血压大鼠心肌肥厚的关系。方法：观察不同时期Wistar大鼠和不同周龄自发性高血压大鼠心肌组织的SCAD蛋白表达及酶活性变化,检测大鼠的血清和心肌游离脂肪酸含量。结果：与胚胎期19 d Wistar大鼠组比较,出生后1 d、2周、6周及16周龄Wistar大鼠组心肌的SCAD蛋白表达及酶活性增加,血清和心肌游离脂肪酸含量明显减少,二者之间呈负相关,其中,从2周龄Wistar大鼠组开始差异有统计学意义。与周龄匹配的WKY大鼠组比较,2周龄自发性高血压大鼠组收缩压尚未升高,6周龄及16周龄自发性高血压大鼠组收缩压显著增高;各时点自发性高血压大鼠组的左室重量指数均明显增高,提示自发性高血压大鼠在血压升高之前,已经发生了明显的心肌肥厚。与周龄匹配的WKY大鼠组比较,2周、6周及16周龄自发性高血压大鼠组心肌的SCAD蛋白表达及酶活性明显下降,血清和心肌游离脂肪酸含量明显增加,呈显著负相关。结论：(1)SCAD蛋白表达随大鼠心脏的生长发育逐渐上调,可能与心脏对脂肪酸的利用增加密切相关。(2)SCAD的蛋白表达及其酶活性显著下降, 可能是导致自发性高血压大鼠肥厚心肌能量代谢“胚胎型再演”的分子基础。     

VEGF在自发性高血压大鼠肾小球内的表达

目的研究血管内皮生长因子（VEGF）在幼年及成年自发性高血压大鼠（SHR）肾小球内的表达,探讨它在高血压病肾损害中的作用。方法6周龄SHR和WKY大鼠各10只,15月龄SHR和WKY大鼠各20只,将肾组织进行常规病理和免疫组化染色（SP法）,利用计算机图像分析定量肾组织内的微血管数和VEGF的表达。结果VEGF蛋白主要分布于肾小球内的足细胞和球内系膜细胞,15月龄SHR组的阳性反应程度显著高于其余3组（P〈0．05）,而其他各组间无显著差异。结论推测VEGF可能参与高血压肾损害的发病过程。     

Temporal characteristics of cardiomyocyte hypertrophy in the spontaneously hypertensive rat.

BACKGROUND: The spontaneously hypertensive rat (SHR) is frequently used as model of cardiovascular disease, with considerable disparity in reported parameters of hypertrophy. The aim of this study was to assess the temporal changes occurring during the development and progression of cardiomyocyte hypertrophy in SHR, subsequent to pressure overload, compared to changes associated with normal aging using the normotensive Wistar-Kyoto (WKY) rat. METHODS: Ventricular cardiomyocytes were isolated from rats at 8, 12, 16, 20 and 24 weeks, and parameters of hypertrophy (cell dimensions, protein mass, de novo protein synthesis, and gene expression) and function (contraction and hypertrophic responsiveness in vitro) were assessed. RESULTS: Hypertension was evident at > or =7 weeks in SHRs. Heart:body mass ratio, cardiomyocyte protein mass and width were elevated (P<.05) in SHRs at 16-20 weeks compared to WKYs. In SHRs compared to WKYs at 16 weeks, there was a transient increase (P<.05) in protein synthesis, enhanced hypertrophic responsiveness to phorbol-12-myristate-13-acetate, and induced hypertrophic responsiveness to isoprenaline. Skeletal-alpha-actin mRNA was detected in SHR but not WKY cells at all ages. ANP mRNA was lower in SHR than in WKY cells at 8-20, but progressively increased (P<.05) from 12 to 24 weeks within SHRs. Contractile function increased (P<.05) at 20 weeks in SHR compared to WKY rats. CONCLUSION: Structural and functional changes occurring at the cellular level in the myocardium of SHR follow a distinct pattern, such that pressure overload was initially accompanied by expressional changes (8-12 weeks), followed by active hypertrophic growth and enhanced function (16-20 weeks), which subsequently decelerated as stable compensation was attained.     

阿托伐他汀对自发性高血压大鼠心肌组织PPARs表达的影响

目的： 探讨阿托伐他汀对自发性高血压大鼠心肌组织PPARs（peroxisome proliferator-activated receptors, PPARs）表达的影响及其对心肌肥厚的逆转作用与可能机制。方法: 自发性高血压大鼠分为阿托伐他汀灌胃治疗组（SHR-A，30 mg·kg-1·d-1）及模型组（SHR），治疗8周，同周龄Wistar-Kyoto 鼠为正常血压对照组。治疗前及治疗后2、4、8周测量大鼠尾动脉血压。治疗后测血浆血脂水平，以心脏组织病理分析判断心肌肥厚，Western blotting 检测心肌组织PPARα、PPARγ的表达水平。结果: 经过8周治疗， SHR-A组及SHR组血压及血脂水平无明显差异（P＞0.05）。SHR-A组左室重量指数低于SHR组（P＜0.01）。在SHR-A组，PPARα及PPARγ表达高于SHR组（P＜0.01）。结论: 阿托伐他汀显著改善自发性高血压大鼠心肌组织PPARs表达，有效逆转左室肥厚，可能与其降压及降脂作用无关。     

上调miR-133a表达水平对自发性高血压大鼠心肌纤维化的影响

目的:观察上调微小RNA-133a(miR-133a)的表达水平对自发性高血压大鼠(SHR)心肌纤维化的影响。方法:以同源正常血压Wistar-Kyoto(WKY)大鼠为正常对照组,另将SHR随机分为SHR组、SHR+腺相关病毒(AAV)组和SHR+携带miR-133a的腺相关病毒(miR-133a-AAV)组。通过冠脉灌注法将miR-133a-AAV转染至SHR大鼠的心脏,监测大鼠的尾动脉压,Masson染色观察心肌胶原沉积情况,real-time PCR检测心肌组织中miR-133a的表达水平,免疫组化法和Western blot法检测心肌组织中转化生长因子-β1(TGF-β1)和结缔组织生长因子(CTGF)的蛋白表达水平。结果:与WKY大鼠相比,SHR的尾动脉压明显升高,心肌组织中miR133a表达水平降低,TGF-β1和CTGF蛋白表达水平升高,出现心肌纤维化;上调SHR心肌miR-133a的表达水平后,心肌纤维化程度明显减轻,TGF-β1和CTGF蛋白表达水平降低。结论:上调心肌组织中miR-133a的表达水平,对高血压导致的大鼠心肌纤维化有改善作用,其机制可能与抑制心肌组织中TGF-β1和CTGF蛋白表达有关。     

Myocardial cell growth and blood pressure development in neonatal spontaneously hypertensive rats

We evaluated the changing morphologic features of cardiac muscle cells (myocytes) and nuclei from neonatal spontaneously hypertensive rats (SHR) and their parent, normotensive strain Wistar Kyoto rats (WKY) and compared these with increasing heart weight and blood pressure development to determine if alterations in cell growth were present at this early stage of development. Femoral artery blood pressures were obtained from rats at 2 to 5-day intervals from birth to 21 days of age by using a micropipette servo-null pressure recording system. Tritiated thymidine autoradiography was used to study myocyte nuclear development, and isolated myocytes were prepared to evaluate cell-size changes by using a Coulter Counter system (Coulter Electronics, Hialeah, Florida). Heart weight and blood pressure were elevated in SHR when compared to WKY at birth. Myocytes were all mononucleated at birth in both strains and were of equal size, demonstrating that the larger heart mass in SHR was due to an increased number of cells. Heart weight relative to body weight remained greater in SHR when compared to WKY throughout the 28-day study period, but cell numbers became equal in the two strains by the 2nd week. A this time (6 to 9 days postpartum) blood pressure was also similar in both strains, but increased significantly again in SHR by 15 and 21 days of age. Cell maturation occurred earlier in SHR than in WKY as indicated by an earlier development of binucleate myocytes and there was an earlier initiation of hypertrophic myocyte growth in SHR. Increase in SHR cell size occurred at a time when blood pressures were not different, suggesting that greater cell size in SHR than in WKY was not due to differences in blood pressure. Therefore, when compared to the WKY, the SHR had three phases of altered cell growth: a first phase of accelerated hyperplastic growth during the fetal period, and a second phase (6 to 12 days of age) of earlier initiation of hypertrophic cell growth and increased myocyte size. The SHR myocyte changes in the second phase occurred while the SHR and the WKY blood pressures were not significantly different. Finally, in a third phase (at 15 days of age and over), SHR had a sustained increase in myocyte size as well as elevated blood pressure.     

The transverse tubular system of the hypertrophie myocardium: morphology and morphometry in spontaneous hypertensive rats (SHR)

Summary We reported previously on a modified Golgi stain that, in conjunction with high voltage electron microscope stereoscopy, gives striking views of the elaborate network of the transverse tubular system (T system) in rat myocardium. In this report we used the same techniques to study three-dimensional arrangements of the T system in the left ventricular myocardium of spontaneous hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY). High voltage electron microscope stereoscopy revealed distinctive morphological characteristics of the T system, such as undulating running, short dead-end branches, and labyrinth-like tubular aggregates in the hypertrophic myocardium of SHR. Quantitative analysis of the SHR T system indicated a surface area greater than that of WKY. These findings may support the hypothesis that making an additional T system membrane will compensate for the smaller surface-to-volume ratio. However, the normal regulatory mechanism required to maintain the surface-to-volume ratio does not function properly in SHR, resulting in morphological abnormalities and functional disturbances of the myocardium.     

Differential expression of α-enolase in the normal and pathological cardiac growth

Objectives

It was found that α-enolase was dramatically up-regulated in the hypertrophic hearts of SHR in our previous study. The purposes of this study were to examine the expression pattern of α-enolase in pre- and postnatal myocardium of spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats, and to explore the relationship between the overexpression of α-enolase and left ventricular hypertrophy.

Methods

HE staining was used for the measurement of cardiac hypertrophy. Immunohistochemical technique was used to evaluate the location of α-enolase. The expressions of α-enolase in the left cardiac ventricles at different development times were examined by Real-time RT-PCR and Western blot.

Results

Cardiac hypertrophy was found in SHR rats at 4 weeks of age and remained up to 24 weeks of age. The signals of α-enolase protein were strong and existed extensively in hypertrophic myocardium in SHR, while in the normal myocardium of WKY, the signals were scarcely found and weak. The levels of α-enolase mRNA and protein in SHR and WKY hearts during fetal stage and newborn stage were similar, while from 4 weeks of age to 24 weeks of age, accompanied by the cardiac hypertrophy, the levels of α-enolase mRNA and protein in left ventricle of SHR were significantly higher than that in WKY.

Conclusions

The expressions of α-enolase in the left ventricle of the rats during normal and pathological cardiac development were different. This phenomenon provides the potential clues to understanding pathophysiological mechanisms in cardiac hypertrophy of SHR.     

SHR大鼠血压升高前后脑内VEGF的变化及其意义

为探讨自发性高血压大鼠(SHR)血压升高前后脑内血管内皮生长因子(VEGF)表达水平的变化及其意义,本研究应用免疫组化SP法及计算机图像分析技术,对幼年(6周龄)和成年(12月龄)SHR大鼠及幼年(6周龄)和成年(12个月龄)同品系正常血压对照组京都Wistar大鼠(WKY)脑切片VEGF蛋白的表达水平进行了定量分析。结果发现在4组大鼠大脑皮质的广泛区域、纹状体、室管膜上皮及脉络丛等处均观察到散在分布的VEGF样免疫阳性细胞,阳性反应产物呈棕色颗粒状,位于胞浆中,其中幼年和成年WKY及幼年SHR三组间阳性细胞数及其灰度差异无显著性(P>0.05),而12月龄SHR组阳性细胞数及其灰度均高于其它三组(P<0.05)。以上结果提示:随着成年SHR大鼠血压的升高,脑内VEGF蛋白的表达水平上调,高表达的VEGF可能对高血压时脑缺血缺氧诱导的血管内皮细胞凋亡及微血管的存活有保护意义,同时也可能参与了高血压所致的脑损伤。