SHR和WKY大鼠主动脉hsp70mRNA水平的比较研究

本工作应用热激蛋白７０ＫＤ（ｈｓｐ７０）核酸分子杂交方法，检测了：（１）自发性高血压（ＳＨＲ）主动脉和离休培养的主动脉平滑肌细胞受热激后ｈｓｐ７０ｍＲＮＡ水平的变化；（２）不同细胞培养时间（３个月与６周）的ＳＨＲ、ＷＫＹ主动脉ｈｓｐ７０ｍＲＮＡ水平。结果提示ＳＨＲ主动脉ｈｓｐｍＲＮＡ水平增加，ＳＨＲ细胞培养受热激（４２℃，１５ｍｉｎ）后２ｈ，ｈｓｐ７０ｍＲＮＡ水平明显高于ＷＫＹ鼠者，６周较３个月的ＳＨＲ细胞ｈｓｐ７０ｍＲＮＡ水平高；６个月的ＳＨＲ细胞较同期和３个月的ＷＫＹ细胞ｈｓｐ７０ｍＲＮＡ高。推论ＳＨＲ血管平滑肌细胞对热敏感，原癌基因ｃ－ｍｙｃ和抗癌基因Ｐ５３可能参与ｈｓｐ７０表达调控，并共同参与ＳＨＲ细胞增殖的调节。     

佛波脂及Microcystine-LR对SHRsp内脏阻力血管平滑肌钙通道的影响

目的 研究佛波脂及ｍｉｃｒｏｃｙｓｔｉｎｅ－ＬＲＭＣＹＳＴ－ＬＲ）对卒中易感型自发性高血压大鼠（ＳＨＲｓｐ）及常压对照Ｗｉｓｔａｒ大鼠肠纱膜动脉Ａ４－Ａ５段分支阻力血管平滑肌电压依赖性钙通道的影响。方法 采用膜片箍全细胞钡电流方式记录钙通道的活力。结果 佛波脂激活蛋白激酶Ｃ（ＰＫＣ）或ＭＣＹＳＴ－ＬＲ换制细胞蛋白磷酸酶（ＰＰＩＡ、ＰＰⅡＡ）而相对增加磷酸化过程,均可激活内脏阻力血管平滑肌电压依赖性     

不同年龄自发性高血压大鼠血管平滑肌细胞Ca^2＋转运功…

应用不同年龄自发性高血压大鼠（ＳＨＲ）血管平滑肌原代与传代细胞（ＶＳＭＣ），观察Ｃａ＾２＋转运功能障碍及某些相关因素的变化，结果表明：（１）在血压未升高的ＶＳＭＣＣａ＾２＋内流已较同龄对照组ＷＫＹ大鼠明显增加，而Ｃａ＾２＋外流量较ＷＫＹ大鼠显著降低。表明ＳＨＲＶＳＭＣ膜Ｃａ＾２＋转运功能障碍在血压升高前就已发生，并随年龄及血压增高有加重趋势；（２）ＶＳＭＣｃＡＭＰ及钙调素含量变化与细胞膜Ｃａ＾２＋     

知管紧张素Ⅱ及其拮抗剂对纱膜细胞内游离钙的影响

目的 探讨Ｌｏｓａｒｔａｎ拮抗局部肾素－血管紧张系（ＲＡＳ）对预防高血压致肾脏靶器官损伤的意义,方法 应用细胞培养和Ｆｕｒａ－２方法,观察血管紧张素Ⅱ和血管紧张素Ⅱ型受体（ＡＴ１受体）拮抗剂Ｌｏｓａｒｔａｎ对自发性高血压大鼠（ＳＨＲ）和正常血压大鼠（ＷＫＹ）肾小球系膜细胞内游离钙浓度的影响。结果 ＡｎｇⅡ使ＳＨＲ系膜细胞「Ｃａ＾２＋」ｉ增高,对ＷＫＹ无影响;ＡｎｇⅡ对ＳＨＲ系膜细胞「Ｃａ＾２＋」ｕ     

高血压高胆固醇血症大鼠对血管收缩性刺激的反应

研究高血压合并高胆固醇血症大鼠血管对去甲肾上腺素（ＮＥ），ＫＣｌ和ＣａＣｌ２收缩性刺激的运动反应。１２周龄的自发性高血压大鼠（ＳＨＲ）和京都种Ｗｉｓｔａｒ大鼠（ＷＫＹ）分成４组：胆固醇喂养的ＷＫＹ和ＳＨＲ组及其对照组。测定血压、心／体比和血脂。胆固醇喂养１２周后，ＷＫＹ和ＳＨＲ的血压没有明显变化；但血浆胆固醇和甘油三酯增加，ＨＤＬ降低。胆固醇组ＳＨＲ的心／体比降低。主动脉条未发现肉眼可见的动脉粥样硬化缺损。高胆固醇血症的ＷＫＹ和ＳＨＲ的主动脉条ＮＥ量效曲线显著左移，ＥＤ５０降低，ＳＨＲ较ＷＫＹ更显著。胆固醇组ＷＫＹ和ＳＨＲ的主动脉条ＫＣｌ和ＣａＣｌ２量效曲线右移，ＥＤ５０增高。高血压增强血管对ＮＥ的反应，特别是并发高胆固醇血症大鼠的血管。     

不同年龄自发性高血压大鼠血管平滑肌细胞Ca~（2＋）转运功能障碍及某些相关因素的变化

应用不同年龄自发性高血压大鼠（ＳＨＲ）血管平滑肌原代与传代细胞（ＶＳＭＣ），观察Ｃａ２＋转运功能障碍及某些相关因素的变化。结果表明：（１）在血压未升高的ＶＳＭＣＣａ２＋内流已较同龄对照组ＷＫＹ大鼠明显增加，而Ｃａ２＋外流量较ＷＫＹ大鼠显著降低。表明ＳＨＲＶＳＭＣ膜Ｃａ２＋转运功能障碍在血压升高前就已发生，并随年龄及血压增高有加重趋势；（２）ＶＳＭＣｃＡＭＰ及钙调素（ＣａＭ）含量变化与细胞膜Ｃａ２＋转运功能障碍基本同步，提示二者异常与细胞膜Ｃａ２＋转运功能障碍有密切关系；（３）血压升高前ＳＨＲＶＳＭＣ内ＡＮＧⅡ含量与ＷＫＹ大鼠相比无明显差异，但１６周龄５ＨＲＶＳＭＣ内ＡＮＧⅡ含量明显高于其幼鼠（Ｐ＜０．００１），与血压呈正相关。提示ＶＳＭＣ内ＡＮＧⅡ在血压升高过程中可能具有一定作用。以上结果表明高血压时ＶＳＭＣ膜Ｃａ２＋转运功能障碍有明显遗传倾向，ＶＳＭＣ内ＣＡＭＰ、ＣａＭ及ＡＮＧⅡ含量异常与上述障碍密切相关。     

SHR和WKY大鼠主动脉hsp 70 mRNA水平的比较研究

本工作应用热激蛋白７０ＫＤ核酸分子杂交方法，检测了：（１）自发性高血压主动脉和离体培养的主动脉平滑肌细胞受热激后ｈｓｐ ７０ｍＲＮＡ水平的变化；（２）不同细胞培养时间的ＳＨＲ、ＷＫＹ主动脉ｈｓｐ ７０ｍＲＮＡ水平。结果提示ＳＨＲ主动脉ｈｓｐ ｍＲＮＡ水平增加，ＳＨＲ细胞培养受热激后２ｈ，ｈｓｐ ７０ｍＲＮＡ水平明显高于ＷＫＹ鼠者，６周较３个月的ＳＨＲ细胞ｈｓｐ ７０ｍＲＮＡ水平高；６个月的ＳＨＲ细     

自发性高血压大鼠主动脉血管平滑肌细胞异常增殖和自身肾素－血管紧张素系统的关系

目的以２０周龄自发性高血压大鼠（ＳＨＲ）和正常血压大鼠（ＷＫＹ）胸主动脉平滑肌细胞（ＡＳＭＣ）为模型，探讨ＳＨＲＡＳＭＣ异常增殖和自身肾素－血管紧张素系统（ＲＡＳ）的关系。方法用３Ｈ－ＴｄＲ参入量和倍增时间（ＤＴ）反映ＡＳＭＣ的增殖能力，放射免疫法测定血管紧张素Ⅱ（ＡｎｇⅡ）浓度，紫外分光光度法测定血管紧张素转换酶（ＡＣＥ）活性。结果（１）ＳＨＲＡＳＭＣ３Ｈ－ＴｄＲ参入量显著高于ＷＫＹ，ＤＴ显著短于ＷＫＹ（Ｐ＜０．０１）。基础状态下，ＳＨＲＡＳＭＣ合成ＡｎｇⅡ、ＡＣＥ以及分泌ＡｎｇⅡ的量显著高于ＷＫＹ（Ｐ＜０．０１）。（２）在含２％ＦＣＳ的培养基中，１０－６ｍｏｌ／ＬＡｎｇⅡ使ＳＨＲ、ＷＫＹ的３Ｈ－ＴｄＲ参入量分别提高到对照组的３．３倍、２．２倍（Ｐ＜０．０１），ＳＨＲＡＳＭＣ在不同浓度ＡｎｇⅡ刺激时的３Ｈ－ＴｄＲ参入量显著高于ＷＫＹ（Ｐ＜０．０１），１０－６ｍｏｌ／ＬＡｎｇⅡ使ＳＨＲＡＳＭＣ细胞数增加７２．５±２３．１％（Ｐ＜０．０１），ＷＫＹＡＳＭＣ细胞数无显著增加，１０倍浓度ＡｎｇⅡ的Ｓａｒａｌａｓｉｎ对ＳＨＲ、ＷＫＹ３Ｈ－ＴｄＲ参入的抑制率分别为６６．７±３．３％，４４．７±９．９％（Ｐ＜０．０１）     

大鼠自发性高血压和心肌肥厚时热休克蛋白70基因表达的研究

本工作采用热休克蛋白７０（ＨＳＰ７０）核酸分子杂交方法，检测了自发性高血压大鼠（ＳＨＲ）与其正常对照（ＷＫＹ）大鼠离体培养的主动脉平滑肌细胞（ＡＳＭＣ）受热刺激后以及大鼠腹主动脉缩窄后心肌肥厚时左心室ＨＳＰ７０ｍＲＮＡ的水平，并对ＳＨＲ和ＷＫＹ肝组织基因组ＤＮＡ进行了限制性酶切片断长度多态性（ＲＦＬＰ）分析。结果表明：ａ．３７℃培养的ＳＨＲＡＳＭＣ及整体ＳＨＲ肝组织ＨＳＰ７０ｍＲＮＡ的基础表达水平均低于ＷＫＹ大鼠，ＳＨＲＡＳＭＣ受热刺激（４２℃１５ｍｉｎ）后２ｈ，ＨＳＰ７０ｍＲＮＡ表达增加的程度明显大于ＷＫＹ大鼠。ｂ．大鼠腹主动脉缩后造成左室压力超负荷，左心室ＨＳＰ７０ｍＲＮＡ在压力超负荷４ｈ时已明显升高，１ｄ、２ｄ、１ｗ均维持在高水平，１ｗ后逐渐降低。ｃ．ＳＨＲ和ＷＫＹ鼠肝组织基因组ＤＮＡ经ＢａｍＨＩ酶切后，ＳＨＲＨＳＰ７０基因缺失一条约５．６ｋｂ的片段。提示：ＳＨＲＡＳＭＣ对热敏感性增加；压力负荷增加早期，左心室ＨＳＰ７０ｍＲＮＡ表达明显增加；ＨＳＰ７０基因结构异常可能与高血压发生有关。     

WKY大鼠与SHR血管平滑肌神经肽Y受体数量及其亲和力的对照研究

目的：了解Ｗｉｓｔａｒ－Ｋｙｏｔｏ大鼠（Ｗｉｓｔａｒ－Ｋｙｏｔｏｒａｔｓ,ＷＫＹ）、自发性高血压大鼠（Ｓｐｏｎｔａｎｅｏｕｓｌｙｈｙｐｅｒｔｅｎｓｉｖｅｒａｔｓ,ＳＨＲ）血管平滑肌细胞（Ｖａｓｃｕｌａｒｓｍｏｏｔｈｍｕｓｃｌｅｃｅｌｓ,ＶＳＭＣ）上神经肽Ｙ（ＮｅｕｒｏｐｅｐｔｉｄｅＹ,ＮＰＹ）受体的表达水平及其差异。方法：应用Ｉｏｄｏｇｅｎ法对ＮＰＹ受体进行了测定。结果：ＮＰＹ蛋白质标记率为７０％,比活度为３．３６ＭＢｑ／μｇＶＳＭＣ在（１．０～５．０）×１０６细胞／ｍｌ的范围内,１２５Ｉ－ＮＰＹ的特异性结合量与ＶＳＭＣ细胞数之间呈线性关系;ＳＨＲ的ＮＰＹ受体最大结合量（Ｂｍａｘ）和受体复合物解离常数（Ｋｄ）均明显高于ＷＫＹ,相关分析显示Ｂｍａｘ,Ｋｄ与血压增高程度呈正相关。结论：ＮＰＹ受体数目和亲和力的增加可能在高血压的发病机制中占有重要的地位     

Atrial natriuretic factor and renin synthesized in cultured aortic smooth muscle cells of rats.

This study was designed to determine whether or not atrial natriuretic factor (ANF) is present in the vascular walls and to observe the differences in ANF between control (WKY) and stroke-prone spontaneously hypertensive rats (SHRsp). It was found that ANF is indeed present in the vascular wall of the distal aorta. HPLC analysis of the extracts from cultured aortic smooth muscle cells (ASMC) and medium revealed that intracellular ANF was mainly in the form of ANF(1-126), at levels of 0.82 +/- 0.03 (SHRsp) and 1.04 +/- 0.10 ng/10(6) cells (WKY), while the major form in the medium was ANF(99-126), at levels of 0.40 +/- 0.06 and 0.60 +/- 0.06 ng/10(6) cells, respectively. Both forms were present in smaller amounts in SHRsp than in WKY rats. On the contrary, both renin activity and angiotensin I concentrations in SHRsp cells were significantly higher than those in the WKY controls. In addition, immunocytochemistry showed positive ANF staining in cultured ASMC of both strains. The results suggest that ANF can be synthesized and secreted by cultured ASMC from rats.     

抗高血压因子对正常及高血压动物动脉平滑肌Ca~(2+)内流的影响

通过实验观察到从自发性高血压大鼠(SHR)红细胞提取的抗高血压因子(AHF)可以明显抑制SHR血管平滑肌(VSM)Ca~(2+)内流,呈剂量依赖关系。而对正常血压大鼠(WKY)无影响。这种抑制作用及与剂量的关系在肠系膜动脉比主动脉更明显。结果表明,AHF的降压作用可能与抑制VSM Ca~(2+)内流有关。     

抗高血压因子对正常及高血压动物动脉平滑肌C...

Aorta segments (A) and mesenteric arteries (MA) from stroke prone spontaneously hypertensive rats (SHRsp) and control Wistar Kyoto rats (WKY) were used in the present study to assess the effect of AHF on Ca2+ influx in vascular smooth muscle (VSM). The results indicated that Ca2+ influx in VSM of SHRsp was much higher than that of WKY rats (P less than 0.05). AHF at 10(-7), 10(-6) and 10(-5) g/ml can significantly inhibit Ca2+ influx in a dose-dependent manner in VSM of both A and MA (P less than 0.05 and less than 0.01). The suppression effect of AHF on Ca2+ influx and the concentration-dependent relationship were more obvious in MA than in A. The Ca2+ influx in VSM of WKY rats was unaffected by administration of AHF.     

Changes of Ca^2＋ activated potassium channels and cellular proliferation in autoge-nous vein grafts

Objective: To investigate changes of Ca2+ activated potassium channels ( KCa) in autogenous vein grafts. Methods: Contraction of venous ring was measured by means of perfusion in vitro. The intimal rabbits proliferation of vascular and proliferation of cultured smooth muscle cells ( vascular smooth muscle cells, VSMCs) were observed by the means of computerised image analysis and MTT method respectively. Furthermore, whole cell mode of patch clamp was used to record KCa of VSMCs isolated from autogenous vein grafts. Results: One week after transplantation there were no significant differences of contraction and intimal relative thickness between autogenous vein grafts and control. Contraction and intimal relative thickness of autogenous vein graft were significantly increased 2 weeks after transplantation (P < 0. 05 , n = 8 vs control) , and they was more enhanced 4 weeks after vein transplantation ( P < 0. 01 , n =8 vs control ). TEA( blocker of Ca2+ activated potassium channels) increased MTT A490 nm     

牛磺酸对大鼠离体气管平滑肌舒张作用及机制的研究

目的研究牛磺酸（Tau）对大鼠离体气管平滑肌的作用及可能机制。方法观察牛磺酸对高钾预收缩大鼠离体气管环的作用,通过工具药观察牛磺酸对气管环作用的可能机制。经Powedab生物信号测定系统记录气管环张力。结果①牛磺酸【（10～80）mmol／L】对高钾（30mmol／L）诱发的大鼠离体气管环收缩有舒张作用,且呈浓度依赖性,最大舒张率为（50．22±18．30）％。②牛磺酸舒张大鼠气管平滑肌的作用可被K、Ca通道抑制剂四乙胺（10mmol／L）和KATP通道抑制剂格列苯脲（10μmol／L）显著减弱,而KIR通道抑制剂氯化钡（1mmol／L）和KV通道抑制剂4-氨基吡啶（1mmol／L）对此则无显著影响。（3）L—NAME（1mmol／L）、Pino（10μmol／L）、Indo（10μmol／L）对牛磺酸舒张大鼠离体气管环无明显影响。结论牛磺酸可舒张大鼠离体气管平滑肌．且舒张作用呈浓度依赖性。其舒张作用与一氧化氮、前列环素和β受体无关,可能与激活KATP通道和K、Ca通道有关。     

黄芪对SHR心肌SERCA表达及血管平滑肌钙离子水平影响的初步探讨

目的:探讨黄芪注射液对SHR心肌SERCA的表达及血管平滑肌钙离子含量的影响.方法:将19只自发性高血压大鼠(SHR)与16只Wistar-kyoto大鼠(WKY)随机各分为空白对照组、低黄芪剂量组、中黄芪剂量组、高黄芪剂量组四组,分别每天腹腔注射0.9ml生理盐水和0.9ml、1.2ml、1.8ml的黄芪注射液共8周,再测量大鼠心肌中SERCA的表达和血管平滑肌钙离子含量.结果:SHR生理盐水组心肌SERCA在蛋白水平的表达,较WKY生理盐水组明是下降;SHR大剂量黄芪组心肌SERCA在蛋白水平的表达,较SHR生理盐水组明显升高.SHR生理盐水组血管平滑肌钙离子水平明显高于WKY组:SHR大剂量黄芪组血管平滑肌钙离子水平较SHR生理盐水组明显降低.结论:大剂量黄芪注射液能显著增加SHR心肌SERCA在蛋白水平的表达,能明显的减少SHR血管平滑肌钙离子的水平.     

Lovastatin increases nitric oxide synthesis in IL- 1β- stimulated smooth muscle cells

Objective Nitric oxide (NO) production by inducible NO synthase (iNOS) may play an importa nt role in the pathogenesis of atherosclerosis. Although lovastatin has been s hown to reduce the progression of atherosclerosis, it is not known whether it re gulates NO production. We investigated the effects of lovastatin on NO synthesi s and the mechanisms by which lovastatin exerts its effects in rat vascular smoo th muscle cells. Methods Primary cultures of the vascular smooth muscle cells were obtained from the medi a of the thoracic aorta of Sprague Dawley rats (200-250 g). Nitrite levels in the culture medium of rat vascular smooth muscle cells were determined colorimet rically. Results Lovastatin (10(-5) mol/L) significantly increased interleukin- 1β (IL- 1β , 10 ng/mL)- induced nitrite accumulation in a time (0-24 hours)- dependent man ner. Exogenous mevalonate and geranylgeranyl- pyrophosphate completely reverse d the stimulatory effects of lovastatin on nitrite production. Furthermore, in hibition of Rho by C3 exoenzyme mimicked the increase in IL- 1β- induced nitrit e accumulation induced by lovastatin in the vascular smooth muscle cells. Conclusion These results demonstrate that lovastatin up- regulates NO formation in rat vasc ular smooth muscle cells stimulated by IL- 1β, and the effect may be associated with the inhibition of Rho activity.     

ET－1和NO对肺动脉平滑肌细胞DNA合成的作用及缺氧对其调制的研究

观察内皮素－１（ＥＴ－１）和一氧化氮（ＮＯ）在缺氧性肺血管结构重组中的作用，发现ＥＴ－１可促进肺动脉平滑肌细胞（ＰＡＳＭＣ）ＤＮＡ合成，其促进作用呈剂量依赖性；ＮＯ供应剂ＳＮＰ则起抑制作用，ＮＯ的抑制增殖作用主要由ｃＧＭＰ介导。在此基础上观察到缺氧可促进ＰＡＳＭＣ对ＥＴ－１的增殖反应，同时可抑制ＰＡＳＭＣ胞浆内可溶性鸟苷酸环化酶活性而降低ＰＡＳＭＣ对ＮＯ等舒血管药物的反应性。提示ＥＴ－１和ＮＯ及缺氧在缺氧性肺血管结构重组中具有重要的作用。