雌激素对血管平滑肌细胞凋亡的影响

目的 :观察雌激素对血管平滑肌细胞 (SMC)凋亡的影响。方法 :以大鼠为对象 ,观察正常对照组、去卵巢组及去卵巢 +雌二醇治疗组大鼠血管平滑肌细胞凋亡的关系。结果 :雌二醇治疗组血管平滑肌细胞凋亡较去卵巢组相比明显减少(P>0 .0 5 ) ,与正常对照组比较差别无显著性意义 (P >0 .0 5 )。结论 :雌二醇对血管平滑肌细胞的凋亡有抑制作用 ,可能是其预防动脉粥样硬化的机制     

血管平滑肌细胞生长的负调控

血管平滑肌细胞生长的负调控戴生明，单综述缪朝玉，苏定冯审校（第二军医大学药理学教研室，上海２００４３３）中国图书分类号Ｒ３９２．２；Ｒ９７２血管平滑肌细胞（ＶＳＭＣ）生长包括增殖（细胞数量的增加）和肥大（细胞体积的增加）。ＶＳＭＣ异常生长是高血压、动...     

氧化型低密度脂蛋白诱导血管平滑肌细胞凋亡

AIM: To examine whether oxidized low density lipoproteins (ox-LDL) could induce apoptosis in rabbit aortic smooth muscle cells (VSMC). METHODS: Low density lipoproteins (n-LDL) were isolated from healthy human plasma by gradient ultracentrifugation and oxidized by CuSO4 10 mumol.L-1. VSMC were exposed to ox-LDL, n-LDL, or phosphate-buffer solution (PBS) as control. Morphological changes were observed under fluorescene microscope after Hoechst 33258 staining. Extracted DNA was electrophoresized on agarose gel. RESULTS: Incubation of VSMC with ox-LDL 300 mg.L-1, not n-LDL, for 24 h induced morphological apoptosis changes (chromatin condensation, nucleus fragmentation) and DNA fragmentation, which was furthered with the incubation time up to 48 h or at a concentration of 400 mg.L-1. Dextran sulfate, a scavenger receptor blocker and butylated hydroxytoluene (BHT), an antioxidant, exhibited no effect on DNA fragmentation. Lysophosphatidylcholine (LPC) at a concentration up to 125 mumol.L-1 (equivalent to ox-LDL 300 mg.L-1) did not elicit DNA fragmentation. CONCLUSION: Ox-LDL induced apoptosis in VSMC without involving oxygen free radicals and LPC.     

他汀类药物与血管平滑肌细胞凋亡

血管平滑肌细胞（VSMC）的异常增殖和迁移在粥样斑块形成和冠状动脉介入治疗（PCI）后再狭窄中起非常重要的作用。他汀类药物不仅是一种有效的调脂药，同时还有多种调脂以外的作用。如他汀类药物可以抑制VSMC的增殖和迁移，并诱导其凋亡。临近病变部位的中层VSMC的凋亡可能造成斑块纤维帽容易破裂，进而引起斑块不稳定和临床事件。但研究证实他汀类药物有稳定斑块的作用，且新生内膜VSMC对他汀类药物诱导的凋亡作用比普通的血管中层VSMC更敏感。因此，他汀类药物的促新生内膜VSMC凋亡作用可能对预防PCI术后再狭窄起有益作用。具体作用机制以及临床上如何合理发挥此类作用尚待进一步研究。     

血管平滑肌细胞的增殖机制

血管平滑肌细胞 (ＶＳＭＣ)增殖是高血压、动脉粥样硬化和血管成形术后再狭窄等疾病中重要的病理改变[1 ] 。目前认为其发生机制与ＶＳＭＣ增殖的正负调控因子平衡失调密切相关[2 ] 。本文从ＶＳＭＣ增殖调控的角度对近年来ＶＳＭＣ增殖机制研究新的进展作一综述。1　细胞外调控ＶＳＭＣ增殖过程中 ,一些特定的细胞以旁分泌作用或靶细胞以自分泌作用产生ＶＳＭＣ生长及分化的调节物。分子生物学研究已揭示许多细胞外的生长因子、血管活性物质与增殖有关 ,以血小板源生长因子 (ＰＤＧＦ)、成纤维细胞生长因子 (ＦＧＦ)、转化生长因子 β(ＴＧ…     

Apelin抑制血管平滑肌细胞向成骨细胞分化

目的研究apelin对VSMCs向成骨细胞分化的作用以及作用机制。方法以体外培养的钙化型血管平滑肌细胞(CVMSCs)作为血管钙化的研究模型,通过检测碱性磷酸酶(ALP)活性以及骨钙素的分泌观察apelin与VSMCs向成骨细胞分化之间的关系,应用细胞外调节蛋白激酶(ERK)抑制剂、磷脂酰激醇3-激酶(PI3-K)抑制剂以及APJ的小干扰RNA(APJ siRNA)观察涉及的信号通路。结果Apelin抑制ALP活性、骨钙素分泌以及矿化结节的形成。CVSMCs表达APJ蛋白。Apelin激活ERK和蛋白激酶B(AKT,PI3-K的下游子)。应用siRNA沉默APJ取消了apelin对ERK和Akt活化的抑制作用。而且,抑制APJ表达、ERK或PI3-K的活化逆转了apelin对ALP活性的影响。结论 Apelin通过APJ/ERK和APJ/PI3-K/AKT信号途径抑制CVSMCs向成骨细胞分化。Apelin可能对动脉钙化有保护作用。     

大鼠血管平滑肌细胞的贴块法培养

目的:探讨大鼠血管平滑肌细胞的原代培养方法与生长特性,为中药研究提供实验材料。方法:采用组织贴块法培养,倒置显微镜观察形态,电镜鉴定,台盼蓝染色活力检测。结果:倒置显微镜和电镜观察培养的血管平滑肌细胞生长状态良好,呈现平滑肌细胞特征性的“峰—谷”状结构,台盼蓝染色约有95%以上的细胞为活细胞。结论:本培养方法简单、经济,经过一定时间的操作训练,即可成功培养出平滑肌细胞。     

血管平滑肌细胞粘附分子的表达与心血管疾病的关系

粘附分子是指由细胞合成、存在于细胞膜或细胞外,介导细胞间、细胞与细胞基质粘附的一大类分子。与心血管疾病有关的粘附分子包括整合素家族、免疫球蛋白超家族和选择素家族。免疫球蛋白超家族成员细胞间粘附分子（intercellular adhesion molecule-1,ICAM-1）和血管细胞粘附分子（vascular cell adhesionmolecule-1,VCAM-1）在白细胞向激活的内皮细胞粘附过程中起重要作用。血管内皮细胞表达粘附分子的作用已比较清楚,但几年前有人发现在动脉粥样硬化血管壁的内膜平滑肌细胞也表达ICAM-1和VCAM-1,这就引起了与这些分子相关问题的探讨。本文就ICAM-1和VCAM-1在血管平滑肌细胞表达的调节及在动脉     

缬沙坦对人脐静脉平滑肌细胞凋亡的影响

目的:研究血管紧张素受体阻滞剂缬沙坦对人脐静脉血管平滑肌细胞(VSMC)凋亡的调节作用,以探讨缬沙坦抗动脉粥样硬化的作用机制.方法:荧光显微镜观察缬沙坦作用后VSMC凋亡的形态学变化,用流式细胞仪测定缬沙坦用药前后VSMC凋亡率并作周期分析,用流式细胞仪检测缬沙坦对凋亡相关基因Bax,Bcl-2和caspase3表达的影响.结果:缬沙坦高浓度 血管紧张素Ⅱ(AngⅡ)组的VSMC凋亡率较AngⅡ模型组明显提高,缬沙坦单药高、中、低浓度组的VSMC凋亡率较对照组明显提高.细胞周期分析表明,AngⅡ 缬沙坦组与AngⅡ组比较,G0/G1期百分率显著升高,S期百分率显著降低,且具有浓度依赖性.Bax和caspase 3基因经高浓度缬沙坦组作用24 h后表达水平增加,Bcl-2则没有改变.结论:缬沙坦对AngⅡ诱导VSMC的增殖有抑制作用;缬沙坦可能促进VSMC凋亡;缬沙坦促进VSMC凋亡可能与Bax和caspase基因表达增加有关.     

Jak2在凝血酶诱血管平滑肌细胞增生中的作用

目的探讨凝血酶诱血管平滑肌细胞（vascular smooth muscle cells，VSMCs）异常增生的时相关系，及Jak2在其增生信号传递过程中的作用。方法本实验采用培养大鼠胸主动脉4—10代的VSMCs的单细胞悬液用于实验。用MTS法测VSMCs细胞增殖率。用免疫沉淀测定Jak2的活性。结果（1）0．5（u／ml）和1．0（u／ml）两种浓度的凝血酶相对于对照组，在各时间段均有明显促VSMCs增殖作用，且两者作用程度相似，增殖曲线呈双峰样改变，增殖峰在1h和24h。（2）凝血酶刺激组Jak2的活性在6h（P〈0．05）及12h（P〈0．05）均明显上升。相对于凝血酶组，AG490有效的降低了凝血酶诱导VSMCs1h峰值（P〈0．05）及24h峰值（P〈0．05）。结论（1）凝血酶促VSMCs的增殖在时间上呈双峰样改变，提示凝血酶不仅有早期快速促VSMCs的增殖作用．而且有后期促VSMCs的增殖作用。（2）由于Jak2活性高峰（6h及12h）在凝血酶促VSMCs第二增殖峰值（24h）之前。提示Jak2活性后期的增加参与了凝血酶促VSMCs后期增殖．     

Progenitor cell-derived smooth muscle cells in vascular disease

Accumulation of vascular smooth muscle cells (VSMCs) in the tunica intima plays a major role in the pathogenesis of atherosclerosis and restenosis following endovascular procedures. Arterial VSMCs are heterogeneous even in the normal vessel wall and display different phenotypes in physiological and pathological conditions. In the classical paradigm, vascular wall injury induces VSMC de-differentiation, proliferation and migration from the media into the intima in response to growth factors and proteolytic agents. Accordingly, VSMCs in atherosclerotic plaques and in restenosis display a de-differentiated or ‘synthetic’ phenotype compared to a ‘contractile’ phenotype in the normal media. In contrast, recent studies have identified bone marrow and peripheral blood-derived endothelial and VSMC progenitors that may contribute to intimal formation in atherosclerosis, after arterial injury and in transplant atherosclerosis. The precise frequency of these bone marrow-derived vascular precursor cells is controversial and their role is unknown. In addition, additional data support the presence of a resident progenitor cell subpopulation and its involvement in the response of the adult arterial wall to damage or ischemia. This review will examine the evidence for and the putative role of progenitor cell-derived VSMCs in arterial disease, a necessary prerequisite before deciding whether progenitor cells are therapeutic targets in vascular disease.     

富含微量元素矿泉水对人血管平滑肌细胞生长增殖的影响

目的研究富含微量元素矿泉水对体外培养血管平滑肌细胞（HVSMCs）生长增殖的影响。方法无菌取12例冠状动脉旁路移植术患者术中废弃的大隐静脉,应用金山矿泉水配制的DMEM培养液（MW—DMEM）和双蒸水配制的DMEM培养液（DDW—DMEM）分别培养平滑肌细胞,分为金川矿泉水组和双蒸水组。观察记录2种培养液培养的HVSMCs游出组织块的时间及细胞呈“峰”-“谷”样时间;建立血管紧张素Ⅱ诱导HVSMCs增殖模型,采用MTT比色法观察2种培养液对AngⅡ诱导HVSMCs增殖抑制作用;应用电镜和免疫组织化学染色法观察鉴定HVSMCs。结果金川矿泉水组培养的HVSMCs贴壁时间为（3．7±0．2）d,双蒸水组为（8．5±0．6）d;金川矿泉水组培养的HVSMCs自贴壁后到长成“峰”-“谷”样形态的时间为（8．3±0．2）d,双蒸水组为（7．1±0．5）d;MTT比色法测定金川矿泉水组培养的HVSMCs的增殖活度明显降低;电镜观察MW—DMEM培养的HVSMCs细胞形态完整,表面有丰富的微绒毛。HVSMCs胞浆内富含肌丝,纵向平行排列,胞内有密体;抗-actin平滑肌肌动蛋白免疫组织化学染色后可见胞浆内染成褐色呈细丝网状。结论富含微量元素和矿物质的金川矿泉水具有修复血管平滑肌细胞损伤和抑制血管紧张素Ⅱ诱导HVSMCs增殖的作用。     

血管内皮生长因子受体-3在血管内皮细胞和平滑肌细胞中的表达

目的探讨血管内皮细胞生长因子受体-3（VEGFR-3）在血管内皮细胞和平滑肌细胞中的表达。方法分离人脐带中的脐静脉和脐动脉,冰冻切片后以免疫荧光法检测VEGFR-3的表达。结果VEGFR-3荧光信号表达于脐静脉内皮细胞和平滑肌细胞,以及脐动脉内皮细胞、平滑肌细胞和外周细胞。结论血管内皮细胞和平滑肌细胞均表达VEGFR-3。     

The effect of forskolin on smooth muscle cells of guinea-pig taenia caeci

Forskolin (0.5-10 microM) caused hyperpolarization and relaxation of the smooth muscle cells of guinea-pig taenia caeci (35 degrees C) as measured with the sucrose-gap method. Membrane conductance, reflected by the amplitude of the electrotonic potential, was not changed during the response. The hyperpolarization could also be evoked in the absence of extracellular calcium, calcium and potassium, or calcium and sodium. Further, the 45Ca2+ efflux from taenia caeci was enhanced by forskolin. The results support the concept that calcium extrusion across the plasma membrane is promoted in the presence of forskolin by stimulation of electrogenic calcium pumping.     

Threshold of peroxynitrite cytotoxicity in bovine pulmonary artery endothelial and smooth muscle cells

Peroxynitrite is widely reported as highly cytotoxic; yet recent evidence indicates that at certain concentrations, it can induce pulmonary cell hyper-proliferation and tissue remodelling. This study aimed to establish the threshold concentration of peroxynitrite to induce functional impairment of bovine pulmonary artery endothelial (PAEC) and smooth muscle cells (PASMC). PAEC or PASMC were exposed to solution of peroxynitrite or 3-morpholinosydnonimine (SIN-1). Twenty-four hour cell viability, DNA synthesis, and protein biochemistry were assessed by trypan blue dye exclusion, [³H] thymidine incorporation and western blot analysis, respectively. Threshold concentration of peroxynitrite to significantly impair viability of PAEC and PASMC was 2 μM peroxynitrite. In PASMC and PAEC, low concentrations of peroxynitrite (2 nM–0.2 μM) increased cell proliferation and did not activate p38 MAP kinase. The decrease in DNA synthesis and cell viability caused by 2 μM peroxynitrite was associated with caspase-3 cleavage but not p38 activation. Also, 2–20 μM peroxynitrite significantly activated poly ADP ribose polymerase and stress activated kinase JNK in PAEC. However, the higher concentration of 20 μM peroxynitrite did cause a threefold increase in p38 activation. In conclusion, the threshold for the cytotoxic effects of peroxynitrite was 2 μM; which caused apoptotic cell death independent of p38 MAP kinase activation in pulmonary artery cells.     

同型半胱氨酸诱导人血管平滑肌细胞氧化损伤的研究

目的探讨同型半胱氨酸（Hcy）对人脐静脉血管平滑肌细胞（VSMCs）氧化损伤的作用及机制。方法用含不同剂量Hcy（50～1000μmol/L）的培养基培养VSMCs24h后,以分光光度比色法分别测定细胞内丙二醛（MDA）含量和超氧化物酶（SOD）、过氧化氢酶（CAT）活力的改变。结果随Hcy浓度增加,VSMCs内MDA的含量明显增加,SOD和CAT的活力也显著升高。Hcy明显促进了VSMCs的氧化。结论Hcy可直接诱导VSMCs的氧化损伤。     

Angiotensin II-induced responses in vascular smooth muscle cells: inhibition by non-peptide receptor antagonists

The present study investigates the effect of angiotensin II and LR-B/081 (-methyl 2-[[4-butyl-2-methyl-6-oxo-5-[[2′-(1H-tetra-zol-5-yl) [1,1′-biphenyl]-4-yl] methyl]-1(6H)-pyrimidinyl] methyl]-3-thiophenecarboxylate), a novel non-peptide angiotensin II receptor antagonist, on both early and late responses in rat vascular smooth muscle cells. Angiotensin II induced a rapid and transient elevation of inositol trisphosphate intracellular levels, triggered the release of both prostaglandin E₂ and prostaglandin I₂ (EC₅₀ = 21 ± 3 and 16 ± 2 nM, respectively), and, in long-term studies, increased leucine and thymidine incorporation. All angiotensin II effects were antagonized by LR-B/081 and losartan, the reference non-peptide angiotensin AT₁-selective receptor antagonist, whereas they were unaffected by PD123177 (1-(4-amino-3-methylphenyl)methyl-5-diphenylacetyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine carboxylic acid), a non-peptide angiotensin AT₂-selective receptor antagonist. LR-B/081 displayed a much higher potency than losartan in inhibiting angiotensin II-induced prostaglandin E₂ (IC₅₀ = 0.15 ± 0.02 and 39 ± 9 nM, respectively) and prostaglandin I₂ release (IC₅₀ = 0.18 ± 0.04 and 134 ± 40 nM, respectively) and was also more potent in blocking the increase in protein synthesis (IC₅₀ = 242 ± 119 nM and 1221 ± 687 nM, respectively). Moreover, LR-B/081 and losartan blocked the response to angiotensin III but failed to inhibit the prostaglandin release stimulated by vasopressin or the mitogenic effect of serum. LR-B/081 and losartan were devoid of intrinsinc agonistic properties in the experimental conditions employed. The present results describe LR-B/081 as a novel, highly specific and potent, non-peptide angiotensin AT₁-selective receptor antagonist, that is capable of blocking angiotensin II-proliferative responses, which may be of relevance for cardiovascular diseases.     

血管平滑肌细胞表型转化的分子机制研究进展

血管内膜新生是血管成形术后再狭窄和动脉粥样硬化等心血管疾病的病理基础。血管平滑肌细胞(vascular smooth muscle cells, VSMCs)具有高度可塑性,在众多病理因素下常发生表型转化(phenotypic switch),即由收缩型向合成型转化。研究表明,血管平滑肌细胞表型转化促进其增殖和迁移,从而参与血管内膜新生形成。本文主要综述了血管平滑肌细胞表型转化影响内膜新生的分子机制,为防治血管重塑性疾病提供理论依据。     

糖尿病大鼠冠状动脉平滑肌细胞体外培养方法的探讨

目的探讨糖尿病大鼠冠状动脉平滑肌细胞体外培养方法,建立糖尿病大鼠冠状动脉平滑肌细胞模型,为糖尿病性冠心病的研究奠定基础。方法建立糖尿病大鼠模型,用酶消化法体外培养冠状动脉血管平滑肌细胞。结果糖尿病大鼠造模成功后分离冠状动脉,用酶消化法培养糖尿病大鼠血管平滑肌细胞,24h更换培养液液,培养7~10d细胞重叠生长达多层,高低起伏呈“峰—谷”状。细胞α-actin免疫组织化学染色鉴定为平滑肌细胞。结论糖尿病大鼠血管平滑肌细胞增殖速度快,培养条件要求严格,在形态学上与正常大鼠平滑肌细胞相同。