Wnt信号通路在血管钙化中作用的研究进展

血管钙化是一种主动的可调控的类似于骨或软骨性发育的复杂过程,涉及许多疾病的发病过程,如动脉粥样硬化、高血压、糖尿病及慢性肾病等。研究发现Wnt信号通路以旁分泌或自分泌的方式参与心脏发育、心肌肥厚、动脉粥样硬化及血管再生等病理生理过程。近年来,Wnt信号通路在血管钙化发病过程中的作用亦日益受到重视。     

慢性肾脏病血管钙化发病机制的研究进展

血管钙化(vascular calcification,VC)是慢性肾脏病(chronic kidney disease,CKD)患者心血管病死率高的主要原因.VC发病机制非常复杂,与肾的排泄功能、内分泌功能减退等相关,如明显的钙磷代谢紊乱、klotho蛋白水平降低、炎症氧化应激和细胞凋亡、钙化抑制因子的减少和尿毒症毒...     

维生素D在血管钙化中的研究进展

本文综述维生素D预防和治疗血管钙化方面的研究进展。主要综述了国内外的相关文献对维生素D的抗血管钙化作用和最新研究,基础实验和临床观察都显示维生素D对血管钙化具有重要的调控作用,目前主要研究热点是维生素D抑制血管平滑肌细胞(vascular smooth muscle cell,VSMC)的表型转化,主要问题是维生素D在血管钙化中对血管平滑肌细胞表型的调控作用,且其发挥血管保护作用的机制尚需更多的研究验证。     

内皮素受体阻断剂抑制磷酸甘油诱导血管平滑肌细胞钙化

目的：在离体钙化的血管平滑肌细胞上，观察内皮素受体阻断剂对血管平滑肌细胞(VSMCs)钙化的影响，探讨内皮素(ET)促进细胞钙化的信号转导和分子机制。方法：β-磷酸甘油制备钙化VSMCs；通过细胞钙含量,［45Ca2+］摄取，碱性磷酸酶活性测定，判断钙化程度，［3H］-胸腺嘧啶（［3H］-TdR）和［3H］-亮氨酸（［3H］-Leu）掺入测定细胞DNA合成，竞争性定量RT-PCR测定VSMCs骨桥蛋白 （OPN） mRNA水平，放射免疫法测定培养上清ET含量。结果: 与正常对照VSMCs相比，钙化VSMCs内钙含量，［45Ca2+］摄入及碱性磷酸酶活性分别增加 119%、174%和7倍（P<0.01），OPN表达增加86%；细胞生长活跃，［3H］-TdR和［3H］-Leu分别增加71%和35%；钙化细胞培养基中内皮素含量较对照组增加35%(P<0. 05)。而钙化加内皮素受体阻断剂BQ123组明显减轻VSMCs钙化程度，钙含量，［45Ca2+］摄入及碱性磷酸酶活性分别较钙化组降低33%、37%、40%（均P<0.01），OPN表达明显下调25%；细胞增殖活性明显降低。结论: BQ123可减轻VSMCs钙化程度，表明ET促进VSMCs钙化主要是通过内皮素A型受体（ET-A）途经。     

内皮素受体阻断剂抑制磷酸甘油诱导血管平滑肌细胞钙化木

目的:在离体钙化的血管平滑肌细胞上,观察内皮素受体阻断剂对血管平滑肌细胞(VSMCs)钙化的影响,探讨内皮素(ET)促进细胞钙化的信号转导和分子机制.方法:磷酸甘油制备钙化VSMCs;通过细胞钙含量,[45Ca2 ]摄取,碱性磷酸酶活性测定,判断钙化程度,[3H]-胸腺嘧啶([3H]-TdR)和[3H]-亮氨酸([3H]-Leu)掺人测定细胞DNA合成,竞争性定量RT-PCR测定VSMCs骨桥蛋白(OPN)mRNA水平,放射免疫法测定培养上清ET含量.结果:与正常对照VSMCs相比,钙化VSMCs内钙含量,[45ca2 ]摄入及碱性磷酸酶活性分别增加119％、174％和7倍(P<0.01),OPN表达增加86％;细胞生长活跃,[3H]-TdR和[3H]-Leu分别增加7l％和35％;钙化细胞培养基中内皮素含量较对照组增加35％(P<0.05).而钙化加内皮素受体阻断剂BQl23组明显减轻VSMCs钙化程度,钙含量,[45ca2 ]摄入及碱性磷酸酶活性分别较钙化组降低33％、37％、40％(均P<0.01).OPN表达明显下调25％;细胞增殖活性明显降低.结论:BQl23可减轻VSMCs钙化程度,表明ET促进VsMcs钙化主要是通过内皮素A型受体(ET-A)途经.     

淫羊藿苷对大鼠血管平滑肌细胞钙化的抑制作用

目的:研究淫羊藿苷(Icariin)对大鼠血管平滑肌细胞(VSMCs)钙化的抑制作用。方法:组织贴壁法培养大鼠胸主动脉VSMCs,取5-8代细胞分为对照组、钙化模型组和淫羊藿苷干预A、B、C组。对照组用DMEM培养基培养;钙化模型组用含10mmol/Lβ-甘油磷酸盐(β-GP)的DMEM培养基培养;干预A、B、C组均以钙化模型组为基础分别与10-6 mol/L、10-5 mol/L和10-4 mol/L Icariin的DMEM培养基共培养,茜素红-S染色法鉴定各组细胞钙化情况,硝基苯酚法测定各组细胞碱性磷酸酶(ALP)活性,RT-PCR检测各组细胞内骨保护素(OPG)和核因子κB(NF-κB)受体活化因子配体(RANKL)的mRNA水平,用Western blotting检测各组细胞OPG和RANKL的蛋白表达。结果:与对照组比较,钙化模型组VSMCs发生细胞钙化,并形成红色结节,ALP活性显著升高(P0.01),OPG、RANKL mRNA和蛋白表达均增加(P0.01);与钙化模型组比较,干预A、B、C组细胞钙化减少,ALP活性减低(P0.01),OPG、RANKL mRNA和蛋白表达均下调(P0.01),尤以干预B组效果最显著。结论:淫羊藿苷可能通过降低ALP活性及OPG、RANKL表达来抑制动脉钙化。     

大鼠钙化血管平滑肌细胞血红素加氧酶-CO-cGMP通路的变化

本文旨在观察钙化细胞血红素加氧酶 (HO) -一氧化碳 (CO) -环磷酸鸟苷 (cGMP)系统的改变 ,以探讨血管钙化发生的细胞分子机理。利用 β 甘油磷酸制备钙化血管平滑肌细胞 (VSMCs) ,测定细胞钙含量、碱性磷酸酶活性及4 5Ca沉积 ;观察HO 1免疫细胞化学染色 ;测定VSMCsHO 1活性、碳氧血红蛋白 (HbCO)的生成及cGMP含量。发现钙化细胞的钙含量、碱性磷酸酶活性及4 5Ca沉积均比正常组显著升高。钙化细胞HO 1的表达不均匀 ,钙化结节中HO 1表达强于正常细胞。与正常平滑肌细胞相比 ,钙化细胞的HO 1活性低 4 2 7% [36 4± 2 8pmol (mgprotein·h)vs 6 3 5± 5 3pmol (mgprotein·h) ,P <0 0 1];CO含量低 39 2 % (3 38± 0 39μmol mgproteinvs 5 5 6± 0 4 8μmol mgprotein ,P <0 0 5 ) ;cGMP含量比正常细胞低 78 1% (4 3± 0 5vs 19 6± 1 2pmol mgprotein ,P <0 0 1)。提示钙化血管血红素 HO CO cGMP途径功能下调     

体表血管平滑肌瘤及其钙化亚型的临床病理学观察

目的 探讨体表性血管平滑肌瘤(cutaneous angioleiomyoma,CAL)及其钙化亚型(cutaneous calcified angioleiomyoma,CCAL)的临床病理学特征.方法 对61例CAL(包括4例CCAL)的临床表现、影像学特征及病理学特点进行分析.结果 女性多见,占60.66%(37/61),就诊年龄10～87岁,平均43.08岁,以20～69岁居多,占85.25%(52/61).4例CCAL均为女性,平均年龄63.75岁.所有病例中以下肢受累最为多见,占75.41%(46/61).临床表现为局部真皮深层或皮下缓慢性生长的多于半数伴有疼痛(35/61,57.38%)的孤立性结节,局部皮肤稍隆起,偶尔呈息肉样外观.肿瘤无特征性影像学表现,其显示病变为真皮深层或皮下孤立性结节,少数病变可伴有程度不等的钙化.大体示肿瘤结节质硬,界清,有纤薄的假包膜,直径为0.5～3.5 cm,圆形或卵圆形.镜下肿瘤实质由丰富的血管和增生的平滑肌混合而成.根据主要组织学结构分为3型:实性型、静脉型和海绵型.少数病变可伴发玻璃样变、钙化、黏液样变、脂肪组织增生、淋巴细胞浸润、出血及脉管内血栓机化等改变.CCAL病变中钙化灶呈沙粒状、斑块状或无定型团块状,钙化显著者可占据肿瘤实质的绝大部分区域,仅残留少量难于识别的平滑肌组织.免疫组化检查显示病变组织中平滑肌细胞表达vimentin、desmin、MSA、α-SMA、calponin及caldemon,而HMB45及CD34均阴性.结论 CAL是一种发生于体表的、可有疼痛的良性软组织肿瘤,位于真皮深层或皮下组织内,影像学无特征性改变,形态上主要表现为增生的平滑肌束围绕在血管周围呈肿瘤性的生长.少数病例可伴发程度不等的钙化,诊断时需与肿瘤性钙盐沉积症、钙化上皮瘤、钙化性纤维性假瘤以及其他继发性皮肤钙盐沉积症相鉴别.     

细胞在小口径组织工程血管中的抗钙化作用

背景：小口径组织工程血管的远期结果研究极少,尚未见到研究组织工程血管分子水平、离子水平远期结果和平滑肌细胞与钙化关系的报道。 目的：利用脱细胞猪股动脉基质作为支架和犬血管壁细胞作为种子细胞体外构建小口径组织工程血管,植入种子细胞供体犬股动脉部位6个月,观察植入物中层平滑肌细胞与钙化的关系。 方法：12只实验犬被随机分为支架组(n=6)和再细胞化组(n=6),自体股动脉被作为对照组;支架组犬接受猪股动脉经脱细胞后的基质支架植入双侧股动脉,再细胞化组犬接受受体血管壁细胞共同培养、联合种植于脱细胞的猪股动脉基质并体外预适应后植入血管壁细胞供体双侧股动脉位置;6个月后测定植入物和对照组股动脉组织钙含量、平滑肌密度和病理学变化。 结果与结论：小口径组织工程血管植入后6个月检查见2组植入物无明显狭窄和扩张,扫描电镜示内表面均已完全内皮化,有管壁僵硬和局部钙化斑块形成,以上改变以支架组植入物更明显。支架组管道组织钙含量显著高于再细胞化组和自体股动脉(P < 0.01),再细胞化组植入物组织钙含量亦显著高于自体股动脉(P < 0.01);病理学检查示再细胞化组植入物平滑肌密度高于支架组(P < 0.01),再细胞化组和支架组植入物平滑肌密度均低于对照组(P < 0.01);超声检查见2组管道植入术后即刻与6个月后舒缩幅度较邻近自体股动脉舒缩幅度小,有部分管道无舒缩功能。结果提示,猪股动脉常规脱细胞方去获得的基质作为支架体外构建组织工程血管时,平滑肌细胞难于迁移至支架中层,中层平滑肌密度低,植入体内6个月后中层平滑肌细胞密度仍低,平滑肌细胞有抗血管钙化作用。     

miR-30b在高磷诱导的大鼠血管平滑肌细胞钙化中的作用

目的:观察微小RNA-30b(miR-30b)在高磷诱导的大鼠血管平滑肌细胞(VSMCs)钙化中的作用及其机制.方法:将SD大鼠随机分为假手术(sham)组和慢性肾衰竭所致血管钙化(CRF+VC)组.采用硝酸银染色检测血管钙化情况;免疫组织化学法检测成骨转录因子Runx2的表达.体外培养原代大鼠胸主动脉VSMCs并分为...     

CDC42 promotes vascular calcification in chronic kidney disease

Vascular calcification is prevalent in patients with chronic kidney disease (CKD) and a major risk factor of cardiovascular disease. Vascular calcification is now recognised as a biological process similar to bone formation involving osteogenic differentiation of vascular smooth muscle cells (VSMCs). Cell division cycle 42 (CDC42), a Rac1 family member GTPase, is essential for cartilage development during endochondral bone formation. However, whether CDC42 affects osteogenic differentiation of VSMCs and vascular calcification remains unknown. In the present study, we observed a significant increase in the expression of CDC42 both in rat VSMCs and in calcified arteries during vascular calcification. Alizarin red staining and calcium content assay revealed that adenovirus-mediated CDC42 overexpression led to an apparent VSMC calcification in the presence of calcifying medium, accompanied with up-regulation of bone-related molecules including RUNX2 and BMP2. By contrast, inhibition of CDC42 by ML141 significantly blocked calcification of VSMCs in vitro and aortic rings ex vivo. Moreover, ML141 markedly attenuated vascular calcification in rats with CKD. Furthermore, pharmacological inhibition of AKT signal was shown to block CDC42-induced VSMC calcification. These findings demonstrate for the first time that CDC42 contributes to vascular calcification through a mechanism involving AKT signalling; this uncovered a new function of CDC42 in regulating vascular calcification. This may provide a potential therapeutic target for the treatment of vascular calcification in the context of CKD. © 2019 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

血管平滑肌细胞凋亡在血管再狭窄中作用的研究进展

血管平滑肌细胞构成新生内膜增生的重要部分,且在血管腔内治疗术后再狭窄的心血管疾病的发生和发展中具有重要作用。血管平滑肌细胞凋亡能有效抑制血管球囊损伤和血管旁路移植术后新生内膜增生,从而可为血管术后再狭窄提供治疗手段。     

Electromagnetic fields promote severe and unique vascular calcification in an animal model of ectopic calcification

BackgroundThe effects of electromagnetic fields (EMFs) on cardiovascular calcification is unknown. We sought to evaluate the effects of EMF on vascular calcification in normal rats and in rats with chronic kidney disease (CKD) – a condition which promotes calcification.MethodsWe used four groups of rats: group 1 – exposed to EMF, group 2 – not exposed to EMF, group 3 – rats with CKD exposed to EMF, group 4 – rats with CKD not exposed to EMF. In order to induce CKD, groups 3 and 4 rats were fed with a uremia-inducing diet. Groups 1 and 3 rats were continuously exposed to EMF using a system similar to an electrical transformer, which consists of a primary coil, a ferrite ring, and a secondary coil. The system transmitter emitted a series of exponentially decaying electromagnetic sine waves (continuous exposure with pulsed peaks) in randomly selected frequencies between 150 and 155 kHz, with random exposure intensities between 4 and 7 mG. Clinical investigations included multislice computed tomography of the aortic roots. Pathological examinations of the aortas included histological characterization, and antigen expression analyses.ResultsNo calcification was found in either group of rats with normal kidney function. Aortic root calcification was significantly higher in rats exposed to EMF (group 3) compared with group 4 rats – with a mean Agatston score of 138±25 vs. 80±20 respectively (p < 0.05). Pathological examination showed massive aortic calcification in group 3 rats. The calcification pattern was unique as it formed circular rings along the length of the aortic media.Although increased calcification was noticed in group 3 rats, antigen expression of osteoblast markers was significantly decreased in group 3 compared with group 4.ConclusionsEMF exposure may have potential harmful effects on the cardiovascular system, as it promotes severe vascular calcification in CKD miliue.     

雌激素受体在大鼠血管平滑肌细胞中的表达及其调控

目的：观察血管平滑肌细胞(VSMC)中雌激素受体(ER)的表达,研究雌激素和血清(含多种生长因子)对细胞中：ER表达的影响。方法：原位杂交和RT-PCR检测大鼠主动脉中层VSMC中的ERmRNA。分别在有或无他莫昔芬(TAM)(ER拮抗剂)预处理的情况下,免疫细胞化学观察VSMC中的ER蛋白,比较雌二醇(E2)以及新生小牛血清(NCS)对VSMC中ER表达的影响。结果：VSMC中存在ERmRNA和蛋白,E2以及NCS均能上调细胞中ER的表达,TAM能阻断其效应。结论：ER是雌激素作用于VSMC的靶基因之一。     

Citrate attenuates vascular calcification in chronic renal failure rats

Vascular calcification (VC) is a major contributor of cardiovascular dysfunction in chronic renal failure (CRF). Citrate binds calcium and inhibits the growth of calcium crystals. This present study intends to evaluate the effect of citrate on VC in adenine‐induced CRF rats. The rats were randomly divided into five groups: the control group, the citrate control group, model group, model rats with low‐dose treatment of citrate (216 mg/kg) and model rats with high‐dose treatment of citrate (746 mg/kg). The rats were euthanized at 5 weeks with their blood and aorta in detection. The results showed that serum level of blood urea nitrogen, serum creatinine, phosphorus, calcium, and related renal failure function marker were elevated in the model group. Furthermore, the aortic calcium accumulation and alkaline phosphatase activity were significantly increased in the model group compared with control groups. Additionally, hematoxylin–eosin staining results demonstrated that the vascular calcification in aorta is significantly increased in the model group. Finally, the expression of VC‐related proteins including bone morphogenetic protein and osteocalcin were increased in the model group, whereas alpha‐smooth muscle actin was decreased in the model group compared with the control group. However, treatment with citrate caused a reversal effect of all the above events in a dose‐dependent manner. In conclusion, citrate may attenuate vascular calcification in adenine‐induced CRF rats.     

牛主动脉平滑肌细胞体外钙型的制备

目的利用β-甘油磷酸盐处理牛主动脉平滑肌细胞(BASMC)制备体外血管钙化模型。方法移植块法原代培养牛主动脉中膜平滑肌细胞,8代内的传代细胞添加10 mmol/Lβ-甘油磷酸盐培养10 d,Von Kossa染色、茜素红S染色和电镜检查鉴定钙化,同时测量细胞层钙沉积、碱性磷酸酶活性及培养上清的骨钙素含量。结果10 d后细胞层出现大量钙盐沉积,并且β-甘油磷酸盐时间依赖性增加钙沉积,碱性磷酸酶活性及骨钙素含量各时间点均较正常培养细胞显著增加(P<0.01)。结论甘油磷酸盐能在短期内诱导出BASMC广泛钙化,用此方法制备的BASMC是一种良好的研究血管钙化的体外模型。