全反式维甲酸抑制血管平滑肌增殖的作用机制

血管平滑肌细胞（vascular smooth muscle cells,VSMC）的增殖是动脉粥样硬化等血管增生性疾病的重要病理特征。因此,抑制VSMC增殖及促进其分化的药物均有望用于动脉粥样硬化的治疗。全反式维甲酸（ATRA）是具有广泛生物学效应的维生素A类物质。研究证实ATRA可与VSMC中存在的维甲酸类受体结合,调控VSMC从低分化的合成型转变为成熟的收缩袁型,进而防止其过度增殖。     

全反式维甲酸体外诱导P19细胞向心肌方向分化

目的探讨不同浓度全反式维甲酸(all-trans retinoic acid,atRA)诱导P19细胞向心肌分化的效力。方法细胞分成P19细胞组,2nm/L atRA诱导组,5nm/L atRA诱导组,8nm/L atRA诱导组。各组细胞经过诱导、聚集培养、聚集体贴壁培养10天后,RT-PCR检测GATA-4,α-肌球蛋白重链(α-myosin heavychain,α-MHC)的mRNA表达,免疫荧光双标检测α-sarcomeric actin和cTnT蛋白共表达,Western blot检测cTnT的蛋白表达。结果 atRA可诱导聚集P19细胞表达GA-TA-4、a-MHC mRNA;α-sarcomeric actin和cTnT的表达和共表达增加;5nm/L atRA组,8nm/L atRA组GATA-4、a-MHCmRNA的表达量显著高于P19细胞组;5nm/L atRA组,8nm/L atRA组两种蛋白的表达和共表达量显著高于P19细胞组,以5nm/L atRA组最高,显著高于其它组。结论 atRA可诱导聚集P19细胞向心肌分化,其中5nm/L atRA组效果最好。     

藻蓝蛋白联合全反式维甲酸对HeLa细胞生长抑制作用的研究(英文)

目的:探讨全反式维甲酸和藻蓝蛋白单独及联合用药对HeLa细胞生长的影响,并揭示两者联合用药对细胞周期和细胞凋亡影响的分子机制。方法:MTT法检测全反式维甲酸和藻蓝蛋白单独及联合用药对HeLa细胞生长的影响,原位杂交法检测用药前后细胞内CDK-4基因mRNA的表达情况,免疫组化法检测用药前后bcl-2基因的表达情况,TUNEL法检测用药前后细胞凋亡情况。结果:全反式维甲酸和藻蓝蛋白均具有抑制HeLa细胞生长的作用,当达到相同的抑制率时,联合藻蓝蛋白使用可以显著降低全反式维甲酸的使用剂量从而达到降低毒副作用的目的。两者联合用药可以显著降低CDK-4的表达量从而对HeLa细胞的细胞周期产生影响。两者联合用药可以显著下调bcl-2的表达水平从而引发细胞凋亡。结论:通过联合藻蓝蛋白,可以显著降低全反式维甲酸的使用剂量从而降低毒副作用。全反式维甲酸和藻蓝蛋白联合用药抑制HeLa细胞生长的分子机制可能是通过抑制CDK-4和bcl-2的表达来影响细胞周期并最终导致细胞凋亡。     

全反式维甲酸诱导胃腺癌细胞周期阻滞的作用机制

目的:研究全反式维甲酸(all trans retinoic acid,ATRA)对人胃腺癌细胞(SGC-7901)细胞周期阻滞的诱导作用并探讨其机制.方法:采用四甲基偶氮唑盐(MTT)方法检测ATRA对SGC-7901增殖的抑制;流式细胞术检测细胞周期,Western blot方法检测不同浓度ATRA处理后的SGC-7901细胞中Akt、P-Akt(Ser473)、P-Akt(Thr308)、p-GSK-3β(Ser9)和cyclin D1的表达情况.结果:10.9～10.5mol/L的ATRA作用SGC-7901细胞48 h,能显著抑制细胞增殖,其抑制率分别为10.2%±0.5%、15.3%±0.5%、17.0%±0.7%、28.4%±1.0%和36.9%±0.7%;G1期细胞比率随着ATRA浓度的增加而增加,呈明显的G1期阻滞;Western blot检测显示ATRA对细胞中Akt蛋白的表达没有明显影响,两种p-Akt蛋白的表达显著下调,ATRA显著降低细胞中cyclin D1和p-GSK-3β的表达.结论:ATRA可能通过抑制磷酸化Akt蛋白表达而减少p-GSK-3β的表达,从而减少cyclin D1的表达量,进而诱导SGC-7901细胞发生G1期阻滞.     

血管平滑肌细胞表型调节机制的研究进展

血管平滑肌细胞（VSMC）的增殖和迁移是动脉粥样硬化斑块形成、高血压和血管再狭窄的共同病理特征,而VSMC表型转化是VSMC增殖和迁移的基础,研究VSMC表型调节的分子机制,对上述疾病的防治具有重要意义。本文对VSMC表型转化的影响因素、信号转导途径和转录因子的研究进展作一综述。     

血管平滑肌细胞在动脉粥样硬化中的作用

动脉粥样硬化的发生发展是一个复杂的过程,涉及到多种细胞及细胞因子的相互作用.平滑肌细胞作为血管壁的重要成分,调节着血管的收缩舒张功能,同时也分泌多种细胞因子及细胞间质;它的生物学行为对动脉粥样硬化的发生、发展及最终的结局产生着重要的影响.本文就平滑肌细胞的生物学行为的变化及其在动脉粥样硬化的不同发展阶段的作用进行综述.     

平滑肌22α蛋白在血管稳态和血管重构中的作用

有关血管稳态和重构的分子机制一直是近年来的研究热点,也被视为治疗血管损伤性疾病的突破点.大量研究证实,血管损伤修复及病理性重构过程与血管平滑肌细胞(vascular smooth muscle cells,VSMCs)的表型转化、异常增殖与迁移、细胞衰老关系密切.平滑肌22α(smooth muscle 22α,SM2...     

全反式维甲酸对哮喘模型大鼠Th1/Th2平衡的影响

[目的]观察不同剂量全反式维甲酸(All-trans retinoid acid, ATRA)对哮喘模型大鼠Th1/Th2平衡的影响。[方法]将SD大鼠随机分为对照组、模型组、低剂量组、中剂量组和高剂量组。对照组和模型组的大鼠给予生理盐水(10 mL/kg)灌胃,对低/中/高剂量组的大鼠,分别给予10 mg/kg、20 mg/kg和40 mg/kg ATRA灌胃,1次/d,持续1 w。观察各组大鼠的肺功能、肺组织匀浆液细胞因子水平、血Th1/Th2值及Notch通路的表达情况。[结果]中/高剂量组大鼠呼吸频率显著低于模型组(P<0.05);吸气流量、呼气峰流量和潮气量显著高于模型组(P<0.05);模型组、低/中/高剂量组肺组织匀浆液中的IgE、IL-4和IL-5水平依次降低,IFN-γ和IL-12水平依次升高;模型组Th1/Th2值显著高于对照组(P<0.05),ATRA治疗后Th1/Th2值较模型组均显著降低,剂量越高Th1/Th2值降低越明显;与对照组相比,模型组、低/中/高剂量组的Notch3、Delta1、Jagged1和Jagged2的mRNA和蛋白表达水...     

影响主动脉平滑肌细胞迁移的因素

平滑肌细胞(vascular smooth muscle cell,VSMC)的迁移对血管发育、动脉粥样硬化和术后再狭窄等起到关键性的作用。主要从激发VSMC迁移的关键炎性细胞因子、细胞间相互作用的核心成员、microRNA、细胞骨架和上述各因素的迁移信号通路这几方面来综述VSMC的迁移。     

Intracellular superoxide induces apoptosis in VSMCs: role of mitochondrial membrane potential,cytochrome C and caspases

Apoptosis of vascular smooth muscle cells (VSMCs) is an integral part of cardiovascular diseases including atherosclerosis, hypertension and restenosis. Here we studied the fate of VSMCs in response to intracellular superoxide stimulation. Diethyldithiocarbamic acid (DDC) was used to inhibit copper-zinc superoxide dismutase thereby increasing intracellular superoxide levels. The results show that DDC at a dose from 25–100 M is able to induce VSMC apoptosis. Superoxide was found to be responsible for DDC-induced apoptosis. In the apoptotic process mitochondrial membrane potential was decreased and caspase-3, -8 and -9 were activated. Surprisingly, neither cytochrome c release nor Bid cleavage could be observed. These data suggest a role for intracellular superoxide in the regulation of VSMCs apoptosis.     

全反式维甲酸通过抑制ERK和激活p38 MAPK信号诱导KLF4基因表达

全反式维甲酸（all-trans retinoic acid, ATRA）诱导细胞分化与上调转录因子Krüppel样因子4 (KLF4)表达有关, 但目前对ATRA诱导KLF4表达的分子机制尚不清楚．为了研究ATRA在血管平滑肌细胞（VSMC）中诱导KLF4表达的分子机制,本 研究观察ATRA对视黄酸受体α (retinoic acid receptor α, RARα)和KLF4表达的影响及RARα介导ATRA诱导KLF4表达所依 赖的信号转导途径．实验结果显示,ATRA可显著诱导RARα和KLF4表达,用RARα拮抗剂Ro 41 5253阻断ATRA与受体相互作 用后,ATRA诱导的KLF4表达受到显著抑制．用p38 MAPK、ERK和Akt抑制剂阻断ATRA与RARα相互作用所激活的信号转导途径 后,发现阻断p38 MAPK信号途径显著抑制ATRA诱导的KLF4表达,抑制ERK信号途径使ATRA对KLF4表达的诱导作用明显增强, 抑制Akt信号途径不影响KLF4基因表达．表明RARα介导ATRA对KLF4表达的诱导作用,ATRA通过抑制ERK和激活p38 MAPK信号 途径发挥其对KLF4基因表达的诱导作用．     

Macrophage migration inhibitory factor (MIF) promotes rat airway muscle cell proliferation and migration mediated by ERK1/2 and FAK signaling

Macrophage migration inhibitory factor (MIF) is an inflammatory mediator that contributes to asthmatic airway remodeling; however, little is known regarding the effects of MIF on airway smooth muscle cells (ASMCs). In the present study, we found that an enhanced expression of MIF promoted ASMC proliferation, increased the population of cells in the S/G2 phase, downregulated P21 expression, and upregulated cyclin D1, cyclin D3, and Cdk6 expression. In addition, the apoptosis of ASMCs was significantly decreased in response to MIF overexpression, compared with the negative control. Moreover, MIF facilitated the migration of ASMCs by upregulating the expression of matrix metalloproteinase (MMP)‐2. Finally, we showed that MIF increased the phosphorylation of extracellular regulated protein kinases (ERK) 1/2 and focal adhesion kinase (FAK), which are associated with proliferation and migration. In conclusion, this study demonstrated that MIF overexpression promotes the proliferation and migration of ASMCs by upregulating the activity of the ERK1/2 and FAK signaling pathways in these cells, further indicating that inhibition of MIF may prove to be an effective strategy for treating asthma patients with airway remodeling.     

Preliminary study on the mechanism of long noncoding RNA SENCR regulating the proliferation and migration of vascular smooth muscle cells

The proliferation and migration of vascular smooth muscle cells (VSMCs) are one of the key regulatory links of atherosclerosis (AS). Long noncoding RNAs (lncRNAs) are emerging as key regulators in AS development. In this study, we first assessed the expression level of smooth muscle and endothelial cell-enriched migration/differentiation-associated lncRNA (SENCR) in the plasma of patients with coronary heart disease (CHD) and its predictive and diagnostic value. Second, we investigated the role of SENCR in the regulation network of human aortic-VSMCs (HA-VSMCs) proliferation and migration and determined its downstream regulatory mechanism. The results showed that SENCR was downregulated in the peripheral blood of CHD, and negatively related to the Gensini score. SENCR was enriched in HA-VSMCs and mainly distributed in cytoplasm. Overexpression of SENCR significantly inhibited HA-VSMCs proliferation, migration, and block cell cycle, while the knockdown of SENCR had the opposite effects. Moreover, bioinformatics analysis and luciferase reporter assay demonstrated that miR-4731-5p could directly bind to SENCR. Besides, we proved that FOXO3a inhibited HA-VSMCs proliferation and migration by binding to the 3′-untranslated region of miR-4731-5p. In summary, our research suggested that SENCR affects HA-VSMCs proliferation and migration via regulating the miR-4731-5p/FOXO3a pathway.     

脂多糖对离体培养大鼠血管平滑肌细胞增殖的影响

本研究观察到１０－７～１０－５ｋｇ／Ｌ脂多糖（ｌｉｐｏｐｏｌｙｓａｃｈａｒｉｄｅ,ＬＰＳ）可显著促进血管平滑肌细胞（ＶＳＭＣ）的增殖及ＤＮＡ的合成（Ｐ＜００５）。５×１０－４～１０－３ｋｇ／ＬＬＰＳ却抑制ＶＳＭＣ的增殖及ＤＮＡ的合成,降低其活力（Ｐ＜００１）,并呈时间依赖效应。一氧化氮合酶抑制剂ＮＮｉｔｒｏＬＡｒｇｉｎｉｎｅ（ＬＮＮＡ）可拮抗ＬＰＳ的抑制作用。大剂量ＬＰＳ作用组ＶＳＭＣ上清液中一氧化氮（ＮＯ）代谢产物ＮＯ－３和ＮＯ－２的含量与对照组相比显著增加（Ｐ＜００１）,４８ｈ组比２４ｈ组增加９１％,７２ｈ组比４８ｈ组增加４５％;同时,诱导性一氧化氮合酶（ｉｎｄｕｃｔｉｖｅｎｉｔｒｉｃｏｘｉｄｅｓｙｎｔｈａｓｅ,ｉＮＯＳ）免疫组化染色呈阳性。结果表明,低浓度ＬＰＳ促进ＶＳＭＣ增殖和ＤＮＡ合成,而高浓度ＬＰＳ却明显抑制ＶＳＭＣ增殖和ＤＮＡ合成,降低其活力。这种抑制作用可能与ＬＰＳ诱导ＶＳＭＣ产生的ＮＯ有关。     

Lin28a up‐regulation is associated with the formation of restenosis via promoting proliferation and migration of vascular smooth muscle cells

To explore the potential role of Lin28a in the development of restenosis after percutaneous transluminal angioplasty, double‐balloon injury surgery and mono‐balloon injury surgery were used to establish restenosis and atherosclerosis models, respectively, so as to better distinguish restenosis from atherosclerotic lesions. Immunohistochemical analysis revealed that significantly higher expression of Lin28a was observed in the iliac arteries of restenosis plaques than that of atherosclerosis plaques. Immunofluorescence studies showed the colocalization of Lin28a with α‐smooth muscle actin in restenosis plaques, rather than in atherosclerosis plaques, which suggested that Lin28a might be related to the unique behaviour of vascular smooth muscle cells (VSMCs) in restenosis. To further confirm above hypothesis, Lin28a expression was up‐regulated by transfection of Lenti‐Lin28a and inhibited by Lenti‐Lin28a‐shRNA transfection in cultured VSMCs, and then the proliferation and migration capability of VSMCs were detected by EdU and Transwell assays, respectively. Results showed that the proliferation and migration of VSMCs were significantly increased in accordance with the up‐regulation of Lin28a expression, while above behaviours of VSMCs were significantly suppressed after inhibiting the expression of Lin28a. In conclusion, the up‐regulation of Lin28a exerts its modulatory effect on VSMCs’ proliferation and migration, which may play a critical role in contributing to pathological formation of restenosis.     

Heartwood extract of Rhus verniciflua Stokes and its active constituent fisetin attenuate vasoconstriction through calcium-dependent mechanism in rat aorta

Rhus verniciflua Stokes (RVS) exert cardiovascular protective activity by promoting blood circulation, but its active ingredients and underlying mechanism have yet to be identified. This study investigated the vascular effects of RVS, focusing on vasoconstriction and smooth muscle Ca²⁺ signaling. RVS heartwood extract attenuated contraction of aortic rings induced by the vasoconstrictors serotonin and phenylephrine, and inhibited the Ca²⁺ signaling evoked by serotonin in vascular smooth muscle cells. Subsequent activity-guided fractionation identified fisetin as an active constituent exerting a Ca²⁺ inhibitory effect. Fisetin could inhibit major Ca²⁺ mobilization pathways including extracellular Ca²⁺ influx mediated by the L-type voltage-gated Ca²⁺ channel, Ca²⁺ release from the intracellular store and store-operated Ca²⁺ entry. In accordance with Ca²⁺ inhibitory effect, fisetin attenuated vasoconstriction by serotonin and phenylephrine. These results suggest that the anticontractile effect, which is presumably mediated by inhibition of Ca²⁺ signaling, may contribute to the improvement of blood circulation by RVS.