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Krüppel-like factor 4 mediates cellular migration and invasion by altering RhoA activity. kr<s:1> ppel样因子4通过改变RhoA活性介导细胞迁移和侵袭。
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2018-12-01 DOI: 10.1080/15419061.2018.1444034
Philip R Brauer, Jee Hun Kim, Humberto J Ochoa, Elizabeth R Stratton, Kathryn M Black, William Rosencrans, Eliza Stacey, Engda G Hagos

Kru¨ppel like factor 4 (KLF4) is a transcription factor that regulates genes related to differentiation and proliferation. KLF4 also plays a role in metastasis via epithelial to mesenchymal transition. Here, we investigate the function of Klf4 in migration and invasion using mouse embryonic fibroblasts and the RKO human colon cancer cell line. Compared to wild-type, cells lacking Klf4 exhibited increased migration-associated phenotypes. In addition, overexpression of Klf4 in Klf4-/- MEFs attenuated the presence of stress fibers to wild-type levels. An invasion assay suggested that lack of Klf4 resulted in increased invasive capacity. Finally, analysis of RhoA showed elevated RhoA activity in both RKO and MEF cells. Taken together, our results strongly support the novel role of KLF4 in a post-translational regulatory mechanism where KLF4 indirectly modulates the actin cytoskeleton morphology via activity of RhoA in order to inhibit cellular migration and invasion.

克隆样因子4 (KLF4)是一种调控细胞分化和增殖相关基因的转录因子。KLF4也在上皮细胞向间质细胞的转移中起作用。本研究利用小鼠胚胎成纤维细胞和RKO人结肠癌细胞系研究Klf4在迁移和侵袭中的功能。与野生型相比,缺乏Klf4的细胞表现出更多的迁移相关表型。此外,Klf4在Klf4-/- mef中的过表达将应力纤维的存在减弱到野生型水平。侵袭试验表明,缺乏Klf4导致侵袭能力增加。最后,RhoA分析显示RKO和MEF细胞中RhoA活性均升高。综上所述,我们的研究结果有力地支持了KLF4在翻译后调节机制中的新作用,其中KLF4通过RhoA的活性间接调节肌动蛋白细胞骨架形态,以抑制细胞迁移和侵袭。
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引用次数: 15
Establishment and characterization of a carcinoma-associated fibroblast cell line derived from a human salivary gland adenoid cystic carcinoma. 来源于人唾液腺腺样囊性癌的癌相关成纤维细胞系的建立和表征。
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2018-12-01 DOI: 10.1080/15419061.2018.1464000
Jing Kong, Han Zhao, Qianhui Shang, Zhifei Ma, Ni Kang, Junling Tan, Hyat Ahmed Ibrahim Alraimi, Tingjiao Liu

Salivary gland adenoid cystic carcinoma (SACC) is one of the most common malignancies in the oral and maxillofacial region. Carcinoma-associated fibroblast (CAF) is an important component in the tumor microenvironment and participates in SACC progression. In this study, we established a CAF cell line derived from a human SACC and named it CAF-SA. It was identified that CAF-SA expressed typical CAF biomarkers. Then, we studied the cellular communications between CAF-SA, tumor cells and endothelial cells. It was found that CAF-SA promoted the migration, invasion, and proliferation of SACC tumor cells in vitro. In addition, tube formation by endothelial cells was enhanced by CAF-SA. In vivo experiment showed that SACC cells formed larger xenografts in nude mice when they were transplanted with CAF-SA. Overall, we demonstrated that CAF-SA exhibited the most important defining feature of CAF by promoting cancer progression.

涎腺腺样囊性癌是口腔颌面部最常见的恶性肿瘤之一。癌相关成纤维细胞(CAF)是肿瘤微环境的重要组成部分,参与SACC的进展。在本研究中,我们从人SACC中提取了一株CAF细胞系,并将其命名为CAF- sa。鉴定出CAF- sa表达典型的CAF生物标志物。然后,我们研究了ca - sa与肿瘤细胞和内皮细胞之间的细胞通讯。体外实验发现,cafa - sa能促进SACC肿瘤细胞的迁移、侵袭和增殖。此外,ca - sa可促进内皮细胞形成小管。体内实验表明,用ca - sa移植的SACC细胞在裸鼠体内形成较大的异种移植物。总的来说,我们证明了CAF- sa通过促进癌症进展表现出CAF最重要的决定性特征。
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引用次数: 9
Fabrication of nanofiber coated with l-arginine via electrospinning technique: a novel nanomatrix to counter oxidative stress under crosstalk of co-cultured fibroblasts and satellite cells. 静电纺丝技术制备l-精氨酸包覆纳米纤维:一种抗成纤维细胞和卫星细胞串扰氧化应激的新型纳米基质。
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2018-12-01 Epub Date: 2018-09-05 DOI: 10.1080/15419061.2018.1493107
Sivakumar Allur Subramaniyan, Sunirmal Sheet, Saravanakumar Balasubramaniam, Swami Vetha Berwin Singh, Dileep Reddy Rampa, Sureshkumar Shanmugam, Da Rae Kang, Ho Sung Choe, Kwan Seob Shim

The objective of this study was to synthesize and characterize novel polyurethane (PU)-nanofiber coated with l-arginine by electrospinning technique. This study determined whether l-arginine conjugated with PU-nanofiber could stimulate cell proliferation and prevent H2O2-induced cell death in satellite cells co-cultured with fibroblasts isolated from Hanwoo (Korean native cattle). Our results showed that l-arginine conjugated with PU nanofiber could reduce cytotoxicity of co-cultured satellite cells. Protein expression levels of bcl-2 were significantly upregulated whereas those of caspase-3 and caspase-7 were significantly downregulated in co-culture of satellite cells compared to those of monoculture cells after treatment with PU-nanofiber coated with l-arginine and which confirmed by Confocal microscope. These results suggest that co-culture of satellite cells with fibroblasts might be able to counter oxidative stress through translocation/penetration of antioxidant, collagen, and molecules secreted to satellite cells. Therefore, this nanofiber might be useful as a wound dressing in animals to counter oxidative stresses.

采用静电纺丝技术合成了新型l-精氨酸包覆聚氨酯纳米纤维,并对其进行了表征。本研究确定了l-精氨酸结合pu纳米纤维是否能刺激卫星细胞增殖,防止h2o2诱导的卫星细胞死亡。卫星细胞与韩国本土牛分离的成纤维细胞共培养。结果表明,l-精氨酸与PU纳米纤维偶联可降低共培养卫星细胞的细胞毒性。共聚焦显微镜证实,经l-精氨酸包被pu纳米纤维处理后,卫星细胞共培养后bcl-2蛋白表达水平显著上调,caspase-3和caspase-7蛋白表达水平显著下调。这些结果表明,卫星细胞与成纤维细胞共培养可能能够通过向卫星细胞分泌的抗氧化剂、胶原蛋白和分子的易位/渗透来对抗氧化应激。因此,这种纳米纤维可以作为动物的伤口敷料来对抗氧化应激。
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引用次数: 8
Phosphatidylethanolamine Deficiency Impairs Escherichia coli Adhesion by Downregulating Lipopolysaccharide Synthesis, Which is Reversible by High Galactose/Lactose Cultivation 磷脂酰乙醇胺缺乏通过下调脂多糖合成影响大肠杆菌粘附,高半乳糖/乳糖培养可逆转这一过程
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2017-01-01 DOI: 10.1080/15419061.2017.1282468
Chuan Yu, Ming Li, Yanan Sun, Xingguo Wang, Yong Chen
Abstract As the initiation step of bacterial infection or biofouling, bacterial adhesion on cells or substrates is generally an optimal target for antibacterial design. Phosphatidylethanolamine (PE) is the principal phospholipid in bacteria, and its function in bacterial adhesion remains unclear. In this study, four E. coli strains including two PE-deficient mutants (PE−PC− and PE−PC+ strains) and two PE-containing wild-type controls (PE + PC− strains) were recruited to investigate the influence of PE deficiency on bacterial adhesion. We found that PE deficiency could impair E. coli adhesion on macrophages (human THP-1-derived and mouse RAW264.7 macrophages) or glass coverslips by downregulating lipopolysaccharide (LPS) biosynthesis, which could be reversible by high galactose/lactose but not glucose cultivation. The data imply that PE play important role in bacterial adhesion probably via affecting LPS biosynthesis and suggest that targeting PE biosynthesis is also a potential antibacterial strategy.
作为细菌感染或生物污垢的起始步骤,细菌在细胞或底物上的粘附通常是抗菌设计的最佳目标。磷脂酰乙醇胺(PE)是细菌体内的主要磷脂,其在细菌黏附中的作用尚不清楚。本研究利用4株大肠杆菌,包括2株PE- PC -和2株PE- PC+突变株和2株含PE的野生型对照(PE + PC -株),研究PE缺乏对细菌黏附的影响。我们发现,PE缺乏可以通过下调脂多糖(LPS)的生物合成来损害大肠杆菌在巨噬细胞(人thp -1来源和小鼠RAW264.7巨噬细胞)或玻璃盖上的粘附,这可以通过高半乳糖/乳糖培养逆转,但葡萄糖培养不可逆转。这些数据表明,PE可能通过影响LPS的生物合成在细菌粘附中发挥重要作用,并提示靶向PE的生物合成也是一种潜在的抗菌策略。
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引用次数: 5
Pannexin1 Single Nucleotide Polymorphism and Platelet Reactivity in a Cohort of Cardiovascular Patients Pannexin1单核苷酸多态性与心血管患者血小板反应性
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2017-01-01 DOI: 10.1080/15419061.2017.1282469
F. Stierlin, F. Molica, J. Reny, B. Kwak, P. Fontana
Abstract Pannexin1 (Panx1), a membrane channel-forming protein permitting the passage of small-sized molecules, such as ATP, is expressed in human platelets. Recently, we showed that inhibiting Panx1 affects collagen-induced platelet aggregation but not aggregation triggered by other agonists. We also found that a single nucleotide polymorphism (SNP; rs1138800) in the Panx1 gene encoded for a gain-of-function channel (Panx1-400C) and was associated with enhanced collagen-induced platelet reactivity. Here, we assessed the association of this SNP with platelet reactivity in a cohort of 758 stable cardiovascular patients from the ADRIE study treated with aspirin and/or clopidogrel. We found that presence of the Panx1-400C allele was not associated with platelet reactivity in stable cardiovascular patients, irrespective of the platelet aggregation agonist used (collagen, ADP or arachidonic acid) or the anti-platelet drug regimen. Moreover, the Panx1-400A > C SNP did also not affect the re-occurrence of cardiac ischemic events in the same stable cardiovascular patient cohort.
Pannexin1 (Panx1)是一种膜通道形成蛋白,允许小分子(如ATP)通过,在人血小板中表达。最近,我们发现抑制Panx1会影响胶原诱导的血小板聚集,但不会影响其他激动剂引发的血小板聚集。我们还发现单核苷酸多态性(SNP;rs1138800)在Panx1基因中编码一个功能获得通道(Panx1- 400c),并与胶原诱导的血小板反应性增强有关。在这项研究中,我们评估了来自ADRIE研究的758名接受阿司匹林和/或氯吡格雷治疗的稳定心血管患者的SNP与血小板反应性的关系。我们发现,在稳定性心血管患者中,Panx1-400C等位基因的存在与血小板反应性无关,与使用血小板聚集激动剂(胶原蛋白、ADP或花生四烯酸)或抗血小板药物方案无关。此外,Panx1-400A > C SNP也不影响相同稳定心血管患者队列中心脏缺血事件的再发生。
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引用次数: 11
Macrophages Aggravate Hypoxia-Induced Cardiac Microvascular Endothelial Cell Injury via Peroxynitrite: Protection by Tongxinluo 巨噬细胞通过过氧亚硝酸盐加重缺氧诱导的心肌微血管内皮细胞损伤:通心络的保护作用
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2015-11-02 DOI: 10.3109/15419061.2016.1155565
Xiujuan Wang, Kun Liu, Bin Li, Yanning Li, Kaiwei Ye, J. Qi, Yu Wang
Abstract Activated macrophages contribute to endothelial dysfunction; however, it is unclear how peroxynitrite contributes to macrophage-mediated human cardiac microvascular endothelial cell (HCMEC) injury in hypoxia. In macrophage-HCMEC co-cultures subjected to hypoxia, there was an increase in hypoxia-inducible factor (HIF)-1α, HIF-2α, inducible nitric oxide synthase (iNOS), endothelin-converting enzyme (ECE)-1 and cyclooxygenase-2 (COX-2), and concomitant decrease in prostacyclin synthase (PGIS). This was mimicked by a peroxynitrite donor and attenuated by its decomposition catalyst. Tongxinluo (TXL) could decrease HIF-2α, iNOS, ECE-1 and COX-2 and increase PGIS in a dose-dependent manner, with increase of vascular endothelial growth factor. The protein alterations verified the remarkably affected mRNAs, indicating that the effects of TXL were similar to but better than that of peroxynitrite decomposition catalyst. Furthermore, TXL inhibited macrophage-mediated nitrotyrosine accumulation and attenuated HCMEC injury. The results suggest that peroxynitrite contributes to macrophage-mediated HCMEC injury in hypoxia, and TXL attenuates HCMEC injury mainly by inhibiting peroxynitrite.
活化的巨噬细胞参与内皮功能障碍;然而,目前尚不清楚过氧亚硝酸盐在缺氧条件下如何导致巨噬细胞介导的人心脏微血管内皮细胞(HCMEC)损伤。在缺氧条件下,巨噬细胞- hcmec共培养中,缺氧诱导因子(HIF)-1α、HIF-2α、诱导型一氧化氮合酶(iNOS)、内皮素转换酶(ECE)-1和环氧化酶-2 (COX-2)升高,前列腺环素合成酶(PGIS)降低。这是由过氧亚硝酸盐供体模拟,并减弱其分解催化剂。通心络(TXL)能降低HIF-2α、iNOS、ce -1、COX-2,增加PGIS,并伴有血管内皮生长因子的升高,呈剂量依赖性。蛋白质的改变证实了显著影响的mrna,表明TXL的作用与过氧亚硝酸盐分解催化剂相似但优于过氧亚硝酸盐分解催化剂。此外,TXL抑制巨噬细胞介导的硝基酪氨酸积累,减轻HCMEC损伤。结果表明,过氧亚硝酸盐参与了缺氧条件下巨噬细胞介导的HCMEC损伤,TXL主要通过抑制过氧亚硝酸盐来减轻HCMEC损伤。
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引用次数: 10
Structure, Regulation and Function of Gap Junctions in Liver 肝脏间隙连接的结构、调节和功能
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2015-11-02 DOI: 10.3109/15419061.2016.1151875
Joost Willebrords, Sara Crespo Yanguas, M. Maes, E. Decrock, Nan Wang, L. Leybaert, T. C. da Silva, Isabel Veloso Alves Pereira, H. Jaeschke, B. Cogliati, M. Vinken
Abstract Gap junctions are a specialized group of cell-to-cell junctions that mediate direct intercellular communication between cells. They arise from the interaction of two hemichannels of adjacent cells, which in turn are composed of six connexin proteins. In liver, gap junctions are predominantly found in hepatocytes and play critical roles in virtually all phases of the hepatic life cycle, including cell growth, differentiation, liver-specific functionality and cell death. Liver gap junctions are directed through a broad variety of mechanisms ranging from epigenetic control of connexin expression to post-translational regulation of gap junction activity. This paper reviews established and novel aspects regarding the architecture, control and functional relevance of liver gap junctions.
间隙连接是一组特殊的细胞间连接,介导细胞间的直接通讯。它们产生于相邻细胞的两个半通道的相互作用,而这两个半通道又由六种连接蛋白组成。在肝脏中,间隙连接主要存在于肝细胞中,并在肝脏生命周期的几乎所有阶段发挥关键作用,包括细胞生长、分化、肝脏特异性功能和细胞死亡。肝间隙连接通过多种机制进行调控,从连接蛋白表达的表观遗传控制到间隙连接活性的翻译后调控。本文综述了关于肝间隙连接的结构、控制和功能相关性的现有和新的方面。
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引用次数: 19
Regulation of Endothelial Cell Adherence and Elastic Modulus by Substrate Stiffness 基质刚度对内皮细胞粘附和弹性模量的调节
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2015-11-02 DOI: 10.1080/15419061.2016.1265949
Sharareh Jalali, M. Tafazzoli-Shadpour, N. Haghighipour, Ramin Omidvar, F. Safshekan
Abstract Although substrate stiffness has been previously reported to affect various cellular aspects, such as morphology, migration, viability, growth, and cytoskeletal structure, its influence on cell adherence has not been well examined. Here, we prepared three soft, medium, and hard polyacrylamide (PAAM) substrates and utilized AFM to study substrate elasticity and also the adhesion and mechanical properties of endothelial cells in response to changing substrate stiffness. Maximum detachment force and cell stiffness were increased with increasing substrate stiffness. Maximum detachment force values were 0.28 ± 0.14, 0.94 ± 0.27, and 1.99 ± 0.59 nN while Young’s moduli of cells were 218.85 ± 38.73, 385.58 ± 131.67, and 933.20 ± 428.92 Pa for soft, medium, and hard substrates, respectively. Human umbilical vein endothelial cells (HUVECs) showed round to more spread shapes on soft to hard substrates, with the most organized and elongated actin structure on the hard hydrogel. Our results confirm the importance of substrate stiffness in regulating cell mechanics and adhesion for a successful cell therapy.
虽然之前有报道称底物刚度会影响细胞的各个方面,如形态、迁移、活力、生长和细胞骨架结构,但其对细胞粘附的影响尚未得到很好的研究。在这里,我们制备了三种软、中、硬聚丙烯酰胺(PAAM)基质,并利用AFM研究了基质弹性以及内皮细胞对基质刚度变化的粘附和力学性能。最大剥离力和细胞刚度随基质刚度的增加而增加。软、中、硬基质的最大分离力分别为0.28±0.14、0.94±0.27和1.99±0.59 nN,细胞的杨氏模量分别为218.85±38.73、385.58±131.67和933.20±428.92 Pa。人脐静脉内皮细胞(HUVECs)在软硬基质上呈圆形或更分散的形状,其中硬水凝胶上的肌动蛋白结构最整齐、细长。我们的研究结果证实了基质硬度在调节细胞力学和粘附方面的重要性,这对于成功的细胞治疗至关重要。
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引用次数: 46
Cytokine Release Patterns in Mixed Lymphocyte Culture (MLC) of T-Cells with Dendritic Cells (DC) Generated from AML Blasts Contribute to Predict anti-Leukaemic T-Cell Reactions and Patients' Response to Immunotherapy. 混合淋巴细胞培养(MLC) t细胞与AML母细胞产生的树突状细胞(DC)的细胞因子释放模式有助于预测抗白血病t细胞反应和患者对免疫治疗的反应。
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2015-04-01 Epub Date: 2016-09-07 DOI: 10.1080/15419061.2016.1223634
Dorothea Fischbacher, Marion Merle, Anja Liepert, Christine Grabrucker, Tanja Kroell, Andreas Kremser, Julia Dreyßig, Markus Freudenreich, Friedhelm Schuster, Arndt Borkhardt, Doris Kraemer, Claus-Henning Koehne, Hans-Jochem Kolb, Christoph Schmid, Helga Maria Schmetzer

To enlighten interactions between autologous, allogeneic or T-cells from patients after stem cell transplantation with leukaemia-derived-dendritic-cells containing dendritic cells or blast containing mononuclear cells (n = 21, respectively), we determined cytokine-concentrations (interleukin 2, 4, 6, 10, tumor-necrosis-factor-α, interferon-γ) in supernatants of mixed-lymphocyte-culture and in serum (n = 16) of 20 patients with acute myeloid leukaemia and three patients with myelodysplastic syndromes by cytometric-bead-assay. We correlated our data with lytic capabilities of stimulated T-cells in a fluorolysis-assay and clinical data: Dendritic-cell-/mononuclear-cell-stimulation of T-cells resulted in increased cytokine-levels in culture-medium compared to serum. There were no significant differences between cytokine-patterns of cases with/without lytic T-cell-activity, response to immunotherapy (stem cell transplantation/donor-lymphocyte-infusion) or graft-versus-host-disease. However, some predictive cytokine-cut-off-values for antileukaemic T-cell-activity, patients' response to immunotherapy and graft-versus-host-disease could be defined. Cytokine-profiles alone, without functional assays, are no useful tool to predict antileukaemic T-cell-function, although they can indicate lytic T-cell-activity, patients' response to immunotherapy and graft-versus-host-disease.

为了揭示干细胞移植后患者自体、异体或t细胞与含有树突状细胞的白血病衍生树突状细胞或含有单个核细胞的母细胞(n = 21)之间的相互作用,我们测定了细胞因子浓度(白细胞介素2、4、6、10、肿瘤坏死因子-α、20例急性髓性白血病患者和3例骨髓增生异常综合征患者的混合淋巴细胞培养上清和血清(n = 16)中干扰素-γ的含量。我们将我们的数据与受刺激的t细胞在荧光溶解试验中的溶解能力和临床数据相关联:与血清相比,树突状细胞/单核细胞对t细胞的刺激导致培养基中细胞因子水平升高。在有/没有溶解性t细胞活性、对免疫治疗(干细胞移植/供体淋巴细胞输注)或移植物抗宿主病的病例中,细胞因子模式没有显著差异。然而,抗白血病t细胞活性、患者对免疫治疗的反应和移植物抗宿主病的一些预测性细胞因子切断值可以被定义。单独的细胞因子谱,没有功能测定,并不是预测抗白血病t细胞功能的有用工具,尽管它们可以指示溶解性t细胞活性、患者对免疫治疗的反应和移植物抗宿主病。
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引用次数: 7
Glycolaldehyde-derived advanced glycation end products (glycol-AGEs)-induced vascular smooth muscle cell dysfunction is regulated by the AGES-receptor (RAGE) axis in endothelium. 乙醇醛衍生的晚期糖基化终产物(glycolage)诱导的血管平滑肌细胞功能障碍受内皮中ages受体(RAGE)轴的调控。
Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2015-04-01 Epub Date: 2016-09-07 DOI: 10.1080/15419061.2016.1225196
Mi-Hyun Nam, Won-Rak Son, Young Sik Lee, Kwang-Won Lee

Advanced glycation end-products (AGEs) are involved in the development of vascular smooth muscle cell (VSMC) dysfunction and the progression of atherosclerosis. However, AGEs may indirectly affect VSMCs via AGEs-induced signal transduction between monocytes and human umbilical endothelial cells (HUVECs), rather than having a direct influence. This study was designed to elucidate the signaling pathway underlying AGEs-RAGE axis influence on VSMC dysfunction using a co-culture system with monocytes, HUVECs and VSMCs. AGEs stimulated production of reactive oxygen species and pro-inflammatory mediators such as tumor necrosis factor-α and interleukin-1β via extracellular-signal-regulated kinases phosphorylation and nuclear factor-κB activation in HUVECs. It was observed that AGEs-induced pro-inflammatory cytokines increase VSMC proliferation, inflammation and vascular remodeling in the co-culture system. This result implies that RAGE plays a role in AGEs-induced VSMC dysfunction. We suggest that the regulation of signal transduction via the AGEs-RAGE axis in the endothelium can be a therapeutic target for preventing atherosclerosis.

晚期糖基化终产物(AGEs)参与血管平滑肌细胞(VSMC)功能障碍的发生和动脉粥样硬化的进展。然而,AGEs可能通过诱导单核细胞和人脐内皮细胞(HUVECs)之间的信号转导间接影响VSMCs,而不是直接影响。本研究旨在通过单核细胞、HUVECs和VSMC共培养系统,阐明AGEs-RAGE轴影响VSMC功能障碍的信号通路。AGEs通过细胞外信号调节的激酶磷酸化和核因子-κB激活,刺激HUVECs中活性氧和促炎介质如肿瘤坏死因子-α和白细胞介素-1β的产生。研究发现,ages诱导的促炎细胞因子增加了共培养系统中VSMC的增殖、炎症和血管重构。这一结果提示RAGE在ages诱导的VSMC功能障碍中起作用。我们认为通过内皮细胞的age - rage轴调节信号转导可能是预防动脉粥样硬化的治疗靶点。
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引用次数: 30
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Cell Communication and Adhesion
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