• 【角质细胞迁移和肽生长因子: PDGF,bFGF,EGF,igf-i,aFGF和TGF-β 对胶原蛋白凝胶中人角质细胞迁移的影响。】 复制标题 收藏 收藏
    DOI:10.1076/ceyr.16.6.605.5081 复制DOI
    作者列表:Andresen JL,Ledet T,Ehlers N
    BACKGROUND & AIMS: PURPOSE:Peptide growth factors are known accelerators of corneal wound healing, probably mediated through increased proliferation of the cells; however, information about their effect on keratocyte motility is lacking. The influence of peptide growth factors on keratocyte migratory activity was investigated, using the following growth factors: platelet derived growth factor (PDGF-BB), epidermal growth factor (EGF), acidic fibroblast growth factor (aFGF), basic fibroblast growth factor (bFGF), insulin-like growth factor-I (IGF-I) and transforming growth factor-beta-1 (TGF-beta 1).

    METHODS:Keratocytes were seeded on gels of type 1 collagen, growth factor added, and the cells left to migrate for 72 hours. Subsequently, the number of keratocytes at the different levels in the collagen gel was evaluated by optically sectioning the gel at 20 microns, intervals, with an inverted phase contrast microscope.

    RESULTS:PDGF, EGF and bFGF at 10 ng/ml, all increased the number of keratocytes at the different levels of the gel as compared to a non-stimulated control (p < 0.05 or p < 0.01, students t-test). TGF-beta proved to be a strong inhibitor of keratocyte migration, decreasing the number of keratocytes observed at every level in the gel (p < 0.05 and p < 0.01, students t-test), whereas no effect of IGF-I and aFGF was found. During the 72 hours of migration, no contraction of the collagen gels was observed. Autoradiography of histological sections of the gels showed that during the 72-hour period only TGF-beta and 10% fetal bovine serum induced an increase in keratocyte proliferation.

    CONCLUSION:PDGF, EGF and bFGF increase keratocyte migration, independent of proliferation in a collagen gel invasion assay and might promote corneal wound healing, not only by increasing cell proliferation, but also through increased motility.

    背景与目标: 目的 : 肽生长因子是已知的角膜伤口愈合的促进剂,可能是通过细胞增殖增加介导的; 但是,缺乏有关它们对角质细胞运动的影响的信息。用以下生长因子: 血小板衍生生长因子 (pdgf-bb),表皮生长因子 (EGF),酸性成纤维细胞生长因子 (aFGF),碱性成纤维细胞生长因子 (bFGF),胰岛素样生长因子-I (igf-i) 和转化生长因子-β1 (tgf-β1)。
    方法 : 将角质细胞接种在1型胶原蛋白的凝胶上,添加生长因子,细胞迁移72小时。随后,通过用倒置相差显微镜以20微米的间隔光学切片来评估胶原蛋白凝胶中不同水平的角质形成细胞的数量。
    结果 :PDGF,EGF和bFGF为10 ng/ml,与未刺激的对照相比,在凝胶的不同水平下,所有这些都增加了角质细胞的数量 (p <0.05或p <0.01,学生t检验)。TGF-β 被证明是角质细胞迁移的强抑制剂,减少了在凝胶中每个水平观察到的角质细胞的数量 (p <0.05和p <0.01,学生t检验),而没有发现igf-i和aFGF的作用。在迁移的72小时内,未观察到胶原蛋白凝胶的收缩。凝胶组织学切片的放射自显影显示,在72小时内,只有TGF-β 和10% 胎牛血清诱导角质细胞增殖增加。
    结论 :PDGF,EGF和bFGF增加角质细胞迁移,在胶原蛋白凝胶侵袭试验中独立于增殖,并且可能不仅通过增加细胞增殖,而且通过增加运动性来促进角膜伤口愈合。
  • 【抗原特异性T细胞克隆在胶原疾病中的意义: 用新型T细胞克隆性评估系统分析。】 复制标题 收藏 收藏
    DOI:10.2169/internalmedicine.36.242 复制DOI
    作者列表:Yamamoto K
    BACKGROUND & AIMS: The involvement of antigen-specific T cells in the pathogenesis of collagen diseases is still controversial. The final stages of collagen diseases are usually characterized by the dominance of inflammation. Therefore, antigen non-specific factors, such as inflammatory cytokines, probably play an important role in this process. On the other hand, the methods available to analyze the antigen-specific aspects of the immune response are still limited. Here we review our novel system of T cell clonality analysis based on the idea that activated antigen-specific T cells should form accumulating clones among the lymphocyte population. Using this method, dynamic changes of clonal accumulation of T cells could be evaluated during antigenic stimulation in vivo and in vitro. The significance of antigen-specific T cell clones in collagen diseases is discussed using data obtained from patients with rheumatoid arthritis and systemic lupus erythematosus.

    背景与目标: 抗原特异性T细胞参与胶原疾病的发病机理仍存在争议。胶原蛋白疾病的最后阶段通常以炎症为主。因此,抗原非特异性因子 (如炎性细胞因子) 可能在这一过程中起重要作用。另一方面,用于分析免疫应答的抗原特异性方面的方法仍然有限。在这里,我们基于激活的抗原特异性T细胞应在淋巴细胞群体中形成累积克隆的想法,回顾了我们的新型T细胞克隆分析系统。使用该方法,可以在体内和体外评估抗原刺激过程中T细胞克隆积累的动态变化。使用类风湿关节炎和系统性红斑狼疮患者获得的数据讨论了抗原特异性T细胞克隆在胶原疾病中的意义。
  • 【槲寄生制剂 (Iscador) 在三维胶原蛋白基质系统中诱导T淋巴细胞运动的供体依赖性和剂量依赖性变化。】 复制标题 收藏 收藏
    DOI:10.1097/00001813-199704001-00014 复制DOI
    作者列表:Nikolai G,Friedl P,Werner M,Zänker KS
    BACKGROUND & AIMS: :Controlled activation of non-specific and specific immune defence mechanisms can beneficially manipulate the host's ability to attack malignant cells. In this context, migration and tissue distribution of immunocompetent cells may be prerequisites for an efficient immune surveillance. The effect of various non-cytotoxic concentrations of the Viscum album L. (mistletoe) preparation Iscadore QuFrF on the locomotory activity of immunomagnetically isolated human CD4+ and CD8+ T lymphocytes from healthy donors was investigated. Cellular migration was examined within a three-dimensional collagen matrix. Donor-dependent variations in baseline activities of spontaneously locomoting T cells were accompanied by individual response patterns of T cells from different donors in the presence of various concentrations of mistletoe preparation (0.25-2.5 micrograms/ml). Using the three-dimensional collagen matrix assay an induction of locomotory activity was detected in a highly reproducible fashion although the optimal concentration of mistletoe preparation and the time point of maximal response were individual for each donor. Our data suggest that the direct stimulation of T-cell migration by mistletoe components may modulate the system of immune surveillance and recognition in patients under mistletoe therapy.
    背景与目标: : 非特异性和特异性免疫防御机制的受控激活可以有利地操纵宿主攻击恶性细胞的能力。在这种情况下,免疫活性细胞的迁移和组织分布可能是有效免疫监视的先决条件。Viscum专辑L的各种非细胞毒性浓度的影响。(槲寄生) 制剂Iscadore QuFrF对来自健康供体的免疫磁分离的人CD4和CD8 T淋巴细胞的运动活性进行了研究。在三维胶原蛋白基质中检查细胞迁移。在存在各种浓度的槲寄生制剂 (0.25-2.5微克/毫升) 的情况下,自发运动的T细胞的基线活性的供体依赖性变化伴随着来自不同供体的T细胞的个体反应模式。使用三维胶原基质测定法,尽管槲寄生制剂的最佳浓度和最大反应的时间点对于每个供体都是单独的,但仍以高度可重复性的方式检测到运动活性的诱导。我们的数据表明,槲寄生成分对T细胞迁移的直接刺激可能会调节槲寄生治疗患者的免疫监视和识别系统。
  • 【纤维连接蛋白促进人角膜成纤维细胞介导的胶原凝胶收缩。】 复制标题 收藏 收藏
    DOI:10.1016/j.exer.2006.06.008 复制DOI
    作者列表:Liu Y,Yanai R,Lu Y,Kimura K,Nishida T
    BACKGROUND & AIMS: :Collagen contraction mediated by corneal fibroblasts (CFs) is implicated in the maintenance of corneal shape. Given that fibronectin is expressed at sites of corneal stromal wounding, we investigated the effect of fibronectin on CF-mediated collagen gel contraction. Human CFs were cultured in a three-dimensional gel of type I collagen in the absence or presence of various extracellular matrix (ECM) components. The contraction of collagen gels mediated by CFs was evaluated by measurement of changes in gel diameter. The formation of stress fibers and focal adhesions in CFs was examined by fluorescence microscopy. The abundance of paxillin, phosphorylated paxillin, integrins alpha5, beta1, and alpha2, and alpha-smooth muscle actin in CFs was examined by immunoblot analysis. Fibronectin promoted CF-mediated collagen gel contraction in a concentration- and time-dependent manner. Other ECM proteins or glycosaminoglycans did not exhibit such an effect. Fibronectin also induced cell spreading, the formation of stress fibers, and the establishment of focal adhesions containing paxillin in CFs cultured in three-dimensional collagen gels. In addition, it increased the amounts of paxillin, phosphorylated paxillin, and integrins alpha5 and beta1 in these cells. The expression of integrin alpha2 and alpha-smooth muscle actin was not affected by fibronectin, however. Furthermore, the peptide GRGDSP (an antagonist of fibronectin receptors) blocked the stimulatory effect of fibronectin on CF-mediated collagen gel contraction. These results suggest that fibronectin promoted CF-mediated collagen gel contraction in a manner dependent on the formation of stress fibers and focal adhesions, the activation of paxillin, and the up-regulation of integrin alpha5beta1. Fibronectin may therefore contribute to the maintenance of corneal shape by CFs during the healing of stromal wounds.
    背景与目标: 角膜成纤维细胞 (CFs) 介导的胶原收缩与角膜形状的维持有关。鉴于纤连蛋白在角膜基质损伤部位表达,我们研究了纤连蛋白对CF介导的胶原凝胶收缩的影响。在不存在或存在各种细胞外基质 (ECM) 成分的情况下,在I型胶原蛋白的三维凝胶中培养人CFs。通过测量凝胶直径的变化来评估CFs介导的胶原蛋白凝胶的收缩。通过荧光显微镜检查CFs中应力纤维和粘着斑的形成。通过免疫印迹分析检查CFs中桩蛋白,磷酸化桩蛋白,整联蛋白 α5,β1和 α2以及 α-平滑肌肌动蛋白的丰度。纤连蛋白以浓度和时间依赖性方式促进CF介导的胶原蛋白凝胶收缩。其他ECM蛋白或糖胺聚糖没有表现出这种作用。纤连蛋白还诱导细胞扩散,应力纤维的形成以及在三维胶原蛋白凝胶中培养的CFs中建立含有桩蛋白的粘着斑。此外,它增加了这些细胞中的桩蛋白,磷酸化的桩蛋白以及整合素 α5和 β1的量。然而,整合素 α2和 α-平滑肌肌动蛋白的表达不受纤连蛋白的影响。此外,肽GRGDSP (纤连蛋白受体的拮抗剂) 阻断了纤连蛋白对CF介导的胶原蛋白凝胶收缩的刺激作用。这些结果表明,纤连蛋白以取决于应力纤维和粘着斑的形成,桩蛋白的活化以及整联蛋白alpha5beta1的上调的方式促进CF介导的胶原蛋白凝胶收缩。因此,纤连蛋白可能有助于CFs在基质伤口愈合过程中维持角膜形状。
  • 【粘附性糖蛋白纤连蛋白的胶原结合片段的鉴定和隔离。】 复制标题 收藏 收藏
    DOI:10.1073/pnas.76.3.1160 复制DOI
    作者列表:Hahn LH,Yamada KM
    BACKGROUND & AIMS: :We have identified and purified a polypeptide region containing the collagen-binding site of the adhesive glycoprotein fibronectin. Chicken cellular fibronectin isolated from cultured embryonic fibroblasts was permitted to bind to gelatin coupled to agarose beads and was then digested extensively with chymotrypsin. A prominent 40,000-dalton fragment of fibronectin consisting of a single polypeptide chain was detected by sodium dodecyl sulfate/polyacrylamide gel electrophoresis of material remaining bound to the gelatin-agarose. This fragment appeared within 10 min after the digestion was initiated and persisted for more than 20 hr. This proteolytic fragment was isolated in electrophoretically pure form and retained its affinity for collagen. Plasma fibronectins from chicken and human blood also contained collagen-binding proteolytic fragments of similar size. This finding suggest that the collagen-binding sites of cellular and plasma fibronectins are homologous.
    背景与目标: : 我们已经鉴定并纯化了含有粘附糖蛋白纤连蛋白的胶原结合位点的多肽区域。允许从培养的胚胎成纤维细胞中分离出的鸡细胞纤连蛋白与琼脂糖珠偶联的明胶结合,然后用胰凝乳蛋白酶广泛消化。通过与明胶-琼脂糖结合的材料的十二烷基硫酸钠/聚丙烯酰胺凝胶电泳检测到由单个多肽链组成的纤连蛋白的突出的40,000-道尔顿片段。该片段在消化开始后10分钟内出现,并持续20小时以上。该蛋白水解片段以电泳纯的形式分离,并保留了其对胶原蛋白的亲和力。来自鸡和人血的血浆纤连蛋白也包含大小相似的胶原蛋白结合蛋白水解片段。这一发现表明细胞和血浆纤连蛋白的胶原结合位点是同源的。
  • 【Tyro3,Axl和Mer酪氨酸激酶激动剂对胶原诱导的关节炎的治疗效果。】 复制标题 收藏 收藏
    DOI:10.1002/art.37786 复制DOI
    作者列表:van den Brand BT,Abdollahi-Roodsaz S,Vermeij EA,Bennink MB,Arntz OJ,Rothlin CV,van den Berg WB,van de Loo FA
    BACKGROUND & AIMS: OBJECTIVE:Hyperactivation of innate immunity by Toll-like receptors (TLRs) can contribute to the development of autoinflammatory or autoimmune diseases. This study evaluated the activation of Tyro3, Axl, Mer (TAM) receptors, physiologic negative regulators of TLRs, by their agonists, growth arrest-specific protein 6 (GAS-6) and protein S, in the prevention of collagen-induced arthritis (CIA). METHODS:Adenoviruses overexpressing GAS-6 and protein S were injected intravenously or intraarticularly into mice during CIA. Splenic T helper cell subsets from intravenously injected mice were studied by flow cytometry, and the knee joints of mice injected intravenously and intraarticularly were assessed histologically. Synovium from mice injected intraarticularly was evaluated for cytokine and suppressor of cytokine signaling (SOCS) expression. RESULTS:Protein S significantly reduced ankle joint swelling when overexpressed systemically. Further analysis of knee joints revealed a moderate reduction in pathologic changes in the joint and a significant reduction in the number of splenic Th1 cells when protein S was overexpressed systemically. Local overexpression of GAS-6 decreased joint inflammation and joint pathology. Protein S treatment showed a similar trend of protection. Consistently, GAS-6 and protein S reduced cytokine production in the synovium. Moreover, levels of messenger RNA for interleukin-12 (IL-12) and IL-23 were reduced by GAS-6 and protein S treatment, with a corresponding decrease in the production of interferon-γ and IL-17. TAM ligand overexpression was associated with an increase in SOCS-3 levels, which likely contributed to the amelioration of arthritis. CONCLUSION:This study provides the first evidence that TAM receptor stimulation by GAS-6 and protein S can be used to ameliorate arthritis when applied systemically or locally. TAM receptor stimulation limits proinflammatory signaling and adaptive immunity. This pathway provides a novel strategy by which to combat rheumatoid arthritis.
    背景与目标:
  • 【胶原: 酰胺I带手性二阶光学非线性的结构起源。】 复制标题 收藏 收藏
    DOI:10.1016/j.bpj.2012.10.017 复制DOI
    作者列表:Reiser KM,McCourt AB,Yankelevich DR,Knoesen A
    BACKGROUND & AIMS: :The molecular basis of nonlinear optical (NLO) chiral effects in the amide I region of type I collagen was investigated using sum-frequency generation vibrational spectroscopy; chiral and achiral tensor elements were separated using different input/output beam polarization conditions. Spectra were obtained from native rat tail tendon (RTT) collagen and from cholesteric liquid crystal-like (LC) type I collagen films. Although RTT and LC collagen both possess long-range order, LC collagen lacks the complex hierarchical organization of RTT collagen. Their spectra were compared to assess the role of such organization in NLO chirality. No significant differences were observed between RTT and LC with respect to chiral or achiral spectra. These findings suggest that amide I NLO chiral effects in type I collagen assemblies arise predominantly from the chiral organization of amide chromophores within individual collagen molecules, rather than from supramolecular structures. The study suggests that sum-frequency generation vibrational spectroscopy may be uniquely valuable in exploring fundamental aspects of chiral nonlinearity in complex macromolecular structures.
    背景与目标: : 使用和频产生振动光谱研究了I型胶原的酰胺I区域中非线性光学 (NLO) 手性效应的分子基础; 使用不同的输入/输出光束偏振条件分离手性和非手性张量元素。光谱是从天然的大鼠尾腱 (RTT) 胶原蛋白和胆甾型液晶样 (LC) I型胶原蛋白膜获得的。尽管RTT和LC胶原蛋白都具有长程顺序,但LC胶原蛋白缺乏RTT胶原蛋白的复杂层次组织。比较了它们的光谱,以评估这种组织在NLO手性中的作用。在手性或无手性光谱方面,RTT和LC之间未观察到显着差异。这些发现表明,I型胶原蛋白组装体中的酰胺I NLO手性作用主要来自单个胶原蛋白分子中酰胺发色团的手性组织,而不是超分子结构。研究表明,和频产生振动光谱在探索复杂大分子结构中手性非线性的基本方面可能具有独特的价值。
  • 【使用与I型胶原蛋白的C-端肽的8个氨基酸序列的异构化形式反应的抗体测量血清中的骨降解产物。】 复制标题 收藏 收藏
    DOI:10.1359/jbmr.1997.12.7.1028 复制DOI
    作者列表:Bonde M,Garnero P,Fledelius C,Qvist P,Delmas PD,Christiansen C
    BACKGROUND & AIMS: An enzyme-linked immunosorbent assay for measuring type I collagen degradation products in serum (S-ELISA) was developed. The assay uses a high affinity polyclonal antibody which reacts with an isomerized form of an 8 amino acid sequence of the C-telopeptides of type I collagen (EKAHD-beta-GGR). Cross-reactivity to a nonisomerized synthetic peptide form of the 8 amino acid sequence is less than 0.2%. Values obtained in a group of premenopausal women (age, 33.3 +/- 3.11 years) were 69 +/- 24 ng/ml(n = 22). In a group of early postmenopausal women (age, 51.8 +/- 1.88 years) values obtained were 125 +/- 43 ng/ml (n = 46), which represents an increase of 81% (p < 0.001). Values found in untreated patients with Paget's disease were 234 +/- 95 ng/ml (n = 15), and for primary hyperparathyroidism we found 335 +/- 82 ng/ml (n = 10). Intravenous administration of a bisphosphonate (Pamidronate) to Paget's disease patients for 3 days was reflected in the S-ELISA by a decrease in the values of 55% when compared with values before treatment (n = 15). Following treatment with another bisphosphonate (Alendronate) for 6 months, values were decreased to 48 +/- 19 ng/ml (n = 12), which corresponds to a 62% decrease. Clinical results presented in this context support that the assay is a sensitive and specific index of bone resorption. It may, therefore, prove useful in the follow up of treatment of patients with metabolic bone diseases and in the clinical investigation of osteoporosis.

    背景与目标: 开发了一种用于测量血清中I型胶原蛋白降解产物的酶联免疫吸附测定法 (S-ELISA)。该测定使用高亲和力多克隆抗体,该抗体与I型胶原蛋白 (EKAHD-β-GGR) 的C-端肽的8个氨基酸序列的异构化形式反应。对8个氨基酸序列的非异构化合成肽形式的交叉反应性小于0.2%。在一组绝经前妇女 (年龄,33.3 +/- 3.11岁) 中获得的值为69 +/- 24 ng/ml(n = 22)。在一组绝经后早期妇女 (年龄,51.8 +/- 1.88岁) 中,获得的值为125 +/- 43 ng/ml (n = 46),这表示增加了81% (p <0.001)。在未经治疗的Paget病患者中发现的值为234/- 95 ng/ml (n = 15),对于原发性甲状旁腺功能亢进,我们发现335/- 82 ng/ml (n = 10)。与治疗前的值相比 (n = 15),在S-ELISA中反映了将双膦酸盐 (帕米膦酸盐) 静脉给药至Paget病患者3天的55% 值降低。用另一种双膦酸盐 (阿仑膦酸盐) 治疗6个月后,值降低至48 +/- 19 ng/ml (n = 12),这对应于62% 降低。在这种情况下提出的临床结果支持该测定是骨吸收的敏感且特异性的指标。因此,它可能对代谢性骨病患者的治疗随访和骨质疏松症的临床研究有用。
  • 【瘦素的生理浓度增加了非永生化人肝星状细胞产生的胶原蛋白。】 复制标题 收藏 收藏
    DOI:10.1016/j.metabol.2006.05.016 复制DOI
    作者列表:Choudhury J,Mirshahi F,Murthy KS,Yager DR,Sanyal AJ
    BACKGROUND & AIMS: :The effects of leptin, in concentrations seen in obesity, on collagen production and turnover in non-immortalized human hepatic stellate cell (HSC), were unknown. The profibrogenic effects of leptin in these cells were studied. Hepatic stellate cells were obtained from resected livers. Collagen I/III gene expression and protein production were measured by quantitative real-time polymerase chain reaction and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, respectively. The signal transduction pathways involved were evaluated by specific blockers of the phosphatidylinositol 3-kinase (PI3K), mitogen-activated protein kinase kinase (MEK), and Janus kinase 2 (JAK2). The effects on matrix metalloproteinase 1 (MMP-1) and tissue inhibitor of metalloproteinase 1 (TIMP-1) were assessed by their gene transcript levels, collagenolytic activity of cell culture supernatants, and MMP-1 protein levels. At concentrations seen in nonobese individuals ([leptin] < 10 ng/mL), leptin did not affect collagen production. At concentrations seen in obesity (30-50 ng/mL), leptin increased collagen I and III messenger RNA (mRNA) transcript levels by 286% +/- 55% (P < .001) and 167% +/- 62% (P < .007) and protein production by 45.8% +/- .02% and 84.39% +/- .01%, respectively. These effects were blocked by JAK2 inhibition as well as PI3K inhibition. Although MEK inhibition blocked leptin-induced procollagen I and III mRNA levels, there were no significant effects on collagen I and III protein levels. Leptin (10-50 ng/mL) had no significant effects on MMP-1 or TIMP-1 mRNA levels, collagenolytic activity, or MMP-1 protein levels. In conclusion, leptin, at levels seen in obese individuals, produces an increase in collagen production by HSC acting through the JAK and PI3K pathways. At these concentrations, leptin does not affect MMP-1 or TIMP-1 expression or collagenolytic activity of HSC.
    背景与目标: : 瘦素 (在肥胖中观察到的浓度) 对非永生化人肝星状细胞 (HSC) 中胶原蛋白产生和周转的影响尚不清楚。研究了瘦素在这些细胞中的促纤维化作用。从切除的肝脏中获得肝星状细胞。通过定量实时聚合酶链反应和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分别测量胶原蛋白I/III基因表达和蛋白质产生。通过磷脂酰肌醇3-激酶 (PI3K),丝裂原活化蛋白激酶激酶 (MEK) 和Janus激酶2 (JAK2) 的特异性阻滞剂评估了所涉及的信号转导途径。通过其基因转录水平,细胞培养上清液的胶原分解活性和MMP-1蛋白水平评估了对基质金属蛋白酶1 (MMP-1) 和组织金属蛋白酶1 (TIMP-1) 的影响。在非肥胖个体中观察到的浓度 ([leptin] < 10 ng/mL),leptin不会影响胶原蛋白的产生。在肥胖中观察到的浓度 (30-50 ng/mL),瘦素通过286% +/- 55% (P < .001) 和167% +/- 62% (P < .007) 增加胶原蛋白I和III信使RNA (mRNA) 转录水平,并通过45.8% +/- .02% 和84.39% +/- .01% 增加蛋白质生产。这些作用被JAK2抑制和PI3K抑制所阻断。尽管MEK抑制抑制了瘦素诱导的胶原蛋白I和III mRNA水平,但对胶原蛋白I和III蛋白水平没有显着影响。瘦素 (10-50ng/mL) 对MMP-1或TIMP-1 mRNA水平,胶原蛋白分解活性或MMP-1蛋白水平没有显着影响。总之,在肥胖个体中观察到的瘦素水平,通过HSC通过JAK和PI3K途径起作用而增加胶原蛋白的产生。在这些浓度下,瘦素不影响HSC的MMP-1或TIMP-1表达或胶原分解活性。
  • 【胶原刚度调节细胞收缩和基质重塑基因表达。】 复制标题 收藏 收藏
    DOI:10.1002/jbm.a.31423 复制DOI
    作者列表:Karamichos D,Brown RA,Mudera V
    BACKGROUND & AIMS: :Cell-level mechanical and 3D spatial cues are essential to the organization and architecture of new tissues that form during growth, repair or in bioreactors. Fibroblast-seeded 3D collagen constructs have been used as bioartifical extracellular matrix (ECM) providing a 3D environment to embedded resident cells. As cells attach to scaffold fibrils, they generate quantifiable contractile forces which depend on cell type, cell attachment, cell density, growth factors, and matrix stiffness. The aim of this study was to quantify the cytomechanical and molecular responses of human dermal (HDF) and neonatal foreskin fibroblasts (HNFF) seeded in constructs of increased stiffness. We also tested the effect of blocking early attachment using serum starvation on these outputs. Constructs were placed under uniaxial strains of 0-10% to increase scaffold stiffness, prior to gel contraction, and force generation was monitored using a tensional culture force monitor (t-CFM). Increased matrix stiffness reduced generation of quantifiable cellular force (up to 70%) over 24 h in both cell types and delayed the onset of measurable contraction (upto sevenfold). The delay of measurable force generation was cell lineage dependent but not FCS dependent. Gene expression of MMP-2, TIMP-2, and collagen type III expression in HDFs were significantly upregulated in constructs of increased stiffness. HNFFs did not show any significant changes in these gene expressions indicating a lineage specific response.
    背景与目标: : 细胞水平的机械和3D空间线索对于在生长,修复或生物反应器中形成的新组织的组织和结构至关重要。成纤维细胞种子的3D胶原蛋白构建体已被用作生物人工细胞外基质 (ECM),为嵌入的常驻细胞提供3D环境。当细胞附着在支架原纤维上时,它们会产生可量化的收缩力,这取决于细胞类型,细胞附着,细胞密度,生长因子和基质刚度。这项研究的目的是量化接种在硬度增加的构建体中的人真皮 (HDF) 和新生儿包皮成纤维细胞 (HNFF) 的细胞机械和分子反应。我们还测试了使用血清饥饿阻断早期附着对这些输出的影响。在凝胶收缩之前,将构建体置于0-10% 的单轴应变下以增加支架刚度,并使用张力培养力监测器 (t-cfm) 监测力的产生。在两种细胞类型中,增加的基质刚度在24小时内减少了可量化的细胞力 (高达70%) 的产生,并延迟了可测量的收缩的开始 (高达7倍)。可测量的力产生的延迟与细胞谱系有关,但与FCS无关。在刚度增加的构建体中,HDFs中MMP-2,TIMP-2和III型胶原表达的基因表达显着上调。HNFFs在这些基因表达中未显示任何显着变化,表明谱系特异性反应。
  • 【A组链球菌胶原蛋白样蛋白1 Scl1通过靶向受伤组织中表达的含细胞纤连蛋白的额外结构域A变体来介导生物膜的形成。】 复制标题 收藏 收藏
    DOI:10.1111/mmi.12125 复制DOI
    作者列表:Oliver-Kozup H,Martin KH,Schwegler-Berry D,Green BJ,Betts C,Shinde AV,Van De Water L,Lukomski S
    BACKGROUND & AIMS: :Wounds are known to serve as portals of entry for group A Streptococcus (GAS). Subsequent tissue colonization is mediated by interactions between GAS surface proteins and host extracellular matrix components. We recently reported that the streptococcal collagen-like protein-1, Scl1, selectively binds the cellular form of fibronectin (cFn) and also contributes to GAS biofilm formation on abiotic surfaces. One structural feature of cFn, which is predominantly expressed in response to tissue injury, is the presence of a spliced variant containing extra domain A (EDA/EIIIA). We now report that GAS biofilm formation is mediated by the Scl1 interaction with EDA-containing cFn. Recombinant Scl1 proteins that bound cFn also bound recombinant EDA within the C-C' loop region recognized by the α(9)β(1) integrin. The extracellular 2-D matrix derived from human dermal fibroblasts supports GAS adherence and biofilm formation. Altogether, this work identifies and characterizes a novel molecular mechanism by which GAS utilizes Scl1 to specifically target an extracellular matrix component that is predominantly expressed at the site of injury in order to secure host tissue colonization.
    背景与目标: : 已知伤口是A组链球菌 (GAS) 的入口。随后的组织定植是由气体表面蛋白和宿主细胞外基质成分之间的相互作用介导的。我们最近报道了链球菌胶原蛋白样蛋白1 Scl1选择性地结合纤连蛋白 (cFn) 的细胞形式,并且还有助于在非生物表面上形成气体生物膜。cFn的一个结构特征主要是对组织损伤的反应,是存在包含额外结构域a (EDA/EIIIA) 的剪接变体。我们现在报告气体生物膜的形成是由Scl1与含EDA的cFn相互作用介导的。结合cFn的重组Scl1蛋白也结合了由 α(9)β(1) 整联蛋白识别的c-c' 环区域内的重组EDA。源自人类真皮成纤维细胞的细胞外2-D基质支持气体粘附和生物膜形成。总之,这项工作确定并表征了一种新的分子机制,通过该机制,GAS利用Scl1特异性靶向主要在损伤部位表达的细胞外基质成分,以确保宿主组织定植。
  • 【抑制Src激酶可阻断高葡萄糖诱导的肾小球系膜细胞EGFR反式激活和胶原合成,并预防小鼠糖尿病肾病。】 复制标题 收藏 收藏
    DOI:10.2337/db12-1010 复制DOI
    作者列表:Taniguchi K,Xia L,Goldberg HJ,Lee KW,Shah A,Stavar L,Masson EA,Momen A,Shikatani EA,John R,Husain M,Fantus IG
    BACKGROUND & AIMS: :Chronic exposure to high glucose leads to diabetic nephropathy characterized by increased mesangial matrix protein (e.g., collagen) accumulation. Altered cell signaling and gene expression accompanied by oxidative stress have been documented. The contribution of the tyrosine kinase, c-Src (Src), which is sensitive to oxidative stress, was examined. Cultured rat mesangial cells were exposed to high glucose (25 mmol/L) in the presence and absence of Src inhibitors (PP2, SU6656), Src small interfering RNA (siRNA), and the tumor necrosis factor-α-converting enzyme (TACE) inhibitor, TAPI-2. Src was investigated in vivo by administration of PP2 to streptozotocin (STZ)-induced diabetic DBA2/J mice. High glucose stimulated Src, TACE, epidermal growth factor receptor (EGFR), mitogen-activated protein kinases (MAPKs), extracellular signal-regulated kinase (ERK1/2, p38), and collagen IV accumulation in mesangial cells. PP2 and SU6656 blocked high glucose-stimulated phosphorylation of Src Tyr-416, EGFR, and MAPKs. These inhibitors and Src knockdown by siRNA, as well as TAPI-2, also abrogated high glucose-induced phosphorylation of these targets and collagen IV accumulation. In STZ-diabetic mice, albuminuria, increased Src pTyr-416, TACE activation, ERK and EGFR phosphorylation, glomerular collagen accumulation, and podocyte loss were inhibited by PP2. These data indicate a role for Src in a high glucose-Src-TACE-heparin-binding epidermal growth factor-EGFR-MAPK-signaling pathway to collagen accumulation. Thus, Src may provide a novel therapeutic target for diabetic nephropathy.
    背景与目标: : 长期暴露于高糖会导致糖尿病肾病,其特征是系膜基质蛋白 (例如胶原蛋白) 积累增加。已经记录了伴随氧化应激的细胞信号和基因表达的改变。检查了对氧化应激敏感的酪氨酸激酶c-Src (Src) 的贡献。在存在和不存在Src抑制剂 (PP2,SU6656),Src小干扰RNA (siRNA) 和肿瘤坏死因子-α 转化酶 (TACE) 的情况下,将培养的大鼠肾小球系膜细胞暴露于高糖 (25 mmol/L) TAPI-2抑制剂。通过向链脲佐菌素 (STZ) 诱导的糖尿病DBA2/J小鼠施用PP2在体内研究了Src。高糖刺激Src,TACE,表皮生长因子受体 (EGFR),丝裂原活化蛋白激酶 (MAPKs),细胞外信号调节激酶 (ERK1/2,p38) 和胶原IV在系膜细胞中的积累。PP2和SU6656阻断了高糖刺激的Src Tyr-416、EGFR和MAPKs的磷酸化。这些抑制剂和siRNA的Src敲除以及TAPI-2也消除了高糖诱导的这些靶标的磷酸化和胶原IV的积累。在STZ糖尿病小鼠中,pp2抑制了蛋白尿,Src pTyr-416增加,TACE激活,ERK和EGFR磷酸化,肾小球胶原积累和足细胞丢失。这些数据表明Src在高葡萄糖-Src-TACE-肝素结合表皮生长因子-egfr-mapk信号通路中与胶原蛋白积累的作用。因此,Src可能为糖尿病肾病提供新的治疗靶标。
  • 【胶原基质在肿瘤血管生成和多形性胶质母细胞瘤进展中的作用。】 复制标题 收藏 收藏
    DOI:10.1016/j.ajpath.2013.06.026 复制DOI
    作者列表:Mammoto T,Jiang A,Jiang E,Panigrahy D,Kieran MW,Mammoto A
    BACKGROUND & AIMS: :Glioblastoma is a highly vascularized brain tumor, and antiangiogenic therapy improves its progression-free survival. However, current antiangiogenic therapy induces serious adverse effects including neuronal cytotoxicity and tumor invasiveness and resistance to therapy. Although it has been suggested that the physical microenvironment has a key role in tumor angiogenesis and progression, the mechanism by which physical properties of extracellular matrix control tumor angiogenesis and glioblastoma progression is not completely understood. Herein we show that physical compaction (the process in which cells gather and pack together and cause associated changes in cell shape and size) of human glioblastoma cell lines U87MG, U251, and LN229 induces expression of collagen types IV and VI and the collagen crosslinking enzyme lysyl oxidase and up-regulates in vitro expression of the angiogenic factor vascular endothelial growth factor. The lysyl oxidase inhibitor β-aminopropionitrile disrupts collagen structure in the tumor and inhibits tumor angiogenesis and glioblastoma multiforme growth in a mouse orthotopic brain tumor model. Similarly, d-penicillamine, which inhibits lysyl oxidase enzymatic activity by depleting intracerebral copper, also exhibits antiangiogenic effects on brain tumor growth in mice. These findings suggest that tumor microenvironment controlled by collagen structure is important in tumor angiogenesis and brain tumor progression.
    背景与目标: : 胶质母细胞瘤是一种高度血管化的脑肿瘤,抗血管生成治疗可改善其无进展生存期。然而,当前的抗血管生成疗法会引起严重的不良反应,包括神经元细胞毒性,肿瘤侵袭性和对治疗的抵抗力。尽管已经提出物理微环境在肿瘤血管生成和进展中起关键作用,但细胞外基质的物理特性控制肿瘤血管生成和胶质母细胞瘤进展的机制尚不完全清楚。在本文中,我们显示了人胶质母细胞瘤细胞系U87MG,U251的物理压实 (细胞聚集并聚集在一起并引起细胞形状和大小相关变化的过程),LN229诱导 ⅳ 型和 ⅵ 型胶原和胶原交联酶赖氨酰氧化酶的表达,并上调血管生成因子血管内皮生长因子的体外表达。在小鼠原位脑肿瘤模型中,赖氨酰氧化酶抑制剂 β-氨基丙腈破坏肿瘤中的胶原蛋白结构并抑制肿瘤血管生成和多形性胶质母细胞瘤的生长。同样,d-青霉胺通过消耗脑内铜来抑制赖氨酰氧化酶的酶活性,也对小鼠脑肿瘤生长表现出抗血管生成作用。这些发现表明,胶原结构控制的肿瘤微环境在肿瘤血管生成和脑肿瘤进展中很重要。
  • 【用于压力性尿失禁的非手术,射频胶原蛋白变性: 回顾性3年评估。】 复制标题 收藏 收藏
    DOI:10.1586/17434440.4.4.455 复制DOI
    作者列表:Appell RA,Singh G,Klimberg IW,Graham C,Juma S,Wells WG,Kanellos A,Reilley SF
    BACKGROUND & AIMS: :Transurethral radiofrequency collagen denaturation, a nonsurgical treatment for stress urinary incontinence, reduces regional dynamic tissue compliance without causing tissue necrosis or gross tissue shrinkage, unlike transvaginal radiofrequency tissue ablation. This retrospective study evaluated long-term safety and efficacy in 21 patients from a 12-month, randomized controlled trial utilizing 3-day diaries and the Incontinence Quality of Life (I-QOL) survey. Significant increases in overall I-QOL scores 3 years or more post treatment was the primary end point. Secondary end points were reductions in frequency and severity of incontinence episodes. After 3 years, mean overall I-QOL score improvement was 12.7 (+/-26); 56% of patients achieved 50% or more reduction in frequency. No new adverse events occurred. These results indicate that radiofrequency collagen denaturation is safe and provides durable efficacy.
    背景与目标: : 与经阴道射频组织消融不同,经尿道射频胶原变性是一种非手术治疗压力性尿失禁的方法,可降低局部动态组织顺应性,而不会引起组织坏死或大组织收缩。这项回顾性研究评估了一项为期12个月的随机对照试验中21例患者的长期安全性和有效性,该试验利用3天日记和尿失禁生活质量 (I-QOL) 调查。治疗后3年或更长时间的总体I-QOL评分显着增加是主要终点。次要终点是尿失禁发作频率和严重程度的降低。3年后,平均I-QOL评分改善12.7 (+/-26); 56% 的患者达到50% 或更多的频率降低。没有新的不良事件发生。这些结果表明,射频胶原蛋白变性是安全的,并提供持久的功效。
  • 【使用比率非线性光学显微镜对癌性食管组织中的胶原纤维进行无标记表征。】 复制标题 收藏 收藏
    DOI:10.1177/1535370220934039 复制DOI
    作者列表:Chen WC,Chen YJ,Lin ST,Hung WH,Chan MC,Wu IC,Wu MT,Kuo CT,Das S,Kao FJ,Zhuo GY
    BACKGROUND & AIMS: IMPACT STATEMENT:The issue of classifying esophageal cancer at various developmental stages is crucial for determining the optimized treatment protocol for the patients, as well as the prognosis. Precision improvement in staging esophageal cancer keeps seeking quantitative and analytical imaging methods that could augment histopathological techniques. In this work, we used nonlinear optical microscopy for ratiometric analysis on the intrinsic signal of two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) from single collagen fibers only in submucosa of esophageal squamous cell carcinoma (ESCC). The blind tests of TPEF/SHG and forward (F)/backward (B) SHG were demonstrated to compare with the histology conclusion. The discussion of sensitivity and specificity was provided via statistical comparison between the four stages of esophageal cancer. To the best of our knowledge, this is the first study of using these two ratios in combination for staging ESCC.
    背景与目标: 影响声明: 在各个发育阶段对食管癌进行分类对于确定患者的最佳治疗方案以及预后至关重要。食管癌分期的精度提高一直在寻求可以增强组织病理学技术的定量和分析成像方法。在这项工作中,我们使用非线性光学显微镜对食管鳞状细胞癌 (ESCC) 粘膜下层的双光子激发荧光 (TPEF) 和二次谐波产生 (SHG) 的固有信号进行了比率分析。TPEF/SHG和向前 (F)/向后 (B) SHG的盲测试被证明与组织学结论进行了比较。通过统计比较四个阶段的食管癌之间的敏感性和特异性的讨论。据我们所知,这是首次将这两种比率组合用于ESCC分期的研究。

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