自由基与软骨基质胶原蛋白类型改变

本文利用ＳＤＳ－聚丙烯酰胶电泳方法,定量研究了体外培养的软骨细胞和软骨组织基质中Ⅱ型胶原蛋白的含量。结果表明氧自由基（．Ｏ２＾－和．ＯＨ）和具有自由基性质的物质（黄腐酸,镰刀菌毒素）可使软骨细胞合成,分泌异常的非Ⅱ型的胶原蛋白,同时,硒化合物可明显地抑制此种效应。     

胶原蛋白与Ⅵ型胶原蛋白的结构概述

描述了胶原蛋白的结构和Ⅵ型胶原蛋白的结构以及胶原蛋白的应用 ,并对人类胶原蛋白的生产进行了展望。     

力学环境对软骨基质代谢的影响

正常关节软骨所受压力是由动态压力与静态压力交替完成。压力引起软骨一系列生理变化包括细胞及细胞外基质成分变形、组织内液体流动、水流电位和生理生化变化。这些变化直接调控细胞外基质代谢。体外构建有良好功能的组织工程化软骨是目前软骨病变、缺损理想的修复方法。研究力学环境对软骨基质代谢的影响,对构建组织工程化软骨有深远意义。     

胶原蛋白的电子活性与矽肺纤维化Ⅱ正常大鼠与实验性矽肺大鼠肺Ⅰ型胶原稳定自由基的研究

由正常大鼠肺撮取的酸溶性Ⅰ型胶原出现一个明显的ESR信号(g=2.006),而由矽肺大鼠提取的酸溶性Ⅰ型胶原较少或缺乏该ESR信号,该信号不是顺磁过渡元素产生,而是稳定自由基的反映.肺胶原可能存在两种稳定的自由基,一种易受紫外线辐照的激发并且在电场极化下自旋耦合而减弱,另一种自由基则相对稳定,对紫外线辐照和电场极化不敏感.矽肺胶原缺乏活泼性较高的稳定自由基.     

胶原蛋白的开发与应用研究进展

胶原蛋白(collagen)是一类哺乳动物细胞外基质中的主要结构蛋白,广泛地存在于皮肤、骨骼、肌肉等组织中,主要参与细胞的增殖、分化、迁移和信号传递等生理生化行为,对组织细胞等起着支撑、修复、保护的作用。由于胶原蛋白具有良好的生物学特性,其在组织工程、临床医学、食品工业、包装材料、化妆品、医学美容、生物材料以及医疗器械等方面都有着广泛的应用。本文综述了胶原蛋白的生物学特性及其在国内外生物工程研究开发中的研究进展,并对胶原蛋白未来开发的前景进行了展望。     

红景天苷对运动后自由基和能量代谢改变的影响

目的:探讨红景天苷在运动过程中对自由基和能量代谢相关指标的作用,从抗氧化系统、能量代谢系统等方面研究红景天苷抗运动性疲劳的机制。方法:本研究采用小鼠运动模型,40只雄性小鼠随机取分为4组(n=10):红景天苷运动组,红景天苷安静组,运动对照组,安静对照组。红景天苷运动组和安静组两组以150 mg/(kg.d)的红景天苷灌胃给药,运动和安静对照组两组以同样体积蒸馏水灌胃,连续给药2周;末次灌胃30 min后,运动组进行无负重游泳120 min,然后测定与运动性疲劳相关的生化指标。结果:研究结果表明,红景天苷能够提高运动小鼠肝脏内超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)等抗氧化酶活性,降低丙二醛(MDA)含量;红景天苷具有稳定运动小鼠血糖,增加肝、肌糖原储备,防止长时间运动后血糖和肝、肌糖原水平降低的作用;红景天苷可提高运动小鼠血浆总胆固醇(TC)、甘油三酯(TG)及游离脂肪酸(FFA)的水平,有影响不同状态下的脂肪代谢,促进脂肪利用的作用。结论:红景天苷对运动过程中自由基和能量代谢改变的影响是其抗运动性疲劳的机制之一。     

Effect of Oxygen Free Radicals on Corneal Collagen

Corneal collagen was labeled in vivo by injection of ¹⁴C-proline into the anterior chamber of rabbit eyes. The isolated corneal collagen was incubated in iron-free phosphate buffered saline (pH 7.4) containing I mM axorbate and 0.1 mM CuSO₄ for either 1 hour or 3 hours at 37°. Addition of 2 volumes of 8 M urea-I mM dithiothreitol and heating for 1 min at 100° solubilized virtually all of the collagen in the control incubations but left a significant amount of insoluble collagen in specimens exposed to the hydroxyl radical generating system. This residue amounted to 19% and 38% of the initial radioactivity in samples incubated for 1 h and 3 h, respectively. The chromatographic profiles (gel filtration on CL-4B) of the soluble fraction showed an increase in both aggregation and degradation products of collagen in the 1 h incubation mixture, whereas after 3 h there was an increase only in degradation products. These observations suggest that additional crosslinking of the soluble collagen aggregates observed at 1 h may be responsible for their subsequent disappearance at 3 h, with concomitant increase of the insoluble fraction. Collagen degradation by OH may play a role in corneal ulceration, whereas hydroxyl radical-mediated crosslinking is consistent with age-dependent increases in insoluble collagen.     

Effect of Oxygen Free Radicals on Corneal Collagen

Corneal collagen was labeled in vivo by injection of ¹⁴C-proline into the anterior chamber of rabbit eyes. The isolated corneal collagen was incubated in iron-free phosphate buffered saline (pH 7.4) containing I mM axorbate and 0.1 mM CuSO₄ for either 1 hour or 3 hours at 37°. Addition of 2 volumes of 8 M urea-I mM dithiothreitol and heating for 1 min at 100° solubilized virtually all of the collagen in the control incubations but left a significant amount of insoluble collagen in specimens exposed to the hydroxyl radical generating system. This residue amounted to 19% and 38% of the initial radioactivity in samples incubated for 1 h and 3 h, respectively. The chromatographic profiles (gel filtration on CL-4B) of the soluble fraction showed an increase in both aggregation and degradation products of collagen in the 1 h incubation mixture, whereas after 3 h there was an increase only in degradation products. These observations suggest that additional crosslinking of the soluble collagen aggregates observed at 1 h may be responsible for their subsequent disappearance at 3 h, with concomitant increase of the insoluble fraction. Collagen degradation by OH may play a role in corneal ulceration, whereas hydroxyl radical-mediated crosslinking is consistent with age-dependent increases in insoluble collagen.     

Type III Collagen,a Fibril Network Modifier in Articular Cartilage

The collagen framework of hyaline cartilages, including articular cartilage, consists largely of type II collagen that matures from a cross-linked heteropolymeric fibril template of types II, IX, and XI collagens. In the articular cartilages of adult joints, type III collagen makes an appearance in varying amounts superimposed on the original collagen fibril network. In a study to understand better the structural role of type III collagen in cartilage, we find that type III collagen molecules with unprocessed N-propeptides are present in the extracellular matrix of adult human and bovine articular cartilages as covalently cross-linked polymers extensively cross-linked to type II collagen. Cross-link analyses revealed that telopeptides from both N and C termini of type III collagen were linked in the tissue to helical cross-linking sites in type II collagen. Reciprocally, telopeptides from type II collagen were recovered cross-linked to helical sites in type III collagen. Cross-linked peptides were also identified in which a trifunctional pyridinoline linked both an α1(II) and an α1(III) telopeptide to the α1(III) helix. This can only have arisen from a cross-link between three different collagen molecules, types II and III in register staggered by 4D from another type III molecule. Type III collagen is known to be prominent at sites of healing and repair in skin and other tissues. The present findings emphasize the role of type III collagen, which is synthesized in mature articular cartilage, as a covalent modifier that may add cohesion to a weakened, existing collagen type II fibril network as part of a chondrocyte healing response to matrix damage.     

运动对人体自由基代谢的影响

自由基代谢普遍存在于机体各组织。正常情况下,人体自由基的产生与清除处于平衡状态。而人体自由基产生过多或机体清除自由基能力下降将给机体带来损伤。本文采用文献资料法,阐述了自由基的类型、功能、自由基产生的机制以及抗氧化系统的种类等基本理论。通过对力竭运动、耐力运动和无氧运动3种不同形式的运动对自由基代谢及抗氧化酶活性的研究得出结论:不同的运动形式及负荷方式对体内自由基代谢及抗氧化酶活性将产生不同影响;运动对自由基代谢影响具有器官与组织差异性。这些研究成果为科学指导运动训练和健身活动、快速有效的消除运动性疲劳并增强运动能力提供重要的科学理论依据。     

Free Radicals in the Atmosphere

Like the oxidation in a flame, the oxidation in the atmosphere is mediated by free radicals. Unlike a flame, however, atmospheric oxidation needs an external source of energy: the sun light. In fact the most important radical acting in the lower atmosphere, the hydroxyl radical, OH, is produced following the UV-photolysis of ozone, O,which yields an excited oxygen atom, O'D:

OH reacts with most atmospheric trace gases, in many cases as the first and rate determining step in the reaction chain leading to oxidation. In this way a host of various other radicals (e.g. peroxy radicals), most of them very short lived, are generated. Usually these oxidation reactions form chains which regenerate OH, thus maintaining OH at a relatively high concentration level on the order of 10⁶cm∼³ during the day. The reactions which control the OH concentration will be discussed in detail. During the night radical formation is greatly diminished. It proceeds, for example, through the reaction of defines with O, and. in dry air, through reaction of defines and aldehydes with the nitrate radical, NO,.     

Free Radicals in the Atmosphere

Like the oxidation in a flame, the oxidation in the atmosphere is mediated by free radicals. Unlike a flame, however, atmospheric oxidation needs an external source of energy: the sun light. In fact the most important radical acting in the lower atmosphere, the hydroxyl radical, OH, is produced following the UV-photolysis of ozone, O,which yields an excited oxygen atom, O'D:

OH reacts with most atmospheric trace gases, in many cases as the first and rate determining step in the reaction chain leading to oxidation. In this way a host of various other radicals (e.g. peroxy radicals), most of them very short lived, are generated. Usually these oxidation reactions form chains which regenerate OH, thus maintaining OH at a relatively high concentration level on the order of 10⁶cm～³ during the day. The reactions which control the OH concentration will be discussed in detail. During the night radical formation is greatly diminished. It proceeds, for example, through the reaction of defines with O, and. in dry air, through reaction of defines and aldehydes with the nitrate radical, NO,.     

Type IX Collagen Interacts with Fibronectin Providing an Important Molecular Bridge in Articular Cartilage

Type IX collagen is covalently bound to the surface of type II collagen fibrils within the cartilage extracellular matrix. The N-terminal, globular noncollagenous domain (NC4) of the α1(IX) chain protrudes away from the surface of the fibrils into the surrounding matrix and is available for molecular interactions. To define these interactions, we used the NC4 domain in a yeast two-hybrid screen of a human chondrocyte cDNA library. 73% of the interacting clones encoded fibronectin. The interaction was confirmed using in vitro immunoprecipitation and was further characterized by surface plasmon resonance. Using whole and pepsin-derived preparations of type IX collagen, the interaction was shown to be specific for the NC4 domain with no interaction with the triple helical collagenous domains. The interaction was shown to be of high affinity with nanomolar K_d values. Analysis of the fibronectin-interacting clones indicates that the constant domain is the likely site of interaction. Type IX collagen and fibronectin were shown to co-localize in cartilage. This novel interaction between the NC4 domain of type IX collagen and fibronectin may represent an in vivo interaction in cartilage that could contribute to the matrix integrity of the tissue.     

自由基和基质金属蛋白酶介导脑缺血血脑屏障损伤的研究进展

血脑屏障的破坏是引起脑缺血损伤及继发水肿、出血、炎症的微观原因。缺血缺氧和再灌注过程产生的自由基,以及后续基质金属蛋白酶的激活,是破坏血脑屏障结构和功能的重要分子机制。因而,在脑缺血早期及时抑制自由基产生并清除自由基,抑制基质金属蛋白酶的活性,是降低脑缺血血脑屏障损伤及其并发症的关键环节。本文将从血脑屏障损伤的角度,概述自由基与基质金属蛋白酶在脑缺血损伤过程中的作用。     

Free Radicals in Melanin-Cationic Porphyrins Complexes in the Dark and Under Light Irradiation

Electron Paramagnetic Resonance (EPR) spectroscopy was employed in the study of the interaction between L-3,4-dihydroxyphenylalanine (L-Dopa) melanin and the cat-ionic porphyrins meso-tetrakis(1-methylpyridinium-4yl)-porphyrin (TMPyP), meso-tetrakis-(1-benzylpyridinium-4-yl)-porphyrin (TBzPyP), and their respectives complexes ZnTMPyP and ZnTBzPyP. By monitoring signal intensities and progressive microwave power saturation it was shown that the interaction increases the equilibrium concentration of free radicals in L-Dopa melanin in the dark. The extent of increase is dependent on the presence of molecular oxygen and on the type of porphyrin. Not all interacting sites available for complexation in L-Dopa melanin are involved in the formation of free radicals. It was also observed that the interaction with porphyrins promotes an increase in the number of photoinduced free radicals in L-Dopa melanin during illumination with visible light.     

螺旋藻转化纳米元素硒的制备及其体外清除自由基活性的初步研究

研究利用高密度富硒螺旋藻(Se-SP)细胞通过生物转化制备纳米元素硒(Nano-Se)的可行性,观察Nano-Se在体外对氧自由基的清除作用。用梯度离心分选Nano-Se,原子力显微镜(AFM)、透射电镜(TEM)及X-射线能谱(EDX)联用表征纳米粒中的元素硒形态,电感耦合等离子质谱仪(ICP-MS)测定Nano-Se中的硒含量,化学发光方法检测Nano-Se在体外对超氧自由基和羟自由基的清除作用。结果发现,Nano-Se主要由元素硒构成,形态呈球形,73%的纳米粒子直径大小分布在(61±17)nm范围。Nano-Se在体外对两种氧自由基的最大清除率分别为:30.1%和27.6%,相应的EC50分别为:0.8 μg/ml和2.2 μg/ml。相同剂量时,Nano-Se对氧自由基的清除作用比硒代蛋氨酸(Se-Met)及Se-SP中其它含硒活性成分更强。结果提示,利用高密度Se-SP可诱导Nano-Se的大量生成,Se-SP转化的Nano-Se可能是一种新的抗氧化硒形态,其作用机制和体内生物活性有待深入研究。