Treatment with chondroitinase ABC alleviates bleomycin-induced pulmonary fibrosis

Pulmonary fibrosis is characterized by an accumulation of inflammatory cells in the lung interstitium, followed by an increased deposition of extracellular matrix. Macrophages play a vital role in this disease by mediating the progression from inflammation to fibrosis, but the mechanisms by which macrophages are retained at these sites are not fully understood. Although the transmigration of leukocytes is regulated by chemokines, glycosaminoglycans modulate the function of chemokines and the migration of leukocytes. Accordingly, we investigated the role of chondroitin sulfate proteoglycans (CSPGs) in a murine bleomycin-induced pulmonary fibrosis models. After intratracheal injection of bleomycin or saline, mice were randomized to receive one intravenous injection and continuous infusion of the CSPG-digesting enzyme chondroitinase ABC (ChABC), or vehicle, for 7 days. CSPGs were readily induced and progressively augmented after the bleomycin challenge. Although CSPGs inhibited the early CCL2-dependent recruitment of macrophages, deposited CSPGs retained macrophages in fibrotic interstitium in a CD44-dependent manner. Treatment with ChABC in vivo dramatically increased survival of the mice and reduced collagen deposition by inhibiting persistent macrophage accumulation. These results indicate a pivotal role for CSPGs in macrophage-mediated lung fibrogenesis and suggest a possible new therapeutic role for ChABC in pulmonary fibrosis.     

Degradation of cartilage proteoglycans by elastase is dependent on charge-mediated interactions

Summary We investigated the mechanism of cartilage degradation by pancreatic elastase as a model system for the action of cationic proteases such as leukocyte elastase and cathepsin-G. It is shown that the cationic properties of elastase contribute to its proteolytic potential with respect to cartilage degradation. Elastase preparations with neutral or negative charge, obtained by chemical modification were far less effective in cartilage degradation than the cationic, native enzyme. Modification of elastase did not affect the active site of the enzyme as shown by kinetic studies, nor did it alter the complexing with alpha-1-proteinase inhibitor. Quantitative and autoradiographic studies indicate that the positive charge of the enzyme favors the penetration in cartilage and interaction with the polyanionic proteoglycan substrate. It is concluded that the potent cartilage-degrading activity of elastase is dependent on charge-mediated interactions with its substrate.     

实验性蛋白多糖降解致兔膝关节软骨早期退变的核磁共振成像研究

目的　探讨低场磁共振成像 (MRI)在检测骨关节炎 (OA)早期软骨退变的表现及其价值。方法　选用新西兰大白兔 32只 ,右侧膝关节腔内注射 0 1ml木瓜蛋白酶溶液 (5U) ,建立OA早期软骨退变的动物模型。并在注药前及注药后 2 4、4 8、72h行双侧膝关节矢状面GE准T2 WI、SE T1WI、SE PDWI、SE T2 WI序列成像 ,取关节软骨组织作蛋白多糖含量测定和组织病理学检查。结果　注射木瓜蛋白酶后 2 4、4 8h ,关节软骨明显变薄 ,磁共振 (MR)信号强度明显降低。与对照组比较 ,两者差异均有显著性 (P <0 0 5 )。至注药后 72h关节软骨的厚度及信号强度已基本恢复正常。与对照组比较 ,两者差异无显著性 (P >0 0 5 )。关节软骨蛋白多糖含量测定及组织学检查结果表明 ,注射木瓜蛋白酶后 2 4、4 8h ,蛋白多糖含量明显降低 ,至注药后 72h ,蛋白多糖含量已逐渐恢复。软骨细胞均未见异常改变。结论　通过MR检查 ,可发现早期的软骨退变。     

Thiazolidinediones reduce the LDL binding affinity of non-human primate vascular cell proteoglycans

Aims/hypothesis Retention of atherogenic lipoproteins in the artery wall by proteoglycans is a key step in the development of atherosclerosis. Thiazolidinediones have been shown to reduce atherosclerosis in mouse models. The aim of this study was to determine whether thiazolidinediones modify vascular proteoglycan synthesis in a way that decreases LDL binding.Methods Primate aortic smooth muscle cells were exposed to troglitazone or rosiglitazone, or no stimulus at all for a 24-hour steady-state labelling period. Sulphate incorporation, size and LDL binding affinity of proteoglycans were determined. Proteoglycans secreted by cells in the presence or absence of troglitazone were separated into large and small classes by size exclusion chromatography, and LDL binding affinity was determined.Results Proteoglycans synthesised by cells exposed to troglitazone or rosiglitazone were smaller, with decreased sulphate incorporation and decreased LDL binding affinity. However, troglitazone had a greater effect than rosiglitazone. Troglitazone reduced the LDL binding affinities of both the large and small proteoglycans compared with control. The binding differences persisted when glycosaminoglycan chains released from proteoglycans were incubated with LDL, indicating that troglitazone affects the glycosaminoglycan synthetic machinery of these cells.Conclusions/interpretation Thiazolidinediones decrease the LDL binding affinity of the proteoglycans synthesised by primate aortic smooth muscle cells. This could, in part, account for the reduced atherosclerosis observed in animal models.Abbreviations PPAR peroxisome proliferator-activated receptor - K_d binding constantPresented in part at the 3rd Annual Conference on Arteriosclerosis, Thrombosis and Vascular Biology, Salt Lake City, Utah, USA, 6 April 2002     

饰胶蛋白聚糖对大鼠肾系膜细胞生长的抑制作用及其信号转导途径的研究

目的 探讨饰胶蛋白聚糖(DCN)对大鼠系膜细胞(MsC)生长的抑制作用及其信号转导分子MAPKs和p21表达的影响&#65377;方法 经脂质体介导将DCN基因转染体外培养的大鼠MsC,筛选和鉴定后,收集阳性细胞克隆的培养上清(DCN上清),将其加入正常MsC的培养液中, 采用流式细胞仪检测细胞周期&#65377;用Western 印迹法分别检测MAPKs,包括细胞外调节激酶(ERK)1/2&#65380;应激活化蛋白激酶(SAPK)/氨基末端激酶(JNK)和p38和p21蛋白表达;用免疫荧光法观察p21在细胞中的表达&#65377;结果 DCN上清明显抑制正常MsC的增殖, G2-M期细胞数明显减少,仅为对照组的35%(P < 0.05); 磷酸化ERK1/2及SAPK/JNK表达增强, 分别为对照组的2.2倍&#65380; 1.4倍及1.7倍&#65380; 1.8倍;磷酸化p38无明显变化&#65377;DCN抗体呈浓度依赖性抑制磷酸化ERK1/2&#65380; SAPK/JNK的表达上调&#65377;DCN上清还可使细胞p21的表达明显增强,而DCN抗体也同样呈浓度依赖性地抑制其上调表达的作用, ERK1/2 及SAPK/JNK通路抑制剂U0126和circumin均可抑制其上调表达的作用,分别为对照组的64%和61%;而p38通路抑制剂SB203580则对其无影响&#65377;结论 DCN对肾MsC的生长有抑制作用,其机制可能经ERK1/2&#65380;SAPK/JNK和p21蛋白介导&#65377;     

Structure and biological activity of the extracellular matrix

 The extracellular matrix is formed by complex and intricate networks within which molecules are precisely organized. These molecular networks determine the specific histoarchitecture of tissues and provide cells with information and a scaffold. Most of the structural extracellular matrix molecules – collagens, noncollagenous glycoproteins, and proteoglycans – are chimeric and share common domains. Studies of the interactions between extracellular matrix molecules and mapping of the interaction sites to defined structural modules have led to the concept that the function of the extracellular matrix relies largely in the polymers that they form. Furthermore, determination of the tertiary structure of protein motifs involved either in the assembly of the various molecules into polymers or in cell–extracellular matrix interactions has recently opened the field of structural biology of the extracellular matrix. Received: 17 April 1997 / Accepted: 24 September 1997     

Resistant proteoglycans in epiphyseal plate cartilage. Variations in their distribution in relationship to age and species

The localizations of resistant proteoglycans (RPGs) in the epiphyseal plates of rats, dogs, and humans are similar. In the epiphyseal plates from young rats, dogs, and humans, the RPGs form a stratum at the junction of the zones of resting and proliferating cells. Non-calcified cartilage RPGs are associated with cells which have the potential for proliferation or column organization. As the individuals age, RPGs are found in intercolumnar regions or at times are even absent. There is also a type of RPGs in calcified cartilage, including the calcified cartilage subjacent to the articular surface, in all species. In human epiphyseal plates, looser fibrillar RPGs change abruptly to a more condensed type in the zone of provisional calcification. Calcified cartilage RPGs stain more intensely with toluidine blue and may represent a different type of RPGs.     

Collagen hydrolysate for the treatment of osteoarthritis and other joint disorders:a review of the literature

ABSTRACT

Background: There is a need for an effective treatment for the millions of people in the United States with osteoarthritis (OA), a degenerative joint disease. The demand for treatments, both traditional and non-traditional, will continue to grow as the population ages.

Scope: This article reviews the medical literature on the preclinical and clinical research on a unique compound, collagen hydrolysate. Articles were obtained through searches of the PubMed database (www.pubmed.gov) through May 2006 using several pairs of key words (collagen hydrolysate and osteoarthritis; collagen hydrolysate and cartilage; collagen hydrolysate and chondrocytes; collagen hydrolysate and clinical trial) without date limits. In addition, other sources of information, such as abstracts presented at scientific congresses and articles in the German medical literature not available on PubMed, were reviewed and included based on the authors’ judgment of their relevance to the topic of the review.

Findings: According to published research, orally administered collagen hydrolysate has been shown to be absorbed intestinally and to accumulate in cartilage. Collagen hydrolysate ingestion stimulates a statistically significant increase in synthesis of extracellular matrix macromolecules by chondrocytes (?p < 0.05 compared with untreated controls). These findings suggest mechanisms that might help patients affected by joint disorders such as OA. Four open-label and three double-blind studies were identified and reviewed; although many of these studies did not provide key information – such as the statistical significance of the findings – they showed collagen hydrolysate to be safe and to provide improvement in some measures of pain and function in some men and women with OA or other arthritic conditions.

Conclusion: A growing body of evidence provides a rationale for the use of collagen hydrolysate for patients with OA. It is hoped that ongoing and future research will clarify how collagen hydrolysate provides its clinical effects and determine which populations are most appropriate for treatment with this supplement.     

人颈椎椎体终板软骨细胞退变模型的建立及其意义

目的 建立人颈椎椎体终板软骨细胞退变模型,观察人正常颈椎椎体和退变颈椎椎体终板软骨细胞的形态及表征.方法 选择2010年7月至2011年7月49例颈椎骨折、脱位(19例)及颈椎病(30例)患者术中取出的颈椎终板软骨,用酶消化法分别分离培养人正常颈椎椎体终板软骨细胞(对照组)和退变颈椎椎体终板软骨细胞(颈椎病组);用倒置显微镜和HE染色法观察细胞形态学变化;四甲基偶氮唑蓝(MTT)法绘制细胞生长曲线;甲苯蓝染色及反转录-PCR(RT-PCR)法对终板软骨细胞进行鉴定;RT-PCR法检测终板软骨细胞特征性基因蛋白多糖、Ⅱ型胶原及Ⅰ型胶原的表达.结果 人颈椎椎体终板软骨细胞表达特征性蛋白多糖、Ⅱ型胶原及Ⅰ型胶原,其生长情况及细胞表型类似于关节软骨细胞.对照组原代终板软骨细胞以多角形为主,增殖速度较快;而颈椎病组原代终板软骨细胞以梭形为主,细胞增殖速度较慢.颈椎病组原代终板软骨细胞表达的蛋白多糖基因(0.695 ±0.052)和Ⅱ型胶原基因(0.726 ±0.035)均低于对照组(0.950±0.032、0.907±0.078,t=7.263、3.681,P=0.002、0.021),Ⅰ型胶原基因则高于对照组(0.795±0.028比0.552±0.070,t=-5.560,P=0.005).结论 成功建立了人颈椎椎体终板软骨细胞退变模型,为椎间盘退变机制研究提供了较好的细胞学基础,解决了以前一直以动物细胞模型为研究对象的局限性.