关键词： Asporin   Small leucine-rich repeat proteoglycans   Osteoarthritis  

摘要： Objective: To provide an overview of the literature describing the role of asporin, a small leucine-rich proteoglycan (SLRP), in osteoarthritis (OA). Method: A literature search was performed and reviewed using the narrative approach. Results: As a class I SLRP member, asporin, is distinct from other SLRPs. Accumulating evidence demonstrates the involvement of asporin in OA pathogenesis. Many human studies have been conducted to explore the association between the D-repeat polymorphisms and OA susceptibility, but these yield inconsistent results. Possible mechanisms for the involvement of asporin in OA pathology include its influence on TGF-beta (transforming growth factor-beta) signaling pathways and collagen mineralization. To date, no studies were found to use an asporin-deficient animal model that would help to understand disease mechanisms. Many issues must be addressed to clarify the link between asporin and OA to provide a novel therapeutic strategy for OA, perhaps through controlling and modifying the TGF-beta-ECM system. Conclusions: Studies examined demonstrate the involvement of asporin in OA pathogenesis, and possible mechanisms by which asporin may be involved in this process have been proposed. However, large-scale interracial studies should be conducted to investigate the association between asporin and OA, and further investigations are needed to obtain a better understanding of the disease mechanism, develop novel therapeutic strategies, and explore new approaches for diagnosis of OA. (C) 2015 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

关键词： L-Sox5   Chondrogenesis   Runx2   Hypertrophy   Asporin   Osteoarthritis  

摘要： L-Sox5 is a member of sex-determining region Y-type high mobility group box (SOX) family of transcription factors. We assessed the effects of retroviral overexpression of L-Sox5 on chondrocyte differentiation using the clonal murine cell line ATDC5. We observed a temporal-restricted expression pattern of L-Sox5 in insulin-induced ATDC5 cells differentiating toward chondrocyte lineage. The protein expression levels of L-Sox5 showed a drastic decrease in contrast to unaltered mRNA levels during differentiation. L-Sox5 delayed the differentiation of ATDC5 cells as evidenced by Alcian blue staining for proteoglycan synthesis. The mRNA levels of chondrocyte and hypertrophic/osteoarthritic markers were markedly decreased or delayed in L-Sox5 overexpressing cells. L-Sox5 abrogated the promoter activity of Runx2. These results suggest that L-Sox5 protein expression may diminish along with the progress of chondrogenic differentiation. L-Sox5 may act as a negative regulator if expressed aberrantly at least in part by regulating the critical fate of chondrogenesis.

关键词： 骨形态发生蛋白   椎间盘退变   基因治疗  

摘要： 椎间盘退变(intervertebral disc degeneration,IDD)主要见于腰椎间盘突出、椎管狭窄等疾病,其引起的肌肉劳损、腰背疼痛等症状严重困扰患者的正常生活。骨形态发生蛋白(bone morphogenetic proteins,BMPs)已被证实是一类与骨形成及软骨形成具有密切相关性的生长因子,在椎间盘组织的发展形成与功能维持中发挥的效应吸引了大量研究者探索其机理,而通过基因治疗的途径早期防治IDD是目前的研究热点之一,本文就BMPs在IDD修复过程中作用机制的研究进展进行综述。

关键词： aspirin   CAF   rac1   scirrhous gastric cancer   SLRP  

摘要： Scirrhous gastric cancer, which has the worst prognosis among the various types of gastric cancer, is highly invasive and associated with abundant stromal fibroblasts. Although cancer-associated fibroblasts (CAFs) have been proposed to generate a tumor-supportive extracellular matrix that promotes the expansion of this type of cancer, the molecular mechanisms by which CAFs assist cancer cells are not yet fully understood. Here, we show for the first time that Asporin, a small leucine-rich proteoglycan (SLRP), is predominantly expressed in CAFs, and has essential roles in promoting co-invasion of CAFs and cancer cells. CAFs of scirrhous gastric cancer possess high potential for invasion, and invasion by CAFs frequently proceeded invasion by cancer cells, both in vitro and in vivo. Expression of Asporin was induced in fibroblasts by exposure to gastric cancer cells. Asporin secreted from CAFs activates Rac1 via an interaction with CD44 and promotes invasion by CAFs themselves. Moreover, Asporin promoted invasion by neighboring cancer cells, via paracrine effects mediated by activation of the CD44-Rac1 pathway. These results suggest that Asporin is a unique SLRP that promotes progression of scirrhous gastric cancer and is required for coordinated invasion by CAFs and cancer cells. Therefore, Asporin may represent a new therapeutic target molecule for the development of drugs aimed at manipulating the cancer microenvironment.

关键词： 椎间盘退变   信号通路   作用机制   核转录因子   基因治疗   髓核细胞  

摘要： 第一部分:NF-κB信号通路在退变椎间盘中的表达及作用　　研究背景:椎间盘退行性病变(Intervertebral disc degeneration，IVDD)是脊柱退行性病变的前提和基础病理。椎间盘退变所继发的腰椎间盘突出症、腰椎管狭窄症、盘源性腰痛和腰椎不稳症等疾患是腰腿痛的主要病因。椎间盘退变病因众多，机制复杂。临床上表现为发病率高、涉及人群广、易复发，是严重影响人类健康的一大类疾患，目前无有效的治疗措施。针对椎间盘退变所继发的各类疾患的最主要治疗措施是切除退变的椎间盘组织，但是这些措施并未有效地控制椎间盘退变的发病率，也没有降低高额的医疗费用，同时手术带来的各种严重并发症影响患者的身心健康。因此，从分子水平深入研究椎间盘退变的病理生理机制，从根本上延缓椎间盘退变，减轻国民经济负担，改变传统手术治疗，已经成为目前重要的研究方向之一。　　椎间盘退变是一个多因素协同作用、不断进展的慢性过程，国外学者从分子生物学、生物化学、生物力学、免疫学等角度对椎间盘退变的机制进行了研究。尚未发现可强力干预椎间盘退变并可持续发挥作用的方法。大多数国外学者建议通过有效抑制椎间盘纤维化、促进细胞外基质合成、抑制细胞外基质降解，可延缓椎间盘退变的发展。　　核转录因子(NF)-κB(nuclear factor-κ B，NF-κB)可调节多种细胞因子和酶等基因的表达，参与机体多种病理生理过程，如免疫应答、炎症相关分子表达，炎症的发生、细胞增殖、细胞外基质的降解和抗凋亡等。在正常生理情况下，NF-κB以无活性的状态存在于细胞浆中;当细胞受到压力，氧化等外界刺激时，NF-κB迅速转移到核内，发挥转录调控作用。越来越多的证据提示NF-κB的激活是退行性疾病的主要调节枢纽，如骨关节炎、骨质疏松、类风湿关节炎和肌肉营养不良等。在软骨细胞中，NF-κB介导MMP-1（matrix metalloproteinases-1）、MMP-3(matrix metalloproteinases-3)的表达。上述结果提示，NF-κB在椎间盘退变及骨关节炎中起到的重要作用而吸引广泛的关注。尽管，目前有一些文献报道NF-κB在椎间盘退变性疾病中具有重要的意义，然而有关系统性研究NF-κB在椎间盘内的表达及机制尚未有相关研究报道。本实验首次系统性阐述了，NF-κB在椎间盘组织中的表达及作用。　　目的:研究NF-κB信号通路在退变椎间盘中的表达及意义，探讨NF-κB信号通路在椎间盘退变中的作用，为下一步治疗椎间盘退变的药物及基因治疗开发，提供相应的理论依据。　　方法:通过收集正常与退变椎间盘组织，一部分用于髓核细胞培养，观察不同退变等级的髓核细胞的生长状态;一部分提取核蛋白应用Western blot方法进一步确认NF-κB信号通路在退变椎间盘组织中的表达。使用激光共聚焦显微镜检测不同退变等级髓核细胞中NF-κB-p65在细胞核中的表达，明确NF-κB信号通路在退变椎间盘组织中的活性的变化。采用实时定量PCR方法检测不同退变等级椎间盘组织中蛋白多糖，Ⅱ型胶原，MMPs及ADAMTS的表达量。　　结果:成功培养人髓核细胞，倒置显微镜下观察，未退变髓核细胞大部分为短梭形、多边形，且细胞体积比较大。胞质丰富，具有折光性，胞核大且呈卵圆形;退变髓核细胞逐渐以梭形为主，胞质减少，折光性降低，生长速度减慢。荧光染色观察各组髓核细胞胞核中p65的变化:未退变髓核细胞显示出p65蛋白细胞质定位（指示未激活的NF-κB），退变髓核细胞p65蛋白主要集中在细胞核中(指示激活状态的NF-κB)，与未退变髓核细胞相比胞质荧光强度较弱。免疫印迹检测各组髓核组织中核蛋白p65蛋白的表达:与未退变髓核组织相比，退变的髓核组织中核蛋白p65的表达随着退变等级的增高逐渐增高。NF-κB调控靶基因RT-PCR结果:退变组基质金属蛋白酶(MMP-1、MMP-3、MMP-13)，聚集蛋白聚糖酶(ADAMTS-4、ADAMTS-5)和IL-6的mRNA表达明显高于未退变组;聚集蛋白聚糖和Ⅱ型胶原较未退变组明显降低。　　结论:在椎间盘发生退变时，NF-κB活性逐渐增强，调控一些基质降解酶，如:基质金属蛋白酶和聚集蛋白聚糖酶的高表达，导致细胞外基质的降解的加剧。　　第二部分:NF-κB信号通路在椎间盘退变中的机制研究　　研究背景:椎间盘退变(intervertebral disc degeneration，IDD)所继发的腰椎间盘突出症、腰椎管狭窄症、盘源性腰痛和腰椎不稳症等疾患，是腰腿痛的主要病因。临床上表现为发病率高、涉及人群广、易复发，是严重影响人类健康的一大类疾患，目前无有效的治疗措施，同时给社会带来巨大的经济负担。　　椎间盘退变是一个多因素协同作用、不断进展的慢性过程。虽然椎间盘退变的原因和病理生理学仍不清楚，

关键词： Anterior-posterior polarity   ASPN   Eye induction   IGF   Xenopus  

摘要： Small leucine-rich repeat proteoglycan (SLRP) family proteins play important roles in a number of biological events. Here, we demonstrate that the SLRP family member Asporin (ASPN) plays a crucial role in the early stages of eye development in Xenopus embryos. During embryogenesis, ASPN is broadly expressed in the neuroectoderm of the embryo. Overexpression of ASPN causes the induction of ectopic eyes. By contrast, blocking ASPN function with a morpholino oligonucleotide (ASPN-MO) inhibits eye formation, indicating that ASPN is an essential factor for eye development. Detailed molecular analyses revealed that ASPN interacts with insulin growth factor receptor (IGFR) and is essential for activating the IGF receptor-mediated intracellular signalling pathway. Moreover, ASPN perturbed the Wnt, BMP and Activin signalling pathways, suggesting that ASPN thereby creates a favourable environment in which the IGF signal can dominate. ASPN is thus a novel secreted molecule essential for eye induction through the coordination of multiple signalling pathways. [ABSTRACT FROM AUTHOR]

关键词： Asporin   蛋白多糖   结直肠癌   癌基因  

摘要： 目的探究Asporin在结直肠癌及癌旁正常组织的表达差异。方法选择32例结直肠癌及其对应癌旁正常黏膜组织的石蜡标本,使用免疫组化方法检测Asporin的表达;12例新鲜切取的结直肠癌组织及配对癌旁正常组织通过Western Blot检测Asporin的表达。比较癌组织及癌旁正常组织Asporin表达的差异。结果免疫组织化学结果显示,相对于癌旁组织,Asporin在结直肠肿瘤组织中异常高表达,32例结直肠癌Asporin高表达率为68.75%(22/32)。蛋白免疫印迹结果显示,12例新鲜肿瘤标本发现有9例Asporin呈高表达(P<0.01)。结论 Asporin可能做为一种癌基因,参与结直肠癌的发生发展。

关键词： Asporin   Single nucleotide polymorphism   Knee osteoarthritis  

摘要： The aim of this study was to investigate associations between single nucleotide polymorphisms rs13301537 in asporin (ASPN) and rs373444 in the bone morphogenetic protein 5 (BMP5) gene with knee osteoarthritis (OA) susceptibility in a Chinese Han population. ASPN rs13301537 and BMP5 rs373444 polymorphisms were genotyped in patients with knee OA and age- and sex-matched OA-free controls from a Chinese Han population. A total of 510 patients with knee OA and 520 controls were enrolled in the study. CT and CC genotypes of rs13301537, and variant C, were associated with a significantly increased risk of knee OA. On stratification analysis, the association between the risk of OA and rs13301537 CT heterozygotes compared with TT homozygotes was stronger in females and those aged > 65 years. In contrast, the CT and CC genotypes of rs373444 in BMP5 were not significantly associated with the risk of knee OA, even after further stratification analysis according to age or sex. Our results showed that ASPN rs13301537 T to C change and variant C genotype may contribute to knee OA risk in a Chinese Han population.

关键词： 腰椎   椎间盘退行性变   基因疗法   综述  

摘要： 下腰痛是一种常见病、多发病，它影响人们的生活质量和精神状态，成为困扰人类的疾病之一［1-3］。很多因素都会导致下腰痛，其中椎间盘退变是最重要的因素之一［4］。目前临床上针对椎间盘退变引起相关疾病的治疗策略是解除椎间盘退变相关的脊髓、神经和血管刺激或压迫，并非针对椎间盘退变的病理过程，因而临床症状可反复发作，而且手术后病变节段的相邻节段椎间盘退变加速［5-6］。随着科学技术的发展，特别是分子生物学技术不断进步，延缓甚至逆转椎间盘退变正成为可能。目前治疗椎间盘退变的分子生物学技术主要包括三大类：细胞治疗、组织工程治疗及基因治疗。本综述主要着重基因治疗。

关键词： 椎间盘退变   Asporin   TGF-β1   L929  

摘要： 目的:构建高表达asporin基因L929细胞株,研究其对子小鼠正常成纤维细胞L929 TGF-β1及Col2α1表达的影响。方法:运用RT-PCR技术从小鼠下颌骨组织中扩增asporin cDNA,连接酶连接至真核表达载体pcDNA3.1(-),所得的重组质粒pcDNA3.1(-)-ASPNⅢ用脂质体法转染小鼠成纤维细胞L929,经过G418筛选,使用RT-PCR以及Western blot技术筛选asporin基因高表达的细胞株。并使用RT-PCR以及Western blot技术检测asporin基因高表达对细胞TGF-β1及Col2α1表达的影响。结果 :成功获得asporin高表达的L929细胞株。RT-PCR以及Western blot技术检测高表达asporin对TGF-β1及Col2α1的表达有明显的抑制作用。结论:asporin基因在L929细胞中高表达能抑制细胞TGF-β1及Col2α1的表达。