关键词： 腰椎间盘退变   基因治疗   非手术治疗   治疗方法   椎间盘再生   医务工作者   分子生物学   介入治疗  

摘要： 腰椎间盘退变及其继发的相关组织异常改变所致下腰痛是临床极为常见的病症，每年要消耗大量的人力、物力和财力。治疗方法有非手术治疗、手术治疗和介入治疗等。手术治疗会破坏脊柱运动单位的生理完整性，而且不能从根本上阻止椎间盘退变的发生和发展。因此如何在保持脊柱力学结构和生理完整性的前提下进行椎间盘退变的治疗，即在早期阻止或逆转椎间盘退变，促进椎间盘再生，是医务工作者目前集中研究的课题之一。随着分子生物学的飞速发展，采用基因治疗阻止或逆转椎间盘退变已成为有应用前景的治疗方法。

关键词： 椎间盘退变   分子机制   基因治疗  

摘要： 椎间盘退变是下腰痛的常见病因。椎间盘退变分子机制的研究发现,白介素-1α和β、肿瘤坏死因子-α、一氧化氮、环氧化酶-2、基质金属蛋白酶等细胞因子和炎症介质在退变椎间盘中有较高活性,转化生长因子-β及Sox9基因等表达降低,可能是椎间盘退变发生发展的重要原因之一。基因治疗为椎间盘退变带来了希望,将骨形态发生蛋白-2和骨诱导蛋白-1基因,转化生长因子-β、胰岛素样生长因子-1和生长分化因子-5等生长因子基因,以及Sox9基因成功导入人或动物椎间盘细胞中发现,这些外源基因可促进Ⅱ型胶原和(或)蛋白多糖不同程度的表达上调;将白介素-1受体拮抗剂和组织基质金属蛋白酶抑制因子-1基因成功导入椎间盘细胞,可从抑制分解代谢方面为基因治疗提供另一条可行的途径。基因治疗也存在一些问题与不足。随着研究的深入与转基因技术方法的提高,基因治疗可望进入临床为椎间盘退变开辟新的治疗途径。

关键词： Animals   Extracellular Matrix Proteins   Humans   Mice   Osteoarthritis   Promoter Regions (Genetics)  

摘要： Asporin ( ASPN) is a small leucine-rich proteoglycan that is involved in pathological processes of osteoarthritis. Previously, we showed that asporin can inhibit transforming growth factor-beta 1 ( TGF-beta 1)-mediated expression of cartilage matrix genes and chondrogenesis in vitro (Kizawa, H., Kou, I., Iida, A., Sudo, A., Miyamoto, Y., Fukuda, A., Mabuchi, A., Kotani, A., Kawakami, A., Yamamoto, S., Uchida, A., Nakamura, K., Notoya, K., Nakamura, Y., and Ikegawa, S. ( 2005) Nat. Genet. 37, 138 -144). However, details about regulation of asporin itself are not yet known. Here, we examined ASPN expression in skeletal tissue and potential regulation of ASPN by TGF-beta. In situ hybridization revealed the presence of ASPN mRNA in the perichondrium/periosteum of long bones, but its absence in articular cartilage and growth plates. Immunohistochemical analysis also showed ASPN protein expression predominantly in the perichondrium/periosteum. TGF-beta 1 induced endogenous ASPN mRNA expression over time in vitro, and this induction was suppressed by the TGF-beta type I receptor kinase inhibitor SB431542. Inhibition of Smad3 significantly reduced TGF-beta 1induced ASPN expression, whereas overexpression of Smad3 augmented the induction. Characterization of the human ASPN promoter region revealed a region from -126 to -82 that is sufficient for full promoter activity;however, TGF-beta 1 failed to increase activity through the ASPN promoter. Our findings indicate that TGF-beta 1 induces ASPN through Smad3 but that this induction is indirect.

关键词： Cartilage Articular   Cell Differentiation   Cell Line   Extracellular Matrix Proteins  

摘要： Osteoarthritis ( OA), the most prevalent form of skeletal disease, represents a leading cause of disability following middle age. OA is characterized by the loss of articular cartilage;however, the details of its etiology and pathogenesis remain unclear. Recently, we demonstrated a genetic association between the cartilage extracellular matrix protein asporin and OA ( Kizawa, H., Kou, I., Iida, A., Sudo, A., Miyamoto, Y., Fukuda, A., Mabuchi, A., Kotani, A., Kawakami, A., Yamamoto, S., Uchida, A., Nakamura, K., Notoya, K., Nakamura, Y., and Ikegawa, S. ( 2005) Nat. Genet. 37, 138-144). Furthermore, we showed that asporin binds to transforming growth factor-beta (TGF-beta), a key cytokine in OA pathogenesis, and inhibits TGF-beta-induced chondrogenesis. To date, functional data for asporin have come primarily from mouse cell culture models of developing cartilage rather than from human articular cartilage cells, in which OA occurs. Here, we describe mechanisms for asporin function and regulation in human articular cartilage. Asporin blocks chondrogenesis and inhibits TGF-beta 1-induced expression of matrix genes and the resulting chondrocyte phenotypes. Small interfering RNA-mediated knockdown of asporin increases the expression of cartilage marker genes and TGF-beta 1;in turn, TGF-beta 1 stimulates asporin expression in articular cartilage cells, suggesting that asporin and TGF-beta 1 form a regulatory feedback loop. Asporin inhibits TGF-beta/Smad signaling upstream of TGF-beta type I receptor activation in vivo by co-localizing with TGF-beta 1 on the cell surface and blocking its interaction with the TGF-beta type II receptor. Our results provide a basis for elucidating the role of asporin in the molecular pathogenesis of OA.

关键词： 椎间盘退变   基因治疗   脊柱退变性疾病   疾病过程   椎间盘突出   神经根病变   disc   脊柱不稳  

摘要： 椎间盘退变(intervertebral disc degeneration,IDD)可以引起一系列脊柱退变性疾病如椎间盘突出、脊柱不稳、神经根病变和脊髓病变等,目前的治疗方法主要是手术治疗,但手术方法只是处理终末期的症状,不能逆转疾病过程本身。因此国内外学者们一直致力于寻找一种能逆转这个疾病过程的方法,近来对椎间盘的生化构成及其内环境的进一步了解以及某些基因被发现对椎间盘内基质合成和分解有着重要的影响,这些为科学家寻求逆转这个疾病过程提供了新方法。[第一段]

关键词： Animals   Calcification Physiologic   Cell Differentiation   Cell Line   Cloning Molecular  

摘要： Periodontal ligament-associated protein-1 (PLAP-1)/asporin is a recently identified novel member of the small leucine-rich repeat proteoglycan family. PLAP-1/asporin is involved in chondrogenesis, and its involvement in the pathogenesis of osteoarthritis has been suggested. We report that PLAP-1/asporin is also expressed specifically and predominantly in the periodontal ligament (PDL) and that it negatively regulates the mineralization of PDL cells. In situ hybridization analysis revealed that PLAP-1/asporin was expressed specifically not only in the PDL of an erupted tooth but also in the dental follicle, which is the progenitor tissue of the PDL during tooth development. Overexpression of PLAP-1/asporin in mouse PDL-derived clone cells interfered with both naturally and bone morphogenetic protein 2 (BMP-2)-induced mineralization of the PDL cells. On the other hand, knockdown of PLAP-1/asporin transcript levels by RNA interference enhanced BMP-2-induced differentiation of PDL cells. Furthermore co-immuno-precipitation assays showed a direct interaction between PLAP1/asporin and BMP- 2 in vitro, and immunohistochemistry staining revealed the co-localization of PLAP-1/asporin and BMP- 2 at the cellular level. These results suggest that PLAP-1/asporin plays a specific role(s) in the periodontal ligament as a negative regulator of cytodifferentiation and mineralization probably by regulating BMP- 2 activity to prevent the periodontal ligament from developing non-physiological mineralization such as ankylosis.

关键词： asporin   association   knee osteoarthritis   age at onset   Han Chinese  

摘要： Recent genetic studies for osteoarthritis (OA) have been focused on ASPN, the gene encoding asporin, where aspartic acid (D)-repeat polymorphisms are associated with OA in several ethnic groups. The purpose of the present study is to further examine the association of the D-repeat polymorphism in a Han Chinese population. The D-repeat polymorphism was genotyped in 354 knee OA patients (257 women and 97 men) who suffered from primary symptomatic knee OA with radiographic confirmation, and the association of the repeat with various clinical parameters was examined. The age at onset (years) in patients with the D14 allele (mean: 51.9, SD: 8.5) was younger than that those without the D14 allele (mean: 54.9, SD: 10.9) (P = 0.023). Survival analysis showed D14 was significantly associated with age at onset of OA (P = 0.004 in the dominant model). The average age at onset of patients with the D13/D13 genotype (mean: 56.1, SD: 11.1) was older than those without the D13/D13 genotype (mean: 53.0, SD: 9.9) (P = 0.013). Our study further highlighted the significance of asporin in OA.

关键词： 人类椎间盘退变   分子生物学机制   治疗前   非类固醇抗炎药   基因   脊柱长度   椎体连接   尸体标本  

摘要： 人类的脊柱由23个椎间盘将椎体连接在一起，从颈椎到腰椎，椎间盘约占脊柱长度的20％～30％，其有着使脊柱具有活动性及缓冲各向应力的作用。人类椎间盘退变从20岁开始，在尸体标本上，在49岁时，大约有97％的椎间盘已经退变或有退变迹象。目前对椎间盘退变所引起的疾病主要的治疗观点是：（1）保守治疗，应用非类固醇抗炎药（NSAIDs）等控制炎症反应；

关键词： Rheumatology  

摘要： Background: Asporin belongs to a family of proteins associated with the cartilage matrix. Objective: To investigate the role of the functional polymorphism consisting of an aspartic acid (D) repeat polymorphism located in the ASPN gene in the susceptibility to and clinical outcome of rheumatoid arthritis. Methods: A total of 803 Spanish Caucasian patients with rheumatoid arthritis and 904 controls of the same ethnic origin and matched for age and sex were included in the study. The asporin D repeat polymorphism was genotyped using polymerase chain reaction with a fluorescent primer. Results: No significant differences were detected in the distribution of the 10 alleles found in our population on comparing patients with rheumatoid arthritis with control groups. Nevertheless, individuals bearing D14 produced rheumatoid factor more often than the rest (85.7% v 72.1%, p = 0.006, odds ratio (OR) = 2.35, 95% confidence interval 1.21 to 4.50), and the mean (SD) onset age was higher in the group of individuals bearing D13 (50.09 (13.94)) compared with the rest (47.21 (14.31)), although the difference did not reach significance (p = 0.06). Conclusion: The results do not support a major role for asporin D repeat polymorphism in the susceptibility to rheumatoid arthritis. Nevertheless, they support the influence of this gene on the outcome of the disease.