Search

Mesangial matrix expansion is a prominent feature of the most common form of glomerulonephritis, IgA nephropathy (IgAN). To find molecular markers and improve the understanding of the disease, the gene and protein expression of proteoglycans were investigated in biopsies from IgAN patients and correlated to clinical and morphological data. We collected and microdissected renal biopsies from IgAN patients (n = 19) and from healthy kidney donors (n = 14). Patients were followed for an average time of 4 years and blood pressure was according to target guidelines. Distinct patterns of gene expression were seen in glomerular and tubulo-interstitial cells. Three of the proteoglycans investigated were found to be of special interest and upregulated in glomeruli: perlecan, decorin and biglycan. Perlecan gene expression negatively correlated to albumin excretion and progress of the disease. Abundant decorin protein expression was found in sclerotic glomeruli, but not in unaffected glomeruli from IgAN patients or in controls. Transforming growth factor beta (TGF-β), known to interact with perlecan, decorin and biglycan, were upregulated both on gene and protein level in the glomeruli. This study provides further insight into the molecular mechanisms involved in mesangial matrix expansion in IgAN. We conclude that perlecan is a possible prognostic marker for patients with IgAN. In addition, the up-regulation of biglycan and decorin, as well as TGF-β itself, indicate that regulation of TGF-β, and other profibrotic markers plays a role in IgAN pathology.

Objective: Decorin and oxidized low-density lipoprotein (Ox-LDL) independently induce osteogenic differentiation of vascular smooth muscle cells (VSMCs). We aimed to determine whether decorin glycosaminoglycan (GAG) chain synthesis contributes to Ox-LDL-induced differentiation and calcification of human VSMCs in vitro., Methods and Results: Human VSMCs treated with Ox-LDL to induce oxidative stress showed increased alkaline phosphatase (ALP) activity, accelerated mineralization, and a difference in both decorin GAG chain biosynthesis and CS/DS structure compared with untreated controls. Ox-LDL increased mRNA abundance of both xylosyltransferase (XT)-I, the key enzyme responsible for GAG chain biosynthesis and Msx2, a marker of osteogenic differentiation. Furthermore, downregulation of XT-I expression using small interfering RNA blocked Ox-LDL-induced VSMC mineralization. Adenoviral-mediated overexpression of decorin, but not a mutated unglycanated form, accelerated mineralization of VSMCs, suggesting GAG chain addition on decorin is crucial for the process of differentiation. The decorin-induced VSMC osteogenic differentiation involved activation of the transforming growth factor (TGF)-β pathway, because it was attenuated by blocking of TGF-β receptor signaling and because decorin overexpression potentiated phosphorylation of the downstream signaling molecule smad2., Conclusions: These studies provide direct evidence that oxidative stress-mediated decorin GAG chain synthesis triggers TGF-β signaling and mineralization of VSMCs in vitro.

Decorin is a small leucine-rich proteoglycan (SLRP) that plays a vital role in many important cellular processes in several tissues including the cornea. A normal constituent of the corneal stroma, decorin is also found in the majority of connective tissues and is related structurally to other small proteoglycans. It interacts with various growth factors such as epidermal growth factor (EGF) and transforming growth factor beta (TGFβ) to regulate processes like collagen fibrillogenesis, extracellular matrix (ECM) compilation, and cell-cycle progression. Studies have linked decorin dysregulation to delayed tissue healing in patients with various diseases including cancer. In the cornea, decorin is involved in the regulation of transparency, a key function for normal vision. It has been reported that mutations in the decorin gene are associated with congenital stromal dystrophy, a disease that leads to corneal opacity and visual abnormalities. Decorin also antagonizes TGFβ in the cornea, a central regulatory cytokine in corneal wound healing. Following corneal injury, increased TGFβ levels induce keratocyte transdifferentiation to myofibroblasts and, subsequently, fibrosis (scarring) in the cornea. We recently reported that decorin overexpression in corneal fibroblasts blocks TGFβ-driven myofibroblast transformation and fibrosis development in the cornea in vitro suggesting that decorin gene therapy can be used for the treatment of corneal scarring in vivo.

Accumulation of connective tissue is a typical feature of chronic liver diseases. Decorin, a small leucine-rich proteoglycan, regulates collagen fibrillogenesis during development, and by directly blocking the bioactivity of transforming growth factor-β1 (TGFβ1), it exerts a protective effect against fibrosis. However, no in vivo investigations on the role of decorin in liver have been performed before. In this study we used decorin-null (Dcn-/-) mice to establish the role of decorin in experimental liver fibrosis and repair. Not only the extent of experimentally induced liver fibrosis was more severe in Dcn-/- animals, but also the healing process was significantly delayed vis-à-vis wild-type mice. Collagen I, III, and IV mRNA levels in Dcn-/- livers were higher than those of wild-type livers only in the first 2 months, but no difference was observed after 4 months of fibrosis induction, suggesting that the elevation of these proteins reflects a specific impairment of their degradation. Gelatinase assays confirmed this hypothesis as we found decreased MMP-2 and MMP-9 activity and higher expression of TIMP-1 and PAI-1 mRNA in Dcn-/- livers. In contrast, at the end of the recovery phase increased production rather than impaired degradation was found to be responsible for the excessive connective tissue deposition in livers of Dcn-/- mice. Higher expression of TGFβ1-inducible early responsive gene in decorin-null livers indicated enhanced bioactivity of TGFβ1 known to upregulate TIMP-1 and PAI-1 as well. Moreover, two main axes of TGFβ1-evoked signaling pathways were affected by decorin deficiency, namely the Erk1/2 and Smad3 were activated in Dcn-/- samples, whereas no significant difference in phospho-Smad2 was observed between mice with different genotypes. Collectively, our results indicate that the lack of decorin favors the development of hepatic fibrosis and attenuates its subsequent healing process at least in part by affecting the bioactivity of TGFβ1., Competing Interests: DISCLOSURE/CONFLICT OF INTEREST The authors declare no conflict of interest.

The purposes of this study were to quantify the secondary proliferation of primordial germ cells (PGCs) in both sexes of rainbow trout, determine if a sex difference in the timing of PGC proliferation and eventual pre-meiotic number exists, and use microarray data collected during this period to identify genes that are associated with PGC mitosis. The experiments used vasa-green fluorescent protein (vasa-GFP) transgenic rainbow trout of known genetic sex that allowed for the identification and collection of PGCs in vivo. An increase was observed in the number of PGCs counted in the gonads of both female and male embryonic vasa-GFP rainbow trout, from 300 to 700° days (water temperature in °C × days post-fertilization). For both sexes, a statistically significant (P < 0.05) increase in the PGC number was first noted at either 350 or 400° days of development. By 700° days, a 20-50-fold increase in germ cell number was apparent. No sex-specific differences in the timing of PGC proliferation or number were notable in any of the families until 700° days. In conjunction, a custom microarray based on cDNA libraries from embryonic rainbow trout gonads was used to identify genes involved in PGC mitosis. Five genes were discovered: guanine nucleotide binding protein, integral membrane protein 2B, transmembrane protein 47, C-src tyrosine-protein kinase, and the decorin precursor protein. All the genes identified have not been previously associated with germ cell mitosis, but are known to be involved with the cell plasma membrane and/or cell signaling pathways., (Copyright © 2011 Wiley-Liss, Inc.)

Background: Juvenile Hyaline Fibromatosis (JHF) is a rare autosomal recessive disorder, histologically characterized by the production and deposition of an unidentified hyaline material in the skin and other organs. Extracellular matrix molecules are implicated in the development of skin lesion which is debilitating and recurrent and, so far, no treatment is satisfactory., Objective: To investigate the expression of matrix metalloproteinases (MMPs), their tissue inhibitors (TIMPs) and proteoglycans in lesional as compared to site-matched lesion-free skin tissue specimens of a JHF patient, aiming to elucidate the aetiopathological mechanisms involved in the development of JHF skin lesions., Methods: Gelatinase activity of MMP-2 and MMP-9 was investigated by gelatine zymography. Protein levels of MMP-2, MMP-9, TIMP-1 and TIMP-2 in skin tissue extracts were measured by ELISA. Gene expression of MMPs, TIMPs and proteoglycans was examined by quantitative RT-PCR., Results: JHF lesions exhibited significantly higher activity as well as elevated protein and gene expression of MMP-2 and MMP-9, as compared to lesion-free skin tissue specimens. Decorin was downregulated and aggrecan was upregulated in lesional skin, as compared to normal skin., Conclusion: The results presented in this study indicate that MMPs and proteoglycans may be involved in the pathogenesis of JHF and therefore these molecules may offer alternative targets for pharmacological intervention to achieve more radical and effective treatment., (Copyright © 2010 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.)

Previous work has shown that versican, decorin and a catabolic fragment of decorin, termed decorunt, are the most abundant proteoglycans in human skin. Further analysis of versican indicates that four major core protein species are present in human skin at all ages examined from fetal to adult. Two of these are identified as the V0 and V1 isoforms, with the latter predominating. The other two species are catabolic fragments of V0 and V1, which have the amino acid sequence DPEAAE as their carboxyl terminus. Although the core proteins of human skin versican show no major age-related differences, the glycosaminoglycans (GAGs) of adult skin versican are smaller in size and show differences in their sulfation pattern relative to those in fetal skin versican. In contrast to human skin versican, human skin decorin shows minimal age-related differences in its sulfation pattern, although, like versican, the GAGs of adult skin decorin are smaller than those of fetal skin decorin. Analysis of the catabolic fragments of decorin from adult skin reveals the presence of other fragments in addition to decorunt, although the core proteins of these additional decorin catabolic fragments have not been identified. Thus, versican and decorin of human skin show age-related differences, versican primarily in the size and the sulfation pattern of its GAGs and decorin in the size of its GAGs. The catabolic fragments of versican are detected at all ages examined, but appear to be in lower abundance in adult skin compared with fetal skin. In contrast, the catabolic fragments of decorin are present in adult skin, but are virtually absent from fetal skin. Taken together, these data suggest that there are age-related differences in the catabolism of proteoglycans in human skin. These age-related differences in proteoglycan patterns and catabolism may play a role in the age-related changes in the physical properties and injury response of human skin.

Imprinted genes play vital roles in the placental development and fetal growth in eutherian mammals. DCN (decorin), PON2 (paraoxonase 2) and PEG3 (paternally expressed 3) genes have been identified as imprinted genes in the mouse. Here, we detected the imprinting status of three genes in the porcine placenta on DG90 (day 90 of gestation) and the expression differences in Yorkshire and Meishan placenta on DG26, DG55 and DG90. The results indicated that the DCN and PON2 genes were not imprinted genes, while the PEG3 gene showed paternal monoallelic expression in porcine placenta. The expression of the DCN gene increased from DG26 to DG90 in both Yorkshire and Meishan pig placenta. However, this gene expression was greater in Yorkshire than Meishan pig on DG55. The expression of the PON2 gene was greater in Meishan pig than that in Yorkshire on DG26 and DG90. The PEG3 gene expression was not affected by day of pregnancy or breed. Data from the present study contribute to function genomic of porcine placental development., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Prion replication in the periphery precedes neuroinvasion in many experimental rodent scrapie models, and in natural sheep scrapie and chronic wasting disease (CWD) in cervids. Prions propagate in the germinal centers of secondary lymphoid organs and are strongly associated with follicular dendritic cells (FDC) and possibly circulating dendritic cells and macrophages. Given the importance of lymphoid organs in prion disease transmission and pathogenesis, gene expression studies may reveal host factors or biological pathways related to prion replication and accumulation. A procedure was developed to enrich for FDC, dendritic cells, and macrophages prior to the investigation of transcriptional alterations in murine splenic cells during prion pathogenesis. In total, 1753 transcripts exhibited fold changes greater than three (false discovery rates less than 2%) in this population isolated from spleens of prion-infected versus uninfected mice. The gene for the small leucine-rich proteoglycan decorin (DCN) was one of the genes most overexpressed in infected mice, and the splenic protein levels mirrored this in mice infected with scrapie as well as bovine spongiform encephalopathy (BSE) and variant Creutzfeldt-Jakob disease (vCJD). A number of groups of functionally related genes were also significantly decreased in infected spleens. These included genes related to iron metabolism and homeostasis, pathways that have also been implicated in prion pathogenesis in the brain. These gene expression alterations provide insights into the molecular mechanisms underlying prion disease pathogenesis and may serve as a pool of potential surrogate markers for the early detection and diagnosis of some prion diseases.

Purpose: Decorin, p57(KIP2), and TGF-beta 1 have been investigated as prognostic factors because they appear to be associated with tumorigenesis; however, the effect of decorin and p57(KIP2) in lung cancer remains poorly understood. The purpose of this study was to examine the expression of decorin, p57(KIP2), and TGF-beta 1 in 64 lung cancer specimens and 36 normal lung specimens, and to analyze the relationships with respect to clinicopathological features and patient survival in lung cancer., Methods: The expression levels of decorin, p57(KIP2), and TGF-beta 1 were examined by in situ hybridization and immunohistochemistry., Results: Normal tissues exhibited a higher expression level of decorin than tumor tissues (p<0.05) and tumor tissues exhibited a higher expression level of TGF-beta 1 than normal tissues (p<0.05). The expression levels of p57(KIP2) and TGF-beta 1 were significantly associated with histological types of lung cancer (p<0.05), and the expression levels of decorin and p57(KIP2) were significantly associated with lymphatic invasion (p<0.05). Moreover, increased expression of decorin and p57(KIP2) correlated with increased survival (decorin, p=0.018; p57(KIP2), p=0.012)., Conclusion: Decreased expression levels of decorin and p57(KIP2) were associated with poor postsurgical survival time and lymphatic metastasis in lung cancer patients; moreover, low expression was an adverse prognostic factor.

Decorin, small leucine-rich proteoglycan, has been shown to modulate angiogenesis in nonocular tissues. This study tested a hypothesis that tissue-selective targeted decorin gene therapy delivered to the rabbit stroma with adeno-associated virus serotype 5 (AAV5) impedes corneal neovascularization (CNV) in vivo without significant side effects. An established rabbit CNV model was used. Targeted decorin gene therapy in the rabbit stroma was delivered with a single topical AAV5 titer (100 µl; 5×10(12) vg/ml) application onto the stroma for two minutes after removing corneal epithelium. The levels of CNV were examined with stereomicroscopy, H&E staining, lectin, collagen type IV, CD31 immunocytochemistry and CD31 immunoblotting. Real-time PCR quantified mRNA expression of pro- and anti-angiogenic genes. Corneal health in live animals was monitored with clinical, slit-lamp and optical coherence tomography biomicroscopic examinations. Selective decorin delivery into stroma showed significant 52% (p<0.05), 66% (p<0.001), and 63% (p<0.01) reduction at early (day 5), mid (day 10), and late (day 14) stages of CNV in decorin-delivered rabbit corneas compared to control (no decorin delivered) corneas in morphometric analysis. The H&E staining, lectin, collagen type IV, CD31 immunostaining (57-65, p<0.5), and CD31 immunoblotting (62-67%, p<0.05) supported morphometric findings. Quantitative PCR studies demonstrated decorin gene therapy down-regulated expression of VEGF, MCP1 and angiopoietin (pro-angiogenic) and up-regulated PEDF (anti-angiogenic) genes. The clinical, biomicroscopy and transmission electron microscopy studies revealed that AAV5-mediated decorin gene therapy is safe for the cornea. Tissue-targeted AAV5-mediated decorin gene therapy decreases CNV with no major side effects, and could potentially be used for treating patients.

Permanent scars form upon healing of tissue injuries such as those caused by ischemia (myocardial infarction, stroke), trauma, surgery, and inflammation. Current options in reducing scar formation are limited to local intervention. We have designed a systemically administered, target-seeking biotherapeutic for scar prevention. It consists of a vascular targeting peptide that specifically recognizes angiogenic blood vessels and extravasates into sites of injury, fused with a therapeutic molecule, decorin. Decorin prevents tissue fibrosis and promotes tissue regeneration by inhibiting TGF-β activity and by other regulatory activities. The decorin-targeting peptide fusion protein had substantially increased neutralizing activity against TGF-β1 in vitro compared with untargeted decorin. In vivo, the fusion protein selectively accumulated in wounds, and promoted wound healing and suppressed scar formation at doses where nontargeted decorin was inactive. These results show that selective targeting yields a tissue-healing and scar-reducing compound with enhanced specificity and potency. This approach may help make reducing scar formation by systemic drug delivery a feasible option for surgery and for the treatment of pathological processes in which scar formation is a problem.

Background: Small leucine-rich repeat proteoglycans (decorin, biglycan, and lumican), collagen, and lymphangiogenesis are involved in tissue remodelling of various organs with inflammatory diseases., Objective: We determined the expression level and the distribution pattern of small leucine-rich repeat proteoglycans (decorin, biglycan, and lumican), collagen and lymphatic vessels in healthy, mild, and severe persistent allergic nasal mucosa., Methods: The distribution pattern of collagen, proteoglycans, and lymphatic vessels in healthy, mild, and severe persistent allergic nasal mucosa was evaluated by the van Gieson staining, immunohistochemistry, RT-PCR, and Western blotting. Quantitative analyses of collagen deposition were calculated as the median of the total percentage area in the tissue specimen. For the evaluation of proteoglycans, the percentage area stained and median optical density were measured for each image. Lymphatic vessels were identified by D2-40 antibody and calculated using the lymphatic vessel density and endothelial length density in tissue specimens. The expression of MMP 2 and 9, TIMP1 and 2 was evaluated with RT-PCR and Western blotting., Results: In mild and severe persistent allergic nasal mucosa, compared with healthy nasal mucosa, collagen showed more intense staining in the superficial and submucosal layer. In healthy and allergic nasal mucosa, decorin was lightly stained without significant differences in the percentage area and optical density of staining. However, lumican and biglycan showed strong immunoreactivity in mild and severe persistent allergic nasal mucosa, which was verified by Western blotting. The number and endothelial length density of lymphatic vessels were increased in mild and severe persistent allergic nasal mucosa compared with healthy nasal mucosa. The expression of MMP 9 was increased in severe persistent allergic rhinitis., Conclusion and Clinical Relevance: These results suggest that the altered distribution pattern of collagen, proteoglycans, and lymphatic vessels could potentially modulate the remodelling of nasal mucosa in mild and severe persistent allergic nasal mucosa., (© 2010 Blackwell Publishing Ltd.)

Decorin (DCN) is a member of small leucine-rich proteoglycan family that neutralizes the bioactivity of transforming growth factor-beta1 (TGF-β1). It has been proven to be a promising anti-fibrotic agent to treat glomerulonephritis. But the underlining mechanism for regulating and degrading intracellular DCN is still not fully understood. In this study, we investigated the roles of ubiquitination in the regulation of cytoplasmic DCN metabolism in rat mesangial cells (MC) by immunoprecipitation and Western blot. The results showed that a proportion of cytoplasmic DCN was ubiquitinated in normal MC and was enhanced in N-glycosylation inhibitor (tunicamycin)-treated MC. After being treated with the proteasome inhibitor MG132, ubiquitinated DCN accumulated and displayed a prolonged half-life, accompanied by decreased TGF-β1 expression and reduced collagen IV mRNA level in MC. This study demonstrated that the stability and function of cytoplasmic DCN can be regulated by ubiquitin-proteasome system (UPS) in MC, which implies that regulating the ubiquitination and degradation of DCN might be a novel approach for modulating MC bioactivity., (© 2010 Wiley-Liss, Inc.)

Purpose: Congenital stromal corneal dystrophy (CSCD) is characterized by stromal opacities that morphologically are seen as interlamellar layers of amorphous substance with small filaments, the nature of which has hitherto been unknown. CSCD is associated with truncating mutations in the decorin gene (DCN). To understand the molecular basis for the corneal opacities we analyzed the expression of decorin in this disease, both at the morphologic and the molecular level., Methods: Corneal specimens were examined after contrast enhancement with cuprolinic blue and by immunoelectron microscopy. Decorin protein from corneal tissue and keratocyte culture was studied by immunoblot analysis before and after O- and N-deglycosylation. The relative level of DCN mRNA expression was examined using Q-RT-PCR, and cDNA was sequenced. Recombinant wild-type and truncated decorin transiently expressed in HEK293 cells were analyzed by gel filtration and immunoblotting., Results: The areas of interlamellar filaments were stained by cuprolinic blue. Immunoelectron microscopy using decorin antibodies revealed intense labeling of these areas. Both wild-type and truncated decorin protein was expressed in corneal tissue and keratocytes of affected persons. When decorin expressed in HEK293 cells was examined by gel filtration, the truncated decorin eluted as high molecular weight aggregates., Conclusions: Accumulation of decorin was found in the interlamellar areas of amorphous substance. The truncated decorin is present in CSCD corneas, and there is evidence it may aggregate in vitro. Thus, decorin accumulation appears to contribute to the stromal opacities that are characteristic of CSCD.

Extracellular matrix (ECM) proteins can modify immune reactions, e.g. by sequestering or displaying growth factors and by interacting with immune and glial cells. Here we quantified by quantitative polymerase chain reaction (qPCR) expression of 50 ECM components and 34 ECM degrading enzymes in multiple sclerosis (MS) active and inactive white matter lesions. COL1A1, COL3A1, COL5A1 and COL5A2 chains were induced strongly in active lesions and even more in inactive lesions. These chains interact to form collagen types I, III and V, which are fibrillar collagens. Biglycan and decorin, which can decorate fibrillar collagens, were also induced strongly. The fibrillar collagens, biglycan and decorin were largely found between the endothelium and astrocytic glia limitans in the perivascular space where they formed a meshwork which was closely associated with infiltrating immune cells. In active lesions collagen V was also seen in the heavily infiltrated parenchyma. Fibrillar collagens I and III inhibited in vitro human monocyte production of CCL2 (MCP-1), an inflammatory chemokine involved in recruitment of immune cells. Together, ECM changes in lesions with different activities were quantified and proteins forming a perivascular fibrosis were identified. Induced fibrillar collagens may contribute to limiting enlargement of MS lesions by inhibiting the production of CCL2 by monocytes.

Objectives: Alteration in the composition of extracellular matrix has been suggested as the major factor for the development of tendinopathy and calcified tendinopathy, which has poorer clinical manifestation. This study investigated the changes of major proteoglycans and collagens in a calcified tendinopathy model and correlated the expression with the acquisition of chondrocyte phenotype, ectopic ossification and loss of matrix organization in the same model., Methods: Thirty-six rats were used. Collagenase or saline was injected into the patellar tendons of each rat. At Weeks 2, 4 and 12, samples were used for immunohistochemistry of major proteoglycans and collagens and mRNA quantification., Results: An increase in collagen type III and I expression was observed after injury at Week 2. Although their levels diminished with time, the ratio of collagen type III to collagen type I remained higher than that in healthy tendon at Week 12. The expression of biglycan, fibromodulin and aggrecan increased with time, whereas the expression of decorin was sustained from Week 2 to Week 12. The expression of major proteoglycans and collagens was observed in the tendon cells and matrix at Week 2 and became localized at the chondrocyte-like cells around the calcific deposits at Week 12., Conclusion: Sustained expression of proteoglycans and a high collagen type III/collagen type I ratio might account for poor matrix organization in calcified tendinopathy. The localization of major proteoglycans around chondro-osseous region might indicate interference of collagen assembly, which favours ectopic chondrogenesis, ossification and predisposition to tendon rupture.

Background/aims: Transforming growth factor beta(1) (TGF-beta(1)) is a key mediator in diabeticnephropathy (DN). Decorin, a natural inhibitor of TGF-beta(1), may have healing properties. We investigated whether overexpression of decorin in the kidneys of streptozocin (STZ)-induced diabetic rats improved pathogenic and clinical changes of DN., Methods: Forty-eight Sprague-Dawley rats were evenly divided into 4 groups: STZ-induced diabetic rats (diabetic-control), decorin adenovirus vector (Ad)-treated STZ rats (Ad-decorin), and Ad-lacZ-treated STZ rats (Ad-lacZ), and vehicle control (PBS-control). At 10, 12, and 16 weeks after STZ treatment, we measured urinary albumin excretion (UAE), and immunolabeled type IV collagen in histological samples of rat kidney. We also measured kidney decorin and TGF-beta(1) levels by reverse transcription polymerase chain reaction and Western blot. Phosphorylated Smad2,3 (p-Smad2,3) was also measured by Western blot., Results: Decorin gene transfection mediated by a recombinant adenovirus has antirenal fibrosis and anti-albuminuria effects in STZ-induced diabetic rats. TGF-beta(1) mRNA and protein expression in diabetic kidney were reduced 2 weeks after Ad-decorin injection., Conclusion: The renal protective effect of decorin in diabetic rats is at least partly due to the downregulation of the TGF-beta(1)/Smad signaling pathway. Ad-decorin shows potential as a therapeutic for human DN., (Copyright 2010 S. Karger AG, Basel.)

To screen differentially expressed genes involved in osteogenic differentiation of human bone marrow stromal cells (BMSCs) at defined stages, subtractive cDNA library was established by means of suppression subtractive hybridization. The BMSCs cultured for 12 and 21 d were used as driver and tester, respectively. A subtract library was successfully constructed and five positive clones were selected from the library. Sequencing analysis and homology comparison showed that the five clones differentially expressed in BMSCs cultured for 21 d were at least 90% homologous with the known genes in human GenBank. It was interestingly found that the osteogenic BMSCs cultured for 21 d differentially expressed decorin and Bax inhibitor 1. RT-PCR was performed to confirm the differentially expressed genes. The results showed that the expression of Bax inhibitor 1 was significantly higher in the cells of 21-day than that of 12-day, while the expression of decorin was only detected in the cells of 21-day.

Objective: To observe the relation between content of type III collagen and expression of decorin in paraurethral connective tissue in patients with stress urinary incontinence (SUI)., Methods: Transvaginal biopsies were obtained from the paraurethral connective tissue in 43 SUI and non-SUI control women. The concentration of type III collagen was determined by immunohistochemical technique, electrophoretic separation and quantification. RT-PCR was used to verify the mRNA level of decorin., Results: The mean concentration of type III collagen was significantly reduced in patients with SUI (23 +/- 4), compared to the control group (34 +/- 6, P < 0.05). The expression of decorin was significantly higher in patients with SUI (1.17 +/- 0.18), compared to the control group (0.76 +/- 0.16, P < 0.05), and it was highly and negatively related to the content of type III collagen (r = -0.720, P < 0.05)., Conclusions: Our study results suggest that collagen metabolism in SUI is altered by decorin. This alteration results in a less flexible connective tissue, which is probably more likely to break. Decorin might be involved in the pathogenesis of SUI.