• Journal of Acupuncture Research
• /
• v.22 no.2
• /
• pp.71-81
• /
• 2005

Objective : Dear antler (Cervus korean TEMMINCK var. mantchuricus Swinhoe) used for traditional immunosuppressive and immuno-activating action. The effect of deer antler herbal-acupuncture(DAH) solution, prepared by water extract method, on cathepsin activities in bone tissues (cartilage and synovial) cells from mouse rheumatoid arthritis (RA) model was studied. The cysteine endoprotease cathepsin mediates degradation of the MHC class II invariant chain (Ii) in human and mouse antigen-presenting cells. The studies described here examine the functional significance of cathepsin inhibition on autoantigen presentation and organ-specific autoimmune diseases in a murine model for RA. Methods : An animal model for RA in BALB/c mice thymectomized 3 days after birth (3d-Tx) was constructed All 3d-Tx BALB/c mice developed autoimmune lesions in the bone tissue cells, starting at 3 weeks of age, and the disease mediated by CD4+ T cells was chronic and progressive. Significant inhibitory effects of DAH solution on cathepsin S and L were observed in each organ in a dose-dependent manner. Moreover, we confirmed that cathepsin S and L activity in each organ were clearly inhibited by DAB solution. When we examined the inhibitory effects of DAH solution against autoantigen-specific T cell responses in vitro, in regional lymph node cells, but not in spleens, from model mice, a significant inhibitory effect of DAB solution was observed in a dose-dependent manner. DAH solution do not block T cell proliferation to Con A, indicated that the dose of DAB solution 10 to $20\;{\mu}g/m{\ell}$ was sufficient to inactivate the autoantigen-specific T cell responses in vitro. In vivo therapeutic effects of DAB solution were examined in a murine model for RA, autoantigen-specific (C-II-specific) T cell response were significantly inhibited in LNCs from DAH solution-treated mice. Results : Iinhibition of cathepsin S and L in vivo alters autoantigen presentation and development of organ-specific autoimmunity in RA model. Conclusion : These data identify selective inhibition of cysteine protease cathepsin S and L as a potential therapeutic strategy for autoimmune disease process such RA. Thus, DAH solution will served as a potent anti-inflammatory and anti-arthritic agents for treatment of human RA.

• Journal of dental hygiene science
• /
• v.6 no.1
• /
• pp.1-9
• /
• 2006

Cathepsin K (cat K) is the major cysteine protease expressed in osteoclasts and was thought to play a key role in matrix degradation during bone resorption. It was shown that the intracellular maturation of cat K was prevented by the cAMP antagonist, Rp-cAMP, and the protein kinase A (PKA) inhibitors of KT5720 and H89. In contrast, forskolin, a adenylate cyclase agonist, rather induced Cat K processing and maturation in osteoclasts. Furthermore, to determine whether cat K processing and maturation signaling involves protein kinase C (PKC), mouse total bone cells were treated with calphostin C, a specific inhibitor of PKC, however, no effect was observed, indicating that calphostin C did not affect to osteoclast-mediated cat K processing and maturation. Thus, it is indicated that the cAMP-PKA signaling pathway regulates cat K maturation in osteoclasts. Since secreted proenzymes have the potential to reenter the cell via M6P receptor, to prevent this possibility, it was tested cAMP antagonist Rp-cAMP and the PKA inhibitors KT5720 and H89 in the absence or presence of M6P. Inhibition of cat K processing by Rp-cAMP, KT5720, or H89 was observed in a dose-dependent manner. Furthermore, the addition of M6P resulted in enhanced potency of Rp-cAMP, KT5720 and H89. These dose-dependently inhibited in vitro bone resorption with a potency similar to that observed for inhibition of cat K processing.

• Development and Reproduction
• /
• v.17 no.2
• /
• pp.133-140
• /
• 2013

This study was performed to investigate the expression of cathepsin B mRNA and protein in eutopic and ectopic endometrial tissues of patients with endometriosis and in normal endometrial tissues and to clarify the association between the cathepsin B expression and endometriosis. A total of 40 women with histologically confirmed endometriosis were recruited for study group. For controls, 20 women undergoing operative treatment for uterine myoma, cervical intraepithelial neoplasia (CIN) or benign gynecologic conditions other than endometriosis were recruited. Eutopic endometrial tissues of both groups and ectopic endometrial tissue of study group were collected during the operations. We employed real time reverse transcriptase - polymerase chain reaction (RT-PCR) to quantify mRNA levels of cathepsin B in these tissues. Then, we performed western blot analysis to measure the protein levels of cathepsin B. The expressions of cathepsin B mRNA and protein were significantly higher in both eutopic and ectopic endometrial tissues of women with endometriosis than in endometrial tissues of controls. These data suggest that the higher expression of cathepsin B in the endometrial tissues might be associated with the development of endometriosis. In addition, eutopic endometrium itself with higher expression cathepsin B may play a pivotal role in the histogenesis of endometriosis.

• Proceedings of the PSK Conference
• /
• 2003.10b
• /
• pp.180.2-180.2
• /
• 2003

Cathepsin K, a cysteine protease of the papain superfamily, is predominantly expressed in osteoclasts and has been postulated as a target for the treatment of osteoporosis. Crystallographic and structure-activity studies on a series of azepanone-based diamino and acyclic ketone derivative inhibitors of cathepsin K have led to the design and identification. X-ray structure of the cysteine protease cathepsin K (1NL6) co-crystalized with an inhibitor with 2.8${\AA}$ resolution was used to predict the protein-ligand interactions and to estimate the binding affinity from the docking score by FlexX module. (omitted)

• Journal of Microbiology and Biotechnology
• /
• v.18 no.6
• /
• pp.1115-1120
• /
• 2008

A novel cathepsin B inhibitor-producing bacterium was isolated from marine sediments and identified based on its 16S rDNA sequence as Pseudomonas sp. strain PB01 (Accession No. EU126129). The growth and enzyme inhibitor production were investigated under various culture conditions. A mixture of organic nitrogen source was required for the optimal production, whereas both glucose and maltose proved to be the effective carbon sources for cathepsin B inhibitor production. Other optimal culture conditions included temperature range between 25 and $28^{\circ}C$, initial medium pH of 6.6, and shaking speed of 200 rpm. Under these optimal conditions, the maximum inhibitory activity from culture broth was approximately 50% after 30 h of cultivation. Additionally, kinetic study revealed that inhibitor production paralleled with cell growth, which suggested that the inhibitor may be a primary metabolite of that bacterium.

• Journal of Gastric Cancer
• /
• v.2 no.3
• /
• pp.151-156
• /
• 2002

Purpose: The prognosis of gastric cancer depends on the depth of invasion, lymph-node metastasis, invasion to adjacent tissues, and distant metastasis. Recently, it is known that tumor-associated proteases and adhesion molecules have been shown to play a relevant role in the process of progression and metastasis. The purpose of our study was to demonstrate the value of MMP-2 (matrix metalloproteinase), cathepsin D and E-cadherin as prognostic factors. Materials and Methods: In this study, formalin-fixed, paraffin-embedded tissue blocks from 69 patients with gastric cancer were immunohistochemically studied using antibodies to MMP-2, cathepsin D, and E-cadherin, and their expressions were analyzed according to the pathologic stage, lymph-node metastasis, histological differentiation, and patient survival. The medical records of these patients were retrospectively reviewed. Results: Increased expression of MMP-2 significantly correlated with advanced pathologic stage (P=0.026). Patients with lymph-node metastasis also had increased expression of MMP-2. Those patients with increased expression of MMP-2 showed a poorer survival; nevertheless, it was not statistically significant. Increased expression of cathepsin D significantly correlated with advanced pathologic stage (p=0.029). However, no correlation was observed between advanced pathologic stage and either lymph-node status or histological differentiation. Patients with increased expression of cathepsin D had a poorer survival, but that result was not statistically significant. No association was found between reduced expression of E-cadherin and pathologic stage, lymph-node status, or histological differentiation. Also, no correlation was found between the expression of E-cadherin and survival. In addition, when a combination of MMP-2 and cathepsin D expressions was analyzed, if both were negative, the survival seems to be longer, but it was not statistically significant. Conclusions: In patients with gastric cancer, expressions of MMP-2 and cathepsin D correlated with tumor stage; therefore, they may be considered as prognostic factors.

• Journal of Fisheries and Marine Sciences Education
• /
• v.28 no.4
• /
• pp.903-912
• /
• 2016

Hagfish which belongs to the chordate contact cyclostomata, is important phylogenetic relationship between vertebrate and invertebrate. Cathepsins of the cysteine protease family have traditionally been thought to play a major role in intracellular protein degradation and turnover in lysosomes. In this study, Catepsin L was cloned from Inshore hagfish (Eptatretus burgeri), the cDNA encoding ORF of the Eptatretus burgeri Cathepsin L (EbCtL) is 978 bp. The cDNA encoding proEbCtL was expressed in Escherichia coli strain BL21(DE3) using the pGEX-4T-1 expression vector system. The recombinant proEbCtL protein was overexpressed as a approximately 55 kDa fusion protein. The overproduced soluble GST-fusion protein was then applied to glutathione-Sepharose 4B column chromatography; the sample harboring the fusion protein evidenced a high degree of purity when analyzed via SDS-PAGE and Western blot analysis. Its activity was quantied by cleaving the synthetic peptide Z-FR-AMC, Z-LLE-AMC, and Suc-AAF-AMC, and the optimal pH for the protease activity was 8, 9.5, and 9, respectively.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 2001.04a
• /
• pp.89-99
• /
• 2001

■ Cathepsin K is a attractive target for selectively and efficiently modulating the osteoclastic bone resorption. ■ OST-1857 is a lead compound which is specifically targeted to cathepsin K and showed efficacy in TPTX rats. ■ OST-compounds are in process of the preclinical study, joined by Yuhan research center.

• Restorative Dentistry and Endodontics
• /
• v.44 no.2
• /
• pp.17.1-17.10
• /
• 2019

Objectives: Root resorption is an unexpected complication after replantation procedures. Combining anti-osteoclastic medicaments with retrograde root filling materials may avert this resorptive activity. The purpose of this study was to assess effects of a cathepsin K inhibitor with calcium silicate-based cements on osteoclastic activity. Methods: MC3T3-E1 cells were cultured for biocompatibility analyses. RAW 264.7 cells were cultured in the presence of the receptor activator of nuclear factor-kappa B and lipopolysaccharide, followed by treatment with Biodentine (BIOD) or ProRoot MTA with or without medicaments (Odanacatib [ODN], a cathepsin inhibitor and alendronate, a bisphosphonate). After drug treatment, the cell counting kit-8 assay and Alizarin red staining were performed to evaluate biocompatibility in MC3T3-E1 cells. Reverse-transcription polymerase chain reaction, tartrate-resistant acid phosphatase (TRAP) staining and enzyme-linked immunosorbent assays were performed in RAW 264.7 cells to determine the expression levels of inflammatory cytokines, interleukin $(IL)-1{\beta}$, IL-6, tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) and prostaglandin E2 (PGE2). Data were analyzed by one-way analysis of variance and Tukey's post hoc test (p < 0.05). Results: Biocompatibility results showed that there were no significant differences among any of the groups. RAW 264.7 cells treated with BIOD and ODN showed the lowest levels of $TNF-{\alpha}$ and PGE2. Treatments with BIOD + ODN were more potent suppressors of inflammatory cytokine expression (p < 0.05). Conclusion: The cathepsin K inhibitor with calcium silicate-based cement inhibits osteoclastic activity. This may have clinical application in preventing inflammatory root resorption in replanted teeth.

• International Journal of Oral Biology
• /
• v.42 no.3
• /
• pp.137-142
• /
• 2017

To determine the effect of the tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) in odontoclast formation, we administrated a $TNF-{\alpha}$ inhibitor in rats with diabetes rats with periodontitis. The rats included in the study were divided into three groups: control rats without diabetes or periodontitis (the C group), rats with periodontitis and diabetes (the PD group), and rats with periodontitis and diabetes treated by infliximab, the TNF inhibitor (the PD+infliximab group). The PD and PD+infliximab groups received intravenous administrations of streptozotocin (STZ, 50 mg/kg) to induce diabetes. After 7 days of STZ injections, the mandibular first molars were ligatured to induce periodontitis. The PD+infliximab group was intrapenitoneally administrated by infliximab (5 mg/kg). On days 3 and 20 after the ligature administration, odontoclast formation along root surfaces was evaluated by tartrate resistant acid phosphatase (TRAP) staining and cathepsin K immunohistochemistry. On day 3, the number of TRAP- and cathepsin K-positive cells increased more so in the PD group than in the C group. The PD+infliximab group showed a lower number of positive cells than the PD group. There was no difference in all the groups on day 20. On day 3, the cathepsin-K positive multinucleated and mononucleated cells were higher in the PD group than in the C group. The number of cathepsin-K positive multinucleated cells was lower in the PD+infliximab group than in the PD group. The PD group showed more cathepsin K-positive cells in the furcation and distal surfaces than the c group. The Cathepsin K-positive cells of the PD+infliximab group were lower than that of the PD group in furcation. These results suggest that $TNF-{\alpha}$ stimulates odontoclast formation in diabetes with periodontitis.