• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.4
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• pp.532-539
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• 2015

Inhibition of osteoclast differentiation is the most important target for prevention of inflammatory bone resorption and bone diseases. Here, we investigated the effect of spinach ethanol extract on osteoclast differentiation in RAW264.7 cells. Spinach was extracted with ethanol at a concentration ranging from 0 to 100% (0, 25, 50, 75, and 100% ethanol). Inhibitory effects of receptor activator of NF-${\kappa}B$ ligan (RANKL)-induced osteoclast differentiation were evaluated using tartrate-resistant acid phosphatase (TRAP) stain assay. The most effective eanol concentration for osteoclast differentiation was 100%. Spinach extract (100% ethanol) suppressed RANKL-induced osteoclast differentiation and TRAP activity. Spinach extract (100% ethanol) also suppressed expression of osteoclast differentiation-related marker genes (NFATc1, c-FOS, cathepsin K, and TRAP) and down-regulated RANKL-induced NF-${\kappa}B$ and ERK phosphorylation during osteoclast differentiation. Taken together, our results suggest that spinach extract is effective against reducing osteoclast differentiation through the NF-${\kappa}B$-mediated pathway.

• Parasites, Hosts and Diseases
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• v.56 no.5
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• pp.409-418
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• 2018

Acanthamoeba spp. are free-living protozoa that are opportunistic pathogens for humans. Cysteine proteases of Acanthamoeba have been partially characterized, but their biochemical and functional properties are not clearly understood yet. In this study, we isolated a gene encoding cysteine protease of A. castellanii (AcCP) and its biochemical and functional properties were analyzed. Sequence analysis of AcCP suggests that this enzyme is a typical cathepsin L family cysteine protease, which shares similar structural characteristics with other cathepsin L-like enzymes. The recombinant AcCP showed enzymatic activity in acidic conditions with an optimum at pH 4.0. The recombinant enzyme effectively hydrolyzed human proteins including hemoglobin, albumin, immunoglobuins A and G, and fibronectin at acidic pH. AcCP mainly localized in lysosomal compartment and its expression was observed in both trophozoites and cysts. AcCP was also identified in cultured medium of A. castellanii. Considering to lysosomal localization, secretion or release by trophozoites and continuous expression in trophozoites and cysts, the enzyme could be a multifunctional enzyme that plays important biological functions for nutrition, development and pathogenicity of A. castellanii. These results also imply that AcCP can be a promising target for development of chemotherapeutic drug for Acanthamoeba infections.

• Journal of Society of Preventive Korean Medicine
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• v.3 no.1
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• pp.1-33
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• 1999

In order to compare and study the effects of Kamipalmijiwhang tang(加味八味地黃湯) & Kamigangwhalsokdantang(加味羌活續斷湯) on inhibition of osteoporosis induced by ovariectomy, the author administered Kamipalmijiwhangtang & Kamigangwhalsokdantang to the sample groups per orally before and after ovariectomy. The author observed the activities of cathepsin L, K and the contents of calcium, calcitonin, osteocalcin and phosphorus in the serum for the biochemical index, the contents of IL-6 in the serum and bone marrow for immunological index, the state of bone loss in the metaphysis of tibia by histological study, the expression of ELAM-1 &collagen IV by immunohistochemical analysis, the expression of fos by immunocytochemical analysis. The results were as follows: 1. The activities of cathepsin L, K in the serum had a significant inhibitive effects in all sample groups compared with the control group. The inhibitive effects of Sample A-1 & B-1 was better than those of A-2 & B-2 and the inhibitive effects of sample B-1 & B-2 was better than those of A-1 & A-2. 2. The content changes of IL-6 in the serum and bone marrow had a significant inhibitive effects in all sample groups compared with the control group. The inhibitive effects of sample A-1 & B-1 was better than those of A-2 & B-2 and the inhibitive effects of sample B-1 & B-2 was better than those of A-1 & A-2. 3. In the histological study of tibial metaphysis all the sample groups inhibited bone loss significantly compared with the control group and expressed trabecula shape of bone similar to the normal group. 4. The expression of ELAM-1 & collagen IV decreased a little bit in sample A-2. It also decreased to a similar degree in sample B-2 like normal group and disturbed itself. 5. The expression of fos was inhibited in all sample groups compared with control group. The inhibitive expression of sample B-1 & B-2 was more remarkable than those of A-1. & A-2. 6. The serum calcium had a significant increasing effects in an sample groups compared with control group. Each sample groups didn't have any significant difference. 7. The serum calcitonin had a significant increasing effects in sample A-1, A-2 and B-1 groups compared with control group. Each sample groups didn't have any significant difference. 8. The serum osteocalcin and phosphorus didn't have any significant difference in all sample groups compared with control group.

• The Plant Pathology Journal
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• v.36 no.3
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• pp.280-288
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• 2020

RNA interference (RNAi) has attracted attention as a promising approach to control plant viruses in their insect vectors. In the present study, to suppress replication of the rice stripe virus (RSV) in its vector, Laodelphax striatellus, using RNAi, dsRNAs against L. striatellus genes that are strongly upregulated upon RSV infection were delivered through a rice leaf-mediated method. RNAi-based silencing of peroxiredoxin, cathepsin B, and cytochrome P450 resulted in significant down regulation of the NS3 gene of RSV, achieving a transcriptional reduction greater than 73.6% at a concentration of 100 ng/μl and, possibly compromising viral replication. L. striatellus genes might play crucial roles in the transmission of RSV; transcriptional silencing of these genes could suppress viral replication in L. striatellus. These results suggest effective RNAi-based approaches for controlling RSV and provide insight into RSV-L. striatellus interactions.

• BMB Reports
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• v.31 no.4
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• pp.364-369
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• 1998

Insect plasma protein is abundant in the hemolymph of holometabolous insect larvae and is used as a source of amino acids and energy for construction of adult structures during metamorphosis. In order to understand the mechanism of decomposition of larval plasma proteins by hemocyte protease, we tried to purify a cysteine protease from the hemocyte lysate by using Carbobenzoxy-L-Phenylalanyl-L-Arginine-4-Methyl-Coumaryl-7-Amide (Z-Phe-Arg-MCA) as substrate and to identify plasma proteins that are selectively susceptible to the purified protease. Here, we describe the purification and characterization of a cysteine protease that specifically hydrolyzes the plasma protein of the coleopteran insect, Tenebrio molitor, larvae. The molecular mass of this enzyme was 25 kDa, as determined by SDS-PAGE under reducing conditions. The amino acids sequence of its $NH_{2}-terminus$ was determined to be Leu-Pro-Gly-Gln-Ile-Asp-Trp-Arg-Asp-Lys-Gly. This sequence contained Pro, Asp, and Arg residues, conserved in many papain superfamily enzymes. The specific cysteine protease inhibitors, such as E-64 and leupetin, inhibited its hydrolytic activity. One plasma protein with a molecular mass of 48 kDa was selectively hydrolyzed within 3 h when the purified enzyme and plasma proteins were incubated in vitro. However, the 48 kDa protein was not hydrolyzed by the purified 25 kDa protease in the presence of E-64. Western blotting analysis at various developmental stages showed that the purified enzyme was detected at larvae, pupae, and adult stages, but not the embryo stage.

• BMB Reports
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• v.41 no.9
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• pp.670-677
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• 2008

A cDNA microarray composed of 2,028 different ESTs from two shrimp species, Penaeus monodon and Masupenaeus japonicus, was employed to identify yellow head virus (YHV)-responsive genes in hemocytes of P. monodon. A total of 105 differentially expressed genes were identified and grouped into five different clusters according to their expression patterns. One of these clusters, which comprised five genes including cathepsin L-like cysteine peptidase, hypothetical proteins and unknown genes, was of particular interest because the transcripts increased rapidly ($\leq$ 0.25 hours) and reached high expression levels in response to YHV injection. Microarray data were validated by realtime RT-PCR analyses of selected differentially expressed transcripts. In addition, comparative analysis of the hemocyte transcription levels of three of these genes between surviving and non-surviving shrimp revealed significantly higher expression levels in surviving shrimp.

• Preventive Nutrition and Food Science
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• v.10 no.2
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• pp.191-197
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• 2005

There exist two interesting phenomena in making seafood products from surimi. When salted surimi is kept at a constant low temperature $(4\~40^{\circ}C)$, its rheological properties change from sol to gel, which is called 'setting'. Seafood processors can exploit changes that occur during setting in preparation of surimibased products, because heating at high temperatures, after the pre-heating during the setting process, enhances the gel-strength of salted surimi. Contrarily, when salted surimi or low-temperature set gel is heated at moderate temperatures $(50\~70^{\circ}C)$, a deterioration of gel is observed. The phenomenon is termed 'modori'. In the modori temperature range, heat-stable cysteine proteinases such as cathepsin B, H, Land L-Iike hydrolyze the myosins responsible for gel-formation, resulting in gel weakening modori. This article reviews molecular events occurring during gel setting that improve the quality of surimi-based products, and inhibition of modori by applying proteinase inhibitors. Application of recombinant protein technology to surimi-based products is introduced and its prospects for practical use are discussed.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.10a
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• pp.96-96
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• 2018

Although the roots of Heracleum moellendorffii (HM-R) have been long treated for inflammatory human diseases, scientific evidence for the anti-inflammatory activity of HM-R is not sufficient. In this study, we investigated anti-inflammatory activity and mechanism of action of HM-R in LPS-stimulated RAW264.7 cells. HM-R blocked LPS-induced NO and PGE2 production, but not HM-L. HM-R inhibited LPS-induced overexpression of iNOS, COX-2, $IL-1{\beta}$ and IL-6 in RAW264.7 cells. HM-R inhibited LPS-induced $NF-{\kappa}B$ signaling activation through blocking $I{\kappa}B-{\alpha}$ degradation and p65 nuclear accumulation. In addition, HM-R inhibited MAPK signaling activation by attenuating the phosphorylation of ERK1/2, p38 and JNK. Furthermore, HM-R inhibited attenuated LPS-mediated overexpression of the osteoclast-specific factors such as NFATc1, cathepsin K, MCP-1 and TRAP. These results indicate that HM-R may exert anti-inflammatory activity by inhibiting $NF-{\kappa}B$ and MAPK signaling activation. From these findings, HM-R has potential to be a candidate for the development of chemopreventive or therapeutic agents for the inflammation and inflammatory diseases.

• BMB Reports
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• v.57 no.6
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• pp.293-298
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• 2024

Microtubule acetylation has been shown to regulate actin filament dynamics by modulating signaling pathways that control actin organization, although the precise mechanisms remain unknown. In this study, we found that the downregulation of microtubule acetylation via the disruption ATAT1 (which encodes α-tubulin N-acetyltransferase 1) inhibited the expression of RhoA, a small GTPase involved in regulating the organization of actin filaments and the formation of stress fibers. Analysis of RHOA promoter and chromatin immunoprecipitation assays revealed that C/EBPβ is a major regulator of RHOA expression. Interestingly, the majority of C/EBPβ in ATAT1 knockout (KO) cells was found in the nucleus as a 27-kDa fragment (referred to as C/EBPβ^p27) lacking the N-terminus of C/EBPβ. Overexpression of a gene encoding a C/EBPβ^p27-mimicking protein via an N-terminal deletion in C/EBPβ led to competitive binding with wild-type C/EBPβ at the C/EBPβ binding site in the RHOA promoter, resulting in a significant decrease of RHOA expression. We also found that cathepsin L (CTSL), which is overexpressed in ATAT1 KO cells, is responsible for C/EBPβ^p27 formation in the nucleus. Treatment with a CTSL inhibitor led to the restoration of RHOA expression by downregulation of C/EBPβ^p27 and the invasive ability of ATAT1 KO MDA-MB-231 breast cancer cells. Collectively, our findings suggest that the downregulation of microtubule acetylation associated with ATAT1 deficiency suppresses RHOA expression by forming C/EBPβ^p27 in the nucleus through CTSL. We propose that CTSL and C/EBPβ^p27 may represent a novel therapeutic target for breast cancer treatment.

• Journal of Life Science
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• v.24 no.7
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• pp.777-783
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• 2014

Cathepsin D (CtsD), an aspartyl peptidase, is involved in apoptosis, resulting in the release of cytochrome C from mitochondria in cells. Here, we investigated microRNA regulation of CtsD expression in 3T3-L1 cells. First, we observed the expression of CtsD in cells in response to doxorubicin (Dox). As expected, the level of CtsD mRNA increased in 3T3-L1 cells exposed to Dox in a dose-dependent manner. The cellular viability of ectopically expressed CtsD cells was decreased. Next, we used the miRanda program to search for particular microRNA targeting CtsD. MiR-145 was selected as a putative controller of CtsD because it had a high mirSVR score. In a reporter assay, the luciferase activity of cells containing the CtsD 3'-UTR region decreased in cells transfected with a miR-145 mimic compared to that of a control. The level of CtsD expression was down-regulated in preadipocytes ectopically expressing miR-145 and up-regulated by an miR-145 inhibitor. Cells also suppressed miR-145 expression when exposed to Dox. The miR-145 inhibitor reduced the cellular viability of 3T3-L1 cells. Taken together, these data suggest that miR-145 regulates CtsD-mediated cell death in adipocytes. These findings may have valuable implications concerning the molecular mechanism of CtsD-mediated cell death in obesity, suggesting that CtsD could be a useful therapeutic tool for the prevention and treatment of obesity by regulating fat cell numbers.