• Title/Summary/Keyword: Cathepsin

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Proteomic Analysis of Bovine Longissimus Muscle Satellite Cells during Adipogenic Differentiation

  • Rajesh, Ramanna Valmiki;Park, Mi-Rim;Heo, Kang-Nyeong;Yoon, Du-Hak;Kim, Tae-Hun;Lee, Hyun-Jeong
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.5
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    • pp.685-695
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    • 2011
  • Satellite cells are skeletal muscle progenitor/stem cells that reside between the basal lamina and plasma membranes of skeletal fibers in vivo. These cells can give rise to both myogenic and adipogenic cells. Given the possible role for differentiation of satellite cells into adipocytes in marbling and in some pathological disorders like sarcopenia, knowledge of the proteins involved in such process remains obscure. Using two-dimensional polyacrylamide gel electrophoresis coupled with mass spectrometry, we investigated the proteins that are differentially expressed during adipogenic differentiation of satellite cells from bovine longissimus muscle. Our proteome mapping strategy to identify the differentially expressed intracellular proteins during adipogenic differentiation revealed a total of 25 different proteins. The proteins up-regulated during adipogenic differentiation of satellite cells like Cathepsin H precursor, Retinal dehydrogenase 1, Enoyl-CoA hydratase, Ubiquinol-cytochrome-c reductase, T-complex protein 1 subunit beta and ATP synthase D chain were found to be associated with lipid metabolism. The down-regulated proteins like LIM protein, annexin proteins, cofilin-1, Rho GDP-dissociation inhibitor 1 and septin-2, identified in the present study were found to be associated with myogenesis. These results clearly demonstrate that the adipogenic conversion of muscle satellite cells is associated with the up-regulated and down-regulated proteins involved in adipogenesis and myogenesis respectively.

Identification of Differentially Expressed Genes in Human Small Cell Lung Carcinoma Using Subtractive Hybridization

  • Ahn Seung-Ju;Choi Jae-Kyoung;Joo Young Mi;Lee Min-A;Choi Pyung-Rak;Lee Yeong-Mi;Kim Myong-Shin;Kim So-Young;Jeon Eun-Hee;Min Byung-In;Kim Chong-Rak
    • Biomedical Science Letters
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    • v.10 no.3
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    • pp.195-202
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    • 2004
  • Lung cancer is a leading cause of cancer death worldwide; however, despite major advances in cancer treatment during the past two decades, the prognostic outcome of lung cancer patients has improved only minimally. This is largely due to the inadequacy of the traditional screening approach of diagnosis in lung cancer, which detects only well­established overt cancers and fails to identify precursor lesions in premalignant conditions of the bronchial tree. In recent years this situation has fundamentally changed with the identification of molecular abnormalities characteristic of premalignant changes; these concern tumour suppressor genes, loss of heterozygosity at crucial sites and activation of oncogenes. Basic knowledge at the molecular level has extremely important clinical implications with regard to early diagnosis, risk assessment and prevention, and therapeutic targets. In this study we used a 'cap-finder' subtractive hybridization method, 'long distance' polymerase chain reaction (PCR), streptavidin magnetic beads mediated subtraction, and spin column chromatography to detect differential expression genes of human small cell lung carcinoma. We have now isolated ninety two genes that expressed differentially in the human small cell lung carcinoma cells and analyzed of 12 clones with sequencing, nine cDNAs include tapasin (NGS-17) mRNA, BC200 alpha scRNA, chromosome 12q24 PAC RPCI3-462E2, protein phosphatase 1 (PPPICA), translocation protein 1 (TLOC1), ribosomal protein S24 (RPS24) mRNA, protein phosphatase (PPEF2), cathepsin Z, MDM2 gene and three novel genes. They may be oncogenesis­related proteins.

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Purification and Characterization of a 25 kDa Cathepsin L-like Protease from the Hemocyte of Coleopteran Insect, Tenebrio molitor Larvae

  • Jang, Kyung-Suk;Cho, Mi-Young;Choi, Hye-Won;Lee, Kang-Moon;Kim, Mi-Hee;Lee, Young-Un;Kurata, Shoichiro;Natori, Shunji;Lee, Bok-Luel
    • 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.

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Inhibitory Effects of Boesenbergia pandurata on Age-Related Periodontal Inflammation and Alveolar Bone Loss in Fischer 344 Rats

  • Kim, Haebom;Kim, Changhee;Kim, Do Un;Chung, Hee Chul;Hwang, Jae-Kwan
    • Journal of Microbiology and Biotechnology
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    • v.28 no.3
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    • pp.357-366
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    • 2018
  • Periodontitis, an infective disease caused by oral pathogens and the intrinsic aging process, results in the destruction of periodontal tissues and the loss of alveolar bone. This study investigated whether Boesenbergia pandurata extract (BPE) standardized with panduratin A exerted anti-periodontitis effects, using an aging model representative of naturally occurring periodontitis. In aged rats, the oral administration of BPE ($200mg{\cdot}kg^{-1}{\cdot}day^{-1}$) for 8 weeks significantly reduced the mRNA and protein expression of $interleukin-1{\beta}$, nuclear factor-kappa B, matrix metalloproteinase (MMP)-2, and MMP-8 in gingival tissues (p < 0.01). In alveolar bone, histological analysis with staining and micro-computed tomography revealed the attenuation of alveolar bone resorption in the BPE-treated aged group, which led to a significant reduction in the mRNA and protein expression of nuclear factor of activated T-cells c1 (NFATc1), c-Fos, tartrate-resistant acid phosphatase, and cathepsin K (p < 0.01). BPE not only increased the expression of osteoblast differentiation markers, such as alkaline phosphate, and collagen type I (COL1A1), but also increased the ratio of osteoprotegerin to RANKL. Collectively, the results strongly suggested that BPE is a natural resource for the prevention or treatment of periodontal diseases.

Inhibitory Effects of Panduratin A on Periodontitis-Induced Inflammation and Osteoclastogenesis through Inhibition of MAPK Pathways In Vitro

  • Kim, Haebom;Kim, Mi-Bo;Kim, Changhee;Hwang, Jae-Kwan
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.190-198
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    • 2018
  • Periodontitis is an inflammatory disease caused by microbial lipopolysaccharide (LPS), destroying gingival tissues and alveolar bone in the periodontium. In the present study, we evaluated the anti-inflammatory and anti-osteoclastic effects of panduratin A, a chalcone compound isolated from Boesenbergia pandurata, in human gingival fibroblast-1 (HGF-1) and RAW 264.7 cells. Treatment of panduratin A to LPS-stimulated HGF-1 significantly reduced the expression of interleukin-$1{\beta}$ and nuclear factor-kappa B (NF-${\kappa}B$), subsequently leading to the inhibition of matrix metalloproteinase-2 (MMP-2) and MMP-8 compared with that in the LPS control ($^{**}p$ < 0.01). These anti-inflammatory responses were mediated by suppressing the mitogen-activated protein kinase (MAPK) signaling and activator protein-1 complex formation pathways. Moreover, receptor activator of NF-${\kappa}B$ ligand (RANKL)-stimulated RAW 264.7 cells treated with panduratin A showed significant inhibition of osteoclastic transcription factors such as nuclear factor of activated T-cells c1 and c-Fos as well as osteoclastic enzymes such as tartrate-resistant acid phosphatase and cathepsin K compared with those in the RANKL control ($^{**}p$ < 0.01). Similar to HGF-1, panduratin A suppressed osteoclastogenesis by controlling MAPK signaling pathways. Taken together, these results suggest that panduratin A could be a potential candidate for development as a natural anti-periodontitis agent.

Cloned Placenta of Korean Native Calves Died Suddenly at Two Months after Birth Displays Differential Protein Expression

  • Kim Hong Rye;Kang Jae Ku;Lee Hye Ran;Yoon Jong Taek;Seong Hwan Hoo;Jung Jin Kwan;Park Chang Sik;Jin Dong Il
    • Reproductive and Developmental Biology
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    • v.29 no.2
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    • pp.63-68
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    • 2005
  • Cloned calves derived from somatic cell nuclear transfer (SCNT) have been frequently lost by sudden death at 1 to 3 month following healthy birth. To address whether placental anomalies are responsible for the sudden death of cloned calves, we compared protein patterns of 2 placentae derived from SCNT of Korean Native calves died suddenly at two months after birth and those of 2 normal placentae obtained from AI fetuses. Placental proteins were separated using 2-Dimensional gel electrophoresis. Approximately 800 spots were detected in placental 2-D gel stained with coomassie-blue. Then, image analysis of Malanie III (Swiss Institute for Bioinformatics) was performed to detect variations in protein spots between normal and SCNT placentae. In the comparison of normal and SCNT samples, 8 spots were identified to be up-regulated proteins and 24 spots to be down-regulated proteins in SCNT placentae, among which proteins were high mobility group protein HMG1, apolipoprotein A-1 precursor, bactenecin 1, tropomyosin beta chain, $H^+-transporting$ ATPase, carbonic anhydrase II, peroxiredoxin 2, tyrosine-rich acidic matrix protein, serum albumin precursor and cathepsin D. These results suggested that the sudden death of cloned calves might be related to abnormal protein expression in placenta.

Suppression of Inflammation, Osteoclastogenesis and Bone Loss by PZRAS Extract

  • Li, Liang;Park, Young-Ran;Shrestha, Saroj Kumar;Cho, Hyoung-Kwon;Soh, Yunjo
    • Journal of Microbiology and Biotechnology
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    • v.30 no.10
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    • pp.1543-1551
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    • 2020
  • Panax ginseng has a wide range of activities including a neuroprotective effect, skin protective effects, enhanced DNA repairing, anti-diabetic activity, and protective effects against vascular inflammation. In the present study, we sought to discover the inhibitory effects of a mixture of natural products containing Panax ginseng, Ziziphus jujube, Rubi fructus, Artemisiae asiaticae and Scutellaria baicalensis (PZRAS) on osteoclastogenesis and bone remodeling, as neither the effects of a mixture containing Panax ginseng extract, nor its molecular mechanism on bone inflammation, have been clarified yet. PZRAS upregulated the levels of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GSH-R) and glutathione peroxidase (GSH-Px) and reduced malondialdehyde (MDA) in LPS-treated RAW264.7 cells. Moreover, treatment with PZRAS decreased the production of IL-1β and TNF-α. PZRAS also inhibited osteoclast differentiation through inhibiting osteoclastspecific genes like MMP-2, 9, cathepsin K, and TRAP in RANKL-treated RAW264.7 cells. Additionally, PZRAS has inhibitory functions on the RANKL-stimulated activation of ERK and JNK, which lead to a decrease in the expression of NFATc1 and c-Fos. In an in vivo study, bone resorption induced by LPS was recovered by treatment with PZRAS in bone volume per tissue volume (BV/TV) compared to control. Furthermore, the ratio of eroded bone surface of femurs was significantly increased in LPS-treated mice compared to vehicle group, but this ratio was significantly reversed in PZRAS-treated mice. These results suggest that PZRAS could prevent or treat disorders with abnormal bone loss.

The Root from Heracleum moellendorffii Exerts Anti-Inflammatory Activity via the Inhibition of NF-κB and MAPK Signaling Activation in LPS-Stimulated RAW264.7 Cells

  • Park, Su Bin;Kim, Ha Na;Kim, Jeong Dong;Park, Gwang Hun;Son, Ho-Jun;Eo, Hyun Ji;Song, Jeong Ho;Jeong, Hyung Jin;Jeong, Jin Boo
    • 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.

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Effect of Atractylodis Rhizoma Alba on Osteoclast Formation (백출의 파골세포 분화에 미치는 영향)

  • Park, Sung-Tae;Lee, Myeung-Su;Jeon, Byung-Hun;Park, Kie-In;Oh, Jae-Min
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.1
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    • pp.109-114
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    • 2011
  • Atractylodis Rhizoma Alba is commonly used herbal medicine and it has been known that has immuno-regualtory effects and anti-cancer effects. The inhibition of osteoclastogenesis is essential for the prevention and treatment of osteoporosis. The aim of this study was to evaluate the effects of Atractylodis Rhizoma Alba on osteoclast differentiation in vitro and on resorbing activity of osteoclast. Osteoclast formation was evaluated in bone marrow cells (BMC) in the presence or absence of Atractylodis Rhizoma Alba. The expression of c-fos, tartrate-resistant acid phosphatase (TRAP), OSCAR, DC-STAMP, cathepsin K, MafB and NFATc1 mRNA in osteoclast precursor were assessed by RT-PCR. The levels of TNF receptor-associated factor-6 (TRAF-6), c-fos and NFATc1 protein were assessed by Western blot analysis. Also the correlation with MAPKs and NF-${\kappa}B$ pathways were measured by using Western blot analysis. With bone resorption study, I tried to evaluate the inhibitory effects of Atractylodis Rhizoma Alba on mature osteoclast function. Atractylodis Rhizoma Alba inhibited the RANKL induced osteoclastic differentiation from bone marrow macrophage in a dose dependant manner without cellular toxicity. Gene expression of c-fos and NFATc1 was significantly down regulated with Atractylodis Rhizoma Alba treatment. Atractylodis Rhizoma Alba markedly inhibited the RANKL-induced osteoclastogenesis through suppression of nuclear factor kappa b (NF-${\kappa}B$) pathway, down stream pathway of p38, ERK and JNK pathway. Taken together, I concluded that Atractylodis Rhizoma Alba have beneficial effect on osteoporosis by inhibition of osteoclast differentiation and by inhibition of functioning osteoclast. Thus I expect that Atractylodis Rhizoma Alba could be a treatment option for osteoporosis.

Inhibitory Effects of Rhizoma Arisaematis on Osteoclast Differentiation and Bone Resorption (파골세포의 분화와 뼈 흡수에 천남성의 억제 효과)

  • Lee, Myeung-Su;Lee, Chang-Hoon;Park, Kie-In;Kim, Ha-Young
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.1
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    • pp.65-70
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    • 2011
  • Osteoclasts play a critical role in bone-related diseases such as osteoporosis and rheumatoid arthritis by resorbing the bone. Recently, natural products from plants have been extensively studied as therapeutic drugs to treat and prevent various diseases. Here, we examined the effects of rhizoma arisaematis on ostoclast differentiation and bone resorption. We showed that rhizoma arisaematis significantly suppressed receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL)-induced osteoclast differentiation in bone marrow-derived macrophages (BMMs) in a dose dependent manner but have little or no effect on the cytotoxicity of BMMs and RAW264.7 cells. We found that rhizoma arisaematis iarrow-ed the RANKL-induced c-Fos and nuclear factor of activated T cells (NFAT)c1, which is a master regulator of osteoclast differentiation. Furthermore, rhizoma arisaematis suppressed the mRNA expression of tartrate resistant-acid phosphatase and cathepsin K iaduced by RANKL in BMMs. in y chanistic studies, rhizoma arisaematis considerably iarrow-ed I-${\kappa}B$ degradation, which is a negative regulator of NF-${\kappa}B$, but iaduced the phosphderlation of p-38, ERK, and JNK.MMlso, we found that rhizoma arisaematis significantly iarrow-ed osteoclastic bone resorption. Taken tarether, our results suggest that rhizoma arisaematis suppresses osteoclast differentiation through down-regulatd the mRANKL-induced c-Fos and NFATc1 expression and iarrow-s bone resorption.