• Title/Summary/Keyword: c-fos mRNA and protein

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Orostachys japonicus Hexane Fraction Attenuates Pro-inflammatory Cytokines in LPS-activated Macrophage Cells by Suppression of AP-1 and IRF3 Transcription Factors (LPS로 유도된 대식세포에 대한 와송 핵산추출물의 AP-1과 IRF3 전사인자의 억제에 의한 전염증성 사이토카인의 감소 효과)

  • Lee, Hyeong-Seon
    • Microbiology and Biotechnology Letters
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    • v.48 no.3
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    • pp.310-315
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    • 2020
  • Orostachys japonicus (O. japonicus) is known as a medicinal plant for the treatment of various symptoms. This study investigated the anti-inflammatory effect of the hexane fraction from O. japonicus (OJH) on the LPS-stimulated response in RAW 264.7 macrophage cells. This study was conducted to confirm the effect of cell cytotoxicity and production of reactive oxygen species (ROS) in OJH-treated macrophage cells. Additionally, pro-inflammatory cytokines and transcription factors were determined using RT-PCR and western blotting assay. OJH showed no change in lactate dehydrogenase (LDH) levels and exhibited reduced ROS levels in LPS-induced inflammatory cells. Moreover, OJH significantly suppressed the mRNA levels of proinflammatory cytokines, including IL-1β, IL-2, IL-6, TNF-α, and IP-10. Furthermore, OJH effectively inhibited the protein levels of AP-1 (p-c-Jun and p-c-Fos) and p-IRF3 in a dose-dependent manner. In conclusion, our results demonstrate that OJH exhibits strong anti-inflammatory activities via regulation of inflammatory factors.

Suppressive Effects of Cyanidin-3-glucoside on Th2 Cytokines Production in RBL-2H3 Cells (RBL-2H3 세포에서 Cyanidin-3-glucoside의 Th2 사이토카인 발현 억제 효과)

  • Jeong, Hwa Hyun;Yoon, Soo Jeong;Pyo, Myoung Yun
    • YAKHAK HOEJI
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    • v.57 no.5
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    • pp.309-315
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    • 2013
  • Cyanidin-3-glucoside (C3G), an anthocyanin, is one of the most widespread dietary flavonoids. We investigated the effects of C3G in PMA/ionomycin (PI)-induced RBL-2H3 cells. C3G inhibited the production of IL-4 and IL-13 and also decreased the level of mRNA in a dose-dependent manner. Furthermore, western blot analysis implied that C3G down-regulated the protein level of c-Jun, NF-ATc1 and NF-${\kappa}B$ but not c-Fos. Taken together, we suggest that C3G may have suppressive effects on Th2 cytokines and will be studied further to develop as functional foods that help alleviate allergy symptoms.

The Effect of Ampelopsis japonica (Thunb.) Makino on Osteoclastogenesis and Expression of Osteoclast-Related Gene (백렴(白蘞)의 파골세포 분화 및 관련 유전자 발현 억제에 미치는 영향)

  • Hongsik Kim;Sumin Lee;Minsun Kim;Jae-Hyun Kim;Yejin Kang;Seoung Jun Kwon;Youngwoo Nam;Seungwoo Yoo;Hong-Seok Choi;SeonJin Huh;Youngjoo Sohn;Hyuk-Sang Jung
    • The Korea Journal of Herbology
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    • v.38 no.5
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    • pp.9-19
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    • 2023
  • Objectives : Osteoporosis is a systemic skeletal disorder characterized by reduced bone mineral density and increased risk of fractures. Bisphosphonates and selective estrogen receptors, which are bone resorption inhibitors that are currently widely used as osteoporosis treatments, show serious side effects when administered for a long time. Research on bone resorption inhibitors that complement the problems of existing treatments is needed. The purpose of this study was to investigate the effect of inhibiting osteoclast differentiation and activity on the tuberous root of Ampelopsis japonica (Thunb.) Makino (AM). Methods : After extracting AM using distilled water and ethanol, the inhibitory effects of the two solvents on osteoclast differentiation were compared using the RANKL-induced in vitro experimental model and the TRAP assay kit. The impact of AM on bone resorption was investigated through the pit formation assay, and its effect on F-actin formation was assessed through fluorescent staining. Additionally, protein and mRNA expression levels of osteoclast differentiation markers (NFATc1, c-Fos, TRAP and ATP6v0d2) and resorption markers (MMP-9, CTK, and CA2) were analyzed via western blot and RT-PCR. Results : AM treatment significantly decreased the number of TRAP-positive cells and pit formation area. Furthermore, AM suppressed both the protein and mRNA expression of NFATc1 and c-Fos, key transcription factors involved in osteoclast differentiation and it downregulated the expression of osteoclast-associated genes such as TRAP, CTK, MMP-9, CA2, and ATP6v0d2. Conclusions : These results suggest that AM can inhibit bone resorption and osteoclast differentiation, indicating its potential for use in the treatment and prevention of osteoporosis.

Effect of Water Extract of Schisandra Chinensis on Osteoclast Differentiation (오미자 물 추출물이 파골세포 분화에 미치는 영향)

  • Lee, Yan;Lee, Ho-Sub;Jang, Sung-Jo;Song, Jeong-Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.24 no.5
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    • pp.848-853
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    • 2010
  • Bone maintains its homeostasis through balance between bone resorbing osteoclasts and bone forming osteoblasts. Thus, unusual balance between osteoclasts and osteoblasts leads to pathological bone diseases, such as osteoporosis, rheumatoid arthritis, autoimmune arthritis, periodontitis. Schisandra chinensis well known traditional herbal has been used for treatment of diseases in China, Korea, Japan, andothers. Recently, research studies have demonstrated that the lignans found in Schisandra chinensis stimulate osteoblasts and suggest that it may be helpful against osteoporosis. However, the inhibitory effect of water extract of Schisandra chinensis on osteoclast differentiation remains largely unknown. In this study, Water extract of Schisandra chinensis markedly suppressed RANKL-induced osteoclast differentiation in cultures of BMMs without cytotoxicity. The mRNA expression of c-Fos, NFATc1, and TRAP induced by RANKL was inhibited by water extract of Schisandra chinensis. It also suppressed c-Fos and NFATc1 protein expression. Taken together, these results suggest that water extract of Schisandra chinensis has the potential to serve as a treatment of bone disease such as osteoporosis.

Diphlorethohydroxycarmalol Suppresses Ultraviolet B-Induced Matrix Metalloproteinases via Inhibition of JNK and ERK Signaling in Human Keratinocytes

  • Piao, Mei Jing;Kumara, Madduma Hewage Susara Ruwan;Kim, Ki Cheon;Kang, Kyoung Ah;Kang, Hee Kyoung;Lee, Nam Ho;Hyun, Jin Won
    • Biomolecules & Therapeutics
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    • v.23 no.6
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    • pp.557-563
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    • 2015
  • Skin aging is the most readily observable process involved in human aging. Ultraviolet B (UVB) radiation causes photo-oxidation via generation of reactive oxygen species (ROS), thereby damaging the nucleus and cytoplasm of skin cells and ultimately leading to cell death. Recent studies have shown that high levels of solar UVB irradiation induce the synthesis of matrix metalloproteinases (MMPs) in skin fibroblasts, causing photo-aging and tumor progression. The MMP family is involved in the breakdown of extracellular matrix in normal physiological processes such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes such as arthritis and metastasis. We investigated the effect of diphlorethohydroxycarmalol (DPHC) against damage induced by UVB radiation in human skin keratinocytes. In UVB-irradiated cells, DPHC significantly reduced expression of MMP mRNA and protein, as well as activation of MMPs. Furthermore, DPHC reduced phosphorylation of ERK and JNK, which act upstream of c-Fos and c-Jun, respectively; consequently, DPHC inhibited the expression of c-Fos and c-Jun, which are key components of activator protein-1 (AP-1, up-regulator of MMPs). Additionally, DPHC abolished the DNA-binding activity of AP-1, and thereby prevented AP-1-mediated transcriptional activation. These data demonstrate that by inactivating ERK and JNK, DPHC inhibits induction of MMPs triggered by UVB radiation.

Inhibitory Effects of Water Extract of Cervi parvum cornu, Carthami tinctorii fructus and Their Combination on Osteoclast Differentiation and Bone Resorption (녹용(鹿茸), 홍화자(紅花子) 단일 및 혼합 물 추출물( 抽出物)의 파골세포(破骨細胞) 분화(分化) 억제(抑制)와 골흡수(骨吸收) 억제(抑制) 효과(效果))

  • Ann, Ji-Young;Kim, Ju-Ho;Ki, Ji-Ye;Kwak, Han-Bok;Oh, Jae-Min;Kim, Yun-Kyung
    • Herbal Formula Science
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    • v.18 no.2
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    • pp.167-182
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    • 2010
  • Cervi parvum cornu (Deer Antler) and Carthami tinctorii fructus (Also known as Carthami seed) are widely used for treating osteoporosis and rheumatoid arthritis. In this study, We found out that the water extract of Cervi parvum cornu(WECPC), Carthami tinctorii fructus(WECTF) and their combination have effects of suppressing the RANKL-induced osteoclast differentiation. We assayed mRNA expression levels of NFATc1, c-Fos, TRAP and GAPDHS from bone marrow macrophages(BMMs) by means of RT-PCR. Similarly, the protein expression levels of NFATc1, c-Fos, MAPKs and $\beta$-actin in cell lysates were analyzed by means of Western blotting. then we determined the anti-osteoporotic effects of WECPC, WECTF and their combination using Lipopolysaccharide (LPS)-induced bone-loss mouse. WECPC, WECTF and their combination showed remarkable inhibition on RANKL-treated osteoclast differentiation without cytotoxicity. WECPC suppressed degradation of I-${\kappa}B$. WECPC, WECTF and their combination down-regulated the induction of c-Fos and NFATc1 by RANKL. Lastly, in vivo data showed that WECPC, WECTF and their combination rescued the bone erosion by LPS treatment. Thus, these results demonstrate that WECPC, WECTF and their combination can be efficacious remedies for bone-loss diseases such as osteoporosis and rheumatoid arthritis.

Inhibitory Effects of Achyranthis Bidentatae Radix on Osteoclast Differentiation and Bone Resorption (우슬의 파골세포 분화 억제와 골 흡수 억제효과)

  • Kim, Ju-Ho;Ki, Ji-Ye;Ann, Ji-Young;Park, Hye-Jung;Kim, Hyun-Ju;Kwak, Han-Bok;Oh, Jae-Min;Kim, Yun-Kyung
    • The Korea Journal of Herbology
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    • v.25 no.1
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    • pp.65-74
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    • 2010
  • Objectives : Achyranthis Bidentatae Radix (ABR) has been used for treating of many symptoms especially osteoporosis and rheumatoid arthritis. In this study, we determined the effects of water extract of ABR in RANKL (Receptor Activator for Nuclear Factor $\kappa$ B Ligand)-induced osteoclast differentiation culture system. Methods : We assayed mRNA expression levels of NFATc1, c-Fos, TRAP, OSCAR, $FcR{\gamma}$, DAP12 and GAPDH in bone marrow macrophages (BMMs) treated with ABR. The protein expression levels of NFATc1, c-Fos, MAPKs and $\beta$-actin in cell lysates treated with ABR were analysed by Western blotting. In addition we determined the effects of water extract of ABR on LPS-induced bone-loss mouse. Results : Water extract of ABR showed remarkable inhibition on RANKL-treated osteoclast differentiation without cytotoxicity. ABR down-regulated the induction of c-Fos and NFATc1 by RANKL. ABR suppressed phosphorylation of JNK, p38 and I-${\kappa}B$. ABR rescued bone erosion by LPS induction in vivo study. Conclusions : These results demonstrate that ABR may be a useful remedy for curing of bone-loss disease such as osteoporosis.

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.

Forsythiae Fructus Extract Inhibits RANKL-Induced Osteoclast Differentiation and Prevent Bone Loss in OVX-Induced Osteoporosis Rat (연교의 파골세포 분화 및 골 흡수 억제 기전 연구)

  • Eom, Ji-Whan;Kim, Jae-Hyun;Kim, Minsun;Kim, Sangwoo;Shin, Hwajeong;Jung, Hyuk-Sang;Sohn, Youngjoo
    • Korean Journal of Acupuncture
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    • v.36 no.2
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    • pp.115-126
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    • 2019
  • Objectives : Osteoporosis is a condition characterized by low bone mass and increased bone fragility. It has become a major problem of senior citizens. The purpose of this study is to experiment the effect of water extract of Forsythiae Fructus (wFF) on osteoclast differentiation; and the other purpose is to examine the effect of wFF on osteoporosis in ovariectomized rat. Methods : To investigate the effect of wFF on osteoclast differentiation and activity, RAW 264.7 cells were used. The number of TRAP positive cell, TRAP activity, pit area, mRNA expression of makers (RANK, TRAP, CA II, CTK, MMP-9, NFATc1, c-Fos), protein expression of makers (NFATc1, c-Fos) were investigated. For in vivo study, 40 female Sprague-Dawley (SD) rats were induced osteoporosis by ovariectomy (OVX) and then tested for anti-osteoporosis effect by administration of wFF. Results : wFF suppressed osteoclatogenesis, TRAP activity and pit area formation. Moreover, wFF decreased the expression of master differentiation factors (NFATc1, c-Fos) and also reduced the osteoclastogenesis-related markers (TRAP, CA II, CTK, MMP-9). These suggest that wFF inhibit osteoclasts differentiation and bone resorption. In the OVX rat model, wFF inhibited decreasing of BMD and trabecular area. Conclusions : Forsythiae Fructus should be effective for osteoporosis prevention and treatment.

Inhibitory Effects of Ssangbohwan on Osteoclast Differentiation and Bone Resorption (쌍보환 추출물의 파골세포 분화 억제와 골 흡수 억제효과)

  • Kim, Seong Joung;Lee, Jeong Ju;Kim, June Hyun;Jo, So Hyun;Park, Min Cheol;Jo, Eun Heui
    • Journal of Acupuncture Research
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    • v.32 no.3
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    • pp.69-81
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    • 2015
  • Purpose : The first purpose of this study is to find out whether the water extract of Rehmanniae Radix Preparat(RRP), Cuscutae Semen(CS) and their combination(Ssangbohwan, SBH) have the effect of suppressing Receptor activator of nuclear factor kappa-B ligand(RANKL)-induced osteoclast differentiation. The second purpose of this study is to find out whether the water extract of RRP, CS and SBH have the effect of inhibiting osteoporosis in an osteoporosis model induced by lipopolysaccharide(LPS). Methods : After promoting differentiation of osteoclasts by treating the RANKL, we observed the effect by the administration of RRP, CS and SBH. In addition, by means of Reverse transcription polymerase chain reaction(RT-PCR), we assayed mRNA expression levels of NFATc1, c-Fos, TRAP and GAPDHS(Glyceraldehyde-3-phosphate dehydrogenase, spermatogeni) from bone marrow macrophages(BMMs). Similarly, the protein expression levels of NFATc1 (Nuclear factor of activated T-cells, cytoplasmic1), C-Fos, MAPKs(Mitogen-activated protein kinases) and ${\beta}$-actin in cell lysates were analyzed by means of Western Blotting. Finally, we determined the anti-osteoporotic effects of RRP, CS and SBH, through the use of Lipopolysaccharide-induced bone-loss mouse. Results : RRP, CS and SBH showed remarkable inhibitive effect on RANKL-treated osteoclast differentiation without cytotoxicity. SBH inhibited the phosphorylation of p38, Jun N-terminal kinases(JNK), and I-${\kappa}B$ and down-regulated the induction of c-Fos and NFATc1 by RANKL. RRP, CS suppressed degradation of I-${\kappa}B$, but it did not affect c-Fos and NFATc1 by RANKL. Lastly, in vivo data showed that RRP and SBH prevented bone erosion by LPS treatment. Conclusions : These results demonstrate SBH can be effective remedy for bone-loss diseases such as osteoporosis.