• 동의생리병리학회지
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• 제25권1호
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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.

• Biomolecules & Therapeutics
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• 제28권4호
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• pp.337-343
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• 2020

Activation of osteoclast and inactivation of osteoblast result in loss of bone mass with bone resorption, leading to the pathological progression of osteoporosis. The receptor activator of NF-κB ligand (RANKL) is a member of the TNF superfamily, and is a key mediator of osteoclast differentiation. A flavanone glycoside isolated from the fruit of Poncirus trifoliata, poncirin has anti-allergic, hypocholesterolemic, anti-inflammatory and anti-platelet activities. The present study investigates the effect of poncirin on osteoclast differentiation of RANKL-stimulated RAW264.7 cells. We observed reduced formation of RANKL-stimulated TRAP-positive multinucleated cells (a morphological feature of osteoclasts) after poncirin exposure. Real-time qPCR analysis showed suppression of the RANKL-mediated induction of key osteoclastogenic molecules such as NFATc1, TRAP, c-Fos, MMP9 and cathepsin K after poncirin treatment. Poncirin also inhibited the RANKL-mediated activation of NF-κB and, notably, JNK, without changes in ERK and p38 expression in RAW264.7 cells. Furthermore, we assessed the in vivo efficacy of poncirin in the lipopolysaccharide (LPS)-induced bone erosion model. Evaluating the micro-CT of femurs revealed that bone erosion in poncirin treated mice was markedly attenuated. Our results indicate that poncirin exerts anti-osteoclastic effects in vitro and in vivo by suppressing osteoclast differentiation. We believe that poncirin is a promising candidate for inflammatory bone loss therapeutics.

• Journal of Microbiology and Biotechnology
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• 제28권2호
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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.

• Journal of Microbiology and Biotechnology
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• 제30권10호
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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.

• 동의생리병리학회지
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• 제25권1호
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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.

• Molecules and Cells
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• 제40권10호
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• pp.752-760
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• 2017

A2B adenosine receptor (A2BAR) is known to be the regulator of bone homeostasis, but its regulatory mechanisms in osteoclast formation are less well-defined. Here, we demonstrate the effect of A2BAR stimulation on osteoclast differentiation and activity by RANKL. A2BAR was expressed in bone marrow-derived monocyte/macrophage (BMM) and RANKL increased A2BAR expression during osteoclastogenesis. A2BAR stimulation with its specific agonist BAY 60-6583 was sufficient to inhibit the activation of ERK1/2, p38 MAP kinases and $NF-{\kappa}B$ by RANKL as well as it abrogated cell-cell fusion in the late stage of osteoclast differentiation. Stimulation of A2BAR suppressed the expression of osteoclast marker genes, such as c-Fos, TRAP, Cathepsin-K and NFATc1, induced by RANKL, and transcriptional activity of NFATc1 was also inhibited by stimulation of A2BAR. A2BAR stimulation caused a notable reduction in the expression of Atp6v0d2 and DC-STAMP related to cell-cell fusion of osteoclasts. Especially, a decrease in bone resorption activity through suppression of actin ring formation by A2BAR stimulation was observed. Taken together, these results suggest that A2BAR stimulation inhibits the activation of ERK1/2, p38 and $NF-{\kappa}B$ by RANKL, which suppresses the induction of osteoclast marker genes, thus contributing to the decrease in osteoclast cell-cell fusion and bone resorption activity.

• Journal of Microbiology and Biotechnology
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• 제28권3호
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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.

• Journal of Microbiology and Biotechnology
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• 제27권2호
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• pp.372-379
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• 2017

The transport of lysosomal enzymes into the lysosomes depends on the phosphorylation of their chains and the binding of the phosphorylated residues to mannose-6-phosphate receptors. The efficiency of separation depends more on the phosphodiesterases (PDEs) than on the activity of the phosphorylation of mannose residues and can be determined in vitro. PDEs play important roles in regulation of the activation of lysosomes. The expression of proteins was confirmed by western blotting. All PDE4 series protein expression was reduced in high concentrations of rolipram. As a result of observing the fluorescence intensity after rolipram treatment, the lysosomal enzyme was activated at low concentrations and suppressed at high concentrations. High concentrations of rolipram recovered the original function. Antimicrobial activity was not shown in either 10 or $100{\mu}M$ concentrations of rolipram in treated HeLa cells in vitro. However, the higher anticancer activity at lower rolipram concentration was shown in lysosomal enzyme treated with $10{\mu}M$ of rolipram. The anticancer activity was confirmed through cathepsin B and D assay. Tranfection allowed examination of the relationship between PDE4 and lysosomal activity in more detail. Protein expression was confirmed to be reduced. Fluorescence intensity showed decreased activity of lysosomes and ROS in cells transfected with the antisense sequences of PDE4 A, B, C, and D. PDE4A showed anticancer activity, whereas lysosome from cells transfected with the antisense sequences of PDE4 B, C, and D had decreased anticancer activity. These results showed the PDE4 A, B, C, and D are conjunctly related with lysosomal activity.

• 대한약리학회지
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• 제18권2호
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• pp.15-25
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• 1982

한국(韓國)에서 빈발(頻發)하는 뇌졸중증(腦卒中症) 환자(患者)의 혈장(血漿)에서 혈전형성(血栓形成)에 관계되는 인자(因子)들을 조사하고, in vitro로 sodium niroprusside의 혈소판(血少板)응집에 대(對)한 작용(作用)을 관찰(觀察)하여 뇌졸중증(腦卒中症) 응급시(應急時) 치료적응용(治療的應用)의 가능성(可能性)을 검토(檢討)하였다. 1) 뇌졸중증(脚卒中症) 환자(患者)의 혈장내(血漿內) 미세혈전형성(微細血栓形成)은 건강인(健康人)에 비(比)하여 유의(有意)하게 (p<0.001) 높았다. 2) 뇌졸중증(腦卒中症) 환자(患者)의 혈장내(血漿內) 과산화지질(過酸化脂質) 및 cathepsin D치(値)는 건강인(健康人)에서 보다 현저히 증가되어 있었다. 3) 건강인(健康人)의 혈소판(血少板)은 1 및 $2{\mu}M\;ADP$에 대(對)하여 2차반응(次反應)에서는 응집률(凝集率)이 감소(減少)된데 반(反)하여 뇌졸중증환자(腦卒中症患者)에서는 응집률(凝集率)이 항진(亢進)되었다. 4) $K^+-free\;PSS$, ouatain, 13-HPLA 및 Cd으로 전처치(前處置)한 소 뇌저동맥(腦底動脈), 토끼 대동맥(大動脈) 및 사람 제대동맥(臍帶動脈)은 혈소판응집률(血少板凝集率)을 증가(增加)시켰다. 그중 $K^+-free$에 처치(處置)한 것이 가장 강(强)하게 작용(作用)하였다. 5) Sodium nitropusside로 전처치(前處置)하였을 때 $K^+-free\;PSS$에 처치(處置)된 혈관(血管)에 의한 항응집반응(向凝集反應)은 용량(用量)에 의존(依存)하여 현저(顯著)히 억제(抑制)되었으며 hydralazine이나 diazoxide에 의하여는 아무런 작용(作用)을 받지 아니하였다. 6) Dibutyryl cyclic AMP 및 8-bromo cyclic GMP 모두 혈소판응집률(血小板凝集率)을 감소시켰고, 후자(後者)는 전자(前者)에 비(比)하여 월등(越等)히 현저(顯著)하였다. sodium nitroprusside에 의한 항응집률(抗凝集率)은 methylene blue 전처치(前處置)에 의하여 길항(拮抗)되었으나, bovine hemoglobin전처치에 의하여는 영향(影響)을 받지 아니하였다. 이상(以上)의 성적(成績)을 종합(綜合)하면, 뇌졸중증(腦卒中症)때, 특히 뇌혈전증(腦血栓症)의 응급치료시(應急治療時) sodium nitroprusside의 응용(應用)이 가능(可能)하다고 사료(思料)되며, 이에 대(對)하여 임상적(臨床的) 치료(治療)가 기대되는 바이다.

• Journal of Acupuncture Research
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• 제22권3호
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• pp.13-22
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• 2005

손상된 조직의 보호와 항염증작용이 있는 것으로 알려진 유근피를 수액추출하여 골다공증의 치료에 응용 가능한가를 실험하였다. 이전의 실험에서 유근피 추출액이 파골세포를 함유한 장골세포의 치료를 통해 Cathepsin K를 억제하는 것을 확인 하였었다. 이를 통하여 유근피는 골다공증 치료에서 골재흡수억제제로 서의 prodrug의 역할을 할 수 있음을 시사하였다. 본 실험에서는 MC3T3-El pre-osteoblastic 세포조직을 이용하여 골화를 유발한 상태에서 유근피가 골아세포의 성장과 감작에 대한 것을 in vitro로 연구하였다. 이 결과 유근피는 용량과 시간의존적으로 ALP의 활동을 향상시킴으로써 강화작용이 있음을확인할 수 있었다. 투여량은 최소치 $50{\mu}g/m{\ell}$에서 최대치 $150{\mu}g/m{\ell}$에서 관찰되었다. $100{\mu}g/m{\ell}$ UD에서 bone morphogenetic protein-2의 향상을 관찰할 수 있었으며, MC3T3-El 세포내의 ALP mRNA농도역시 증가 하였다. $60{\mu}g/m{\ell}$ UD에서 Type I collagen mRNA에 대해서 오랜 배양 기간 동안 약간의 증가를 나타내었으나 15-20일 사이의 배양에서는 급격히 유전자 발현을 억제하는 것으로 나타났다. 이러한 결과는 골아 세포의 감작을 통해 유근피가 골대사에 영향을 미침을 시사하는 것이다. 그러므로 추후 연구를 통하여 일반적인 골대사질환에 유근피를 적극적으로 활용할 수 있음을 알 수 있었다.