• Journal of Veterinary Science
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• v.23 no.4
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• pp.47.1-47.11
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• 2022

Background: In lipopolysaccharide-induced RAW264.7 cells, Aster tataricus (AT) inhibits the nuclear factor kappa-light-chain-enhancer of activated B cells and MAPKs pathways and critical pathways of osteoclast development and bone resorption. Objectives: This study examined how aster saponin A2 (AS-A2) isolated from AT affects the processes and function of osteoclastogenesis induced by receptor activator of nuclear factor kappa-B ligand (RANKL) in RAW264.7 cells and bone marrow macrophages (BMMs). Methods: The cell viability, tartrate-resistant acid phosphatase staining, pit formation assay, polymerase chain reaction, and western blot were carried out to determine the effects of AS-A2 on osteoclastogenesis. Results: In RAW264.7 and BMMs, AS-A2 decreased RANKL-initiated osteoclast differentiation in a concentration-dependent manner. In AS-A2-treated cells, the phosphorylation of ERK1/2, JNK, and p38 protein expression were reduced considerably compared to the control cells. In RAW264.7 cells, AS-A2 suppressed the RANKL-induced activation of osteoclast-related genes. During osteoclast differentiation, AS-A2 suppressed the transcriptional and translational expression of NFATc1 and c-Fos. AS-A2 inhibited osteoclast development, reducing the size of the bone resorption pit area. Conclusion: AS-A2 isolated from AT appears to be a viable therapeutic therapy for osteolytic illnesses, such as osteoporosis, Paget's disease, and osteogenesis imperfecta.

• The Korea Journal of Herbology
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• v.29 no.2
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• pp.61-67
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• 2014

Objectives : Aconiti Lateralis Preparata Radix (Aconitum Carmichaeli, AC) and Cinnamomi Cortex (Cinnamomi Cortex, CC) have been treated to elderly for kidney yang enhancement in Korean traditional medicine. In this study, the effects of water extract of AC and CC on RANKL (Receptor Activator for Nuclear Factor ${\kappa}B$ Ligand)-induced osteoclast differentiation were evaluated in culture system. Methods : MTT assay was used to evaluate the potential cytotoxicity of AC and CC extracts in bone macrophage marrows (BMMs) stimulated with M-CSF. TRAP (tartrate-resistant acid phosphatase) staining and TRAP activity were performed to know the inhibitory effect on osteoclast differentiation. The protein expression levels of nuclear factors such as activated T cell(NFAT)c1, c-Fos, MAPKs and ${\beta}$-actin in cell lysates treated with AC and CC extracts were analysed by western blotting. Results : AC, CC extracts and their co-administration inhibited significantly RANKL-induced osteoclast differentiation in BMMs in a dose dependent manner without toxicity. Each AC and CC extracts inhibited the phosphorylation of p38. Also, AC and CC extracts, respectively, inhibited the protein expression of c-Fos and NFATc1 more than Co-administration of AC and CC even if all treatments did. It was observed that RANKL-induced degradation of I-${\kappa}B$ is significantly suppressed by all treatments. Conclusions : Taken together, It was concluded that AC and CC have beneficial effect on osteoporosis by inhibition of osteoclast differentiation. Thus, Atractylodis AC and CC could be a treatment option for osteoporosis.

• Journal of Applied Biological Chemistry
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• v.66
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• pp.7-14
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• 2023

The effects of fermented Achyranthes japonica Nakai, Angelica gigas Nakai, and Eucommia ulmoides Oliver extracts (FAAE) on regulation of inflammation and apoptosis were investigated in primary cultured rat cartilage cells. To identify the protective effects of FAAE against H₂O₂, cell survival was measured by MTT assay. Smad3, Collagen type I, MMP3, and MMP13 were measured by real-timpe PCR and westernbot and the inflammatory (NF-κB pathway, COX-2, iNOS) factors were determined by western blot. The apoptosis related factors (JNK, c-Fos, c-Jun, caspase 3, Bax, and Bcl-2) were determined by western blot. FAAE significantly increased the follwing: H₂O₂ treated cell survival, mRNA and protein expression of Smad 3, collagen type I. In addition, FAAE significantly decreased the protein expression of inflammatory and apoptosis related factors. This study suggests that FAAE have a protection effect of chondrocytes through inhibition of inflammation and apoptosis. Thus, FAAE is a therapeutic potential food componet in osteoarthritis.

• Journal of Periodontal and Implant Science
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• v.41 no.4
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• pp.167-175
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• 2011

Purpose: Periodontal ligament (PDL) cell differentiation into osteoblasts is important in bone formation. Bone formation is a complex biological process and involves several tightly regulated gene expression patterns of bone-related proteins. The expression patterns of bone related proteins are regulated in a temporal manner both in vivo and in vitro. The aim of this study was to observe the gene expression profile in PDL cell proliferation, differentiation, and mineralization in vitro. Methods: PDL cells were grown until confluence, which were then designated as day 0, and nodule formation was induced by the addition of 50 ${\mu}g$/mL ascorbic acid, 10 mM ${\beta}$-glycerophosphate, and 100 nM dexamethasone to the medium. The dishes were stained with Alizarin Red S on days 1, 7, 14, and 21. Real-time polymerase chain reaction was performed for the detection of various genes on days 0, 1, 7, 14, and 21. Results: On day 0 with a confluent monolayer, in the active proliferative stage, c-myc gene expression was observed at its maximal level. On day 7 with a multilayer, alkaline phosphatase, bone morphogenetic protein (BMP)-2, and BMP-4 gene expression had increased and this was followed by maximal expression of osteocalcin on day 14 with the initiation of nodule mineralization. In relationship to apoptosis, c-fos gene expression peaked on day 21 and was characterized by the post-mineralization stage. Here, various genes were regulated in a temporal manner during PDL fibroblast proliferation, extracellular matrix maturation, and mineralization. The gene expression pattern was similar. Conclusions: We can speculate that the gene expression pattern occurs during PDL cell proliferation, differentiation, and mineralization. On the basis of these results, it might be possible to understand the various factors that influence PDL cell proliferation, extracellular matrix maturation, and mineralization with regard to gene expression patterns.

• YAKHAK HOEJI
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• v.56 no.5
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• pp.326-332
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• 2012

Mast cells play pivotal pathologic roles in allergic disease involving T helper 2 (Th2) cytokine such as interleukin (IL)-4 and IL-13. Fisetin has been known as an anti-allergic agent having inhibitory effects on the IL-4 and IL-13 gene expressions in inflammatory immune cells. However, its molecular mechanisms for suppressive effects of fisetin on IL-4 and IL-13 in activated mast cells have been incompletely elucidated. In this study we found that fisetin significantly inhibited the phorbol 12-myristate 13-acetate (PMA) and ionomycin (PI)-induced production of IL-4 and IL-13 in mast cells. The levels of mRNA were dramatically decreased by fisetin, indicating the suppression might be regulated at the transcriptional levels. Western blot analysis of the nuclear expression of various transcription factors involved in the promoter activation indicated that suppression of c-Fos was prominent together with significant down-regulation of nuclear factor of activated T-cell (NF-AT) and NF-${\kappa}B$, but not c-Jun. Furthermore, the nuclear expression of GATA binding protein 2 (GATA-2) transcription factor was significantly down-regulated by fisetin. Taken together, our study indicated fisetin has suppressive effects on IL-4 and IL-13 gene expression through the regulation of selective transcription factors.

• 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.

• Journal of Korean society of Dental Hygiene
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• v.24 no.3
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• pp.219-227
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• 2024

Objectives: The pulp is the center of the tooth containing nerves and blood vessels. The condition in which the pulp becomes inflamed due to caries or periodontitis is called pulpitis. Pulpitis is a difficult-to-treat disease and causes peripheral nerve tissue changes and severe pain; however, the relationship between neuronal activity and voltage-gated sodium channel 1.7 (Nav1.7) expression in the trigeminal ganglion (TG) during pulpitis has not been well studied. In this study, we found that experimentally induced pulpitis activates Nav1.7 expression in the periphery, leading to neuronal overexpression in the TG. Thus, we sought to identify ways to regulate this process. Methods: Acute pulpitis was induced in rat maxillary molars by treating the pulp with allyl isothiocyanate (AITC). Three days later, in vivo optical imaging was used to record and compare neural activities in the TG. Western blotting was used to identify molecular changes in terms of the expression of extracellular signal-regulated kinase (ERK), c-Fos, transient receptor potential ankyrin 1 (TRPA1), and collapsin response mediator protein-2 (CRMP2) in the brain stem. Results: The results confirmed the neurological changes in the TGs of the pulpitis model, and histological and molecular biological evidence confirmed that increased Nav1.7 expression induced by pulpitis leads to pain. Furthermore, selective inhibition of Nav1.7 resulted in changes in neural activity, suggesting that pulpitis induces increased Nav1.7 expression, and that effective control of Nav1.7 could potentially reduce pain. Conclusions: The inhibition of overexpressed Nav1.7 channels may modulate nociceptive signal processing in the brain and effectively control pain associated with pulpitis.

• The Korean Journal of Physiology and Pharmacology
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• v.5 no.2
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• pp.123-131
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• 2001

To investigate the effects of electrical stimulation on vestibular compensation, which is the recovery of vestibular symptoms following unilateral labyrinthectomy (UL), intermittent electrical stimulation was applied to the injured vestibular portion in Sprague-Dawley rats. Vestibuloocular and vestibulospinal reflexes, electrical activity and expression of c-Fos protein in medial vestibular nuclei (MVN) were measured with time following UL. Spontaneous nystagmus occurred with frequency of $2.9{\pm}0.2$ beats/sec at 2 hours after UL and disappeared after 72 hours. Electrical stimulation decreased the frequency of nystagmus significantly till 24 hours after UL. Roll head deviation was $107{\pm}9.7^{\circ}$ at 2 hours after UL and the deviation was maintained till 72 hours, but electrical stimulation decreased the deviation significantly 6 hours after UL. Resting activity of type I neurons in ipsilateral MVN to the injured vestibular side decreased significantly compared with control at 6 and 24 hours after UL, but the activity of type I neurons was recovered to control level by electrical stimulation at 24 hours after UL. Gain of type I neurons induced by sinusoidal rotation of 0.1 Hz decreased significantly till 24 hours after UL, but electrical stimulation restored the activity at 24 hours. The gain of type II neurons decreased significantly at 6 hours after UL, but electrical stimulation restored the activity. Expression of c-Fos protein was asymmetric between bilateral MVN till 24 hours after UL, but the asymmetry disappeared by electrical stimulation 6 hours after UL. These results suggest that electrical stimulation to the injured vestibular portion facilitates vestibular compensation following UL by restoration of symmetry of neuronal activity between bilateral vestibular nuclei resulting from increased activity in ipsilateral vestibular nuclei to the injured side.

• Journal of Acupuncture Research
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• v.34 no.2
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• pp.1-18
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• 2017

Objectives : The purpose of this study was to evaluate the effects of water extracts of Eucommiae cortex (EC), Psoraleae semen (PS), and their combination on receptor activator of nuclear factor-kappa-B ligand (RANKL)-induced osteoclast differentiation. Methods : We assayed the protein expression levels of nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), c-Fos, mitogen-activated protein kinases (MAPKs), and ${\beta}-actin$ in cell lysates using western blotting. Similarly, mRNA expression levels of NFATc1, c-Fos, tartrateresistant acid phosphate (TRAP), and glyceraldehyde-3-phosphate dehydrogenase, spermatogeni (GAPDHS) from bone marrow macrophages (BMMs) were analyzed using reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, we determined the anti-osteoporotic effects of the water extracts of EC, PS, and their combination in a lipopolysaccharide (LPS)-induced bone-loss mouse model. Results : The in vitro data revealed showed that the combination of EC and PS extract showed a more remarkable inhibition of osteoclast differentiation than each herb did alone. The combination downregulated the induction of c-Fos, NFATc1, and TRAP by suppressing the phosphorylation of p38 and c-Jun N-terminal kinases (JNKs) and inhibiting nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$). Lastly, the in vivo data showed that PS reduced the LPS-induced bone erosion. Conclusion : The result of this study suggests that EC and PS could be potential therapeutic agents for bone loss diseases such as osteoporosis.

• Korean Journal of Acupuncture
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• v.21 no.2
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• pp.69-79
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• 2004

Objective : Acupuncture has been used for treatment of numerous disorders, especially for pain control in Oriental Medicine. However, the mechanism of pain control by acupuncture was not clear until now. This study was performed to prove analgesic mechanism of acupuncture treatment at acupoint $ST_{36}$ by observing the changes of abdominal pain and c-Fos expression in the thalamus. Methods : Abdominal pain was induced by acetic acid, and the changes of writhing reflex after acupuncture treatment on $ST_{36}$ and non-acupoints were measured. c-Fos immunohistochemistry was also performed to study the changes of the neuronal activity in the thalamus. Results : The writhing reflex decrease significantly after acupuncturing at $ST_{36}$ compared with control group(p<0.05). The changes of the writhing reflex by non-acupoint acupuncture treatment also showed significant decrease compared with control group(p<0.05). c-Fos expression in the thalamus, especially periventricular part was significantly decreased after acupuncturing as $ST_{36}$ compared with control groups(p<0.05). Conclusion : This study shows that the acupuncture has the analgesic effect in the abdominal pain induced by acetic acid and the thalamus might be a important area for this mechanism.