• Title/Summary/Keyword: Pit-1 gene

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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.

Effects of Chromium (VI) Exposure on the Placental Function and Reproduction in Rats (6가 크롬 폭로가 랫트의 태반 기능과 출산에 미치는 영향)

  • Lee, Heun;Moon, Deog-Hwan;Lee, Chae-Un;Kang, Sung-Goo;Son, Byung-Chul;Kim, Dae-Hwan;Lee, Chang-Hee;Kim, Jung-Won;Lee, Chae-Kwan;Chun, Jin-Ho
    • Journal of Preventive Medicine and Public Health
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    • v.37 no.2
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    • pp.157-165
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    • 2004
  • Objectives : This study aimed to investigate the toxic effects of chromium (VI) on the placental function and reproduction in rats. For the study, the placental prolactin-growth hormone (PRL-GH) gene expression, placental trophoblast cell differentiation and reproductive data were analyzed. Methods : The pregnancies of F344 Fisher rats were checked by the presence of a copulatory plug or sperm in the vaginal smear, which was defined as day 0 of the pregnancy. Pregnant rats were divided into the three groups. The control group was given tap water (chromium level < 0.001 ppm) and the remaining groups were given 250 or 750 ppm of chromium (VI) [as potassium dichromate], from day 7 to 19 of the pregnancy. Rats were sacrificed at days 11 and 20 of pregnancy. The mRNA levels of PRL-GH and Pit-1a and b isotype genes were analyzed by Northern blot hybridization and reverse transcription-polymerase chain reaction (RT-PCR). The hormonal concentration was analyzed by radioimmunoassay, and the differentiation of placental trophoblast cells were observed by histochemical studies. Reproductive data, such as placental and fetal weights, pregnancy period, and litter size, were surveyed at day 20 of pregnancy and after birth. A statistical analysis was carried out using the SAS program (version 8.1). Results : The mRNA levels of the prolactin-growth hormone (PRL-GH) family of genes were dose dependently reduced by chromium exposure. The mRNA levels of Pit-1a and b isotype genes that induce the expression of the PRL-GH family of genes were also reduced by chromium exposure. The PRL-GH hormonal concentration in the rat placenta, fetus and maternal blood were decreased by chromium exposure. In the middle stage of pregnancy (day 11), a high dose of chromium suppressed the differentiation of spongiotrophoblast cells that secret the PRLGH hormones. In the last stage of pregnancy (day 20), a high dose of chromium induced apoptosis of placental cells. Reproductive data, such as placental and fetal weights, litter size, were reduced, but the pregnancy period was extended in the group exposed to chromium compared with the controls. Conclusion : Chromium (VI) disrupts the ordered functions of the placenta, which leads to reproductive disorders in rats.

Human Periodontal Ligament Fibroblasts Support the Osteoclastogenesis of RAW264.7 Cells (치주인대섬유아세포가 파골세포분화에 미치는 영향)

  • Lee, Ho;Jeon, Yong-Seon;Choi, Seoung-Hwan;Kim, Hyung-Seop;Oh, Kwi-Ok
    • Journal of Periodontal and Implant Science
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    • v.32 no.4
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    • pp.733-744
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    • 2002
  • The fibroblasts are the principal cells in the periodontal ligament of peridontium. As the periodontal ligament fibroblasts (PDLF) show similar phenotype with osteoblasts, the PDLF are thought to play an important role in alveolar bone remodeling. Cell-to-cell contacted signaling is crucial for osteoclast formation. Recently it has been reported that PDLJ enhance the bone resorbing activity of osteoclasts differentiated from hematopoietic preosteoclasts. The aims of this study were to $clarify\;^{1)}$ the mechanism of PDLF-induced osteoclastogenesis $and\;^{2)}$ whether we can use preosteoclast cell line instead of primary hematopoietic preosteoclast cells for studying the mechanism of PDLF-induced osteoclastogenesis. Osteoclastic differentiation of mouse macrophage cell line RAW264.7 was compared with that of mouse bone marrow-derived M-CSF dependent cell (MDBM), a well-known hematopoietic preosteoclast model, by examining, 1) osteoclast-specific gene expression such as calcitonin receptor, M-CSF receptor (c-fms), cathepsin K, receptoractivator nuclear factor kappa B (RANK) ,2) generation of TRAP(+) multinucleated cells (MNCs), and 3) generation of resorption pit on the $OAAS^{TM}$ plate. RAW264.7 cultured in the medium containing of soluble osteoclast differentiation Factor (sODF) showed similar phenotype with MDBM-derived osteoclasts, those are mRNA expression pattern of osteoclast-specific genes, TRAP(+) MNCs generation, and bone resorbing abivity. Formation of resorption pits by osteoclastic MNCs differentiated from sODF-treated RAW264.7, was completely blocked by the addition of osteoprotegerin (OPG), a soluble decoy receptor for ODF, to the sODF-containing culture me야um. The effects of PDLF on differentiation of RAW264.7 into the TRAP(+) multinucleated osteoclast-like cells were examined using coculture system. PDLF were fxed with paraformaldehyde, followed by coculture with RAW264.7, which induced formation of TRAP(+) MNCs in the absence of additional treatment of sODF. When compared with untreated and fixed PDLF (fPDLF), IL-1 ${\beta}$-treated, or lipopolysaccha-ride-treated and then fixed PDLF showed two-folld increase in the supporting activity of osteoclastogenesis from RAW264.7 coculture system. There were no TRAP(+) MNCs formation in coculture system of RAW264.7 with PDLF of no fixation. These findigs suggested that we can replace the primary hematopoietic preosteoclasts for RAW264. 7 cell line for studying the mechanism of PDLF-induced osteoclastogenesis, and we hypothesize that PDLF control osteoclastogenesis through ODF expression which might be enhanced by inflammatory signals.

Effect of Banhahoobak-tang (Banxiahoupo-tang) Extract (BHTe) on Psychological Stress (반하후박탕(半夏厚朴湯)이 생쥐의 심리적 스트레스에 미치는 영향과 유전자 분석)

  • Choi, Geum-Ae;Cho, Su-In;Kim, Kyeong-Su;Choi, Chang-Won;Wei, Tung-Shuen;Yang, Seung-Joung;Park, Soo-Yeon;Kim, Kyeong-Ok
    • Journal of Oriental Neuropsychiatry
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    • v.26 no.2
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    • pp.117-130
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    • 2015
  • Objectives: Banhahoobak-tang has been used to treat plum-pit qi, chest and hypochondriac distension, moist or greasy tongue coat, and wiry slow or wiry slippery pulse. It might be used to control coughing and vomiting. We observed that Banhahoobak-tang extract (BHTe) had anti-psychological stress effect. The objective of this study was to determine the effect of BHTe on restoring the transcriptional regulation of genes related to psychological stress. Methods: After giving psychological stress to mice, BHTe was orally administered at 100 mg/kg/day for five days. After extracting whole brain tissue from the mice, the gene expression changes were determined by microarray. Transcription factor binding site (TFBS) analysis showed up- and down-regulated genes related to psychological stress were protected by BHTe and segregated according to the structure of TFBS. We performed text based Pubmed search to select significant target genes involved in psychological stress affected by BHTe. Results: 1. Serum corticosterone level was decreased in the BHTe administered group, although the psychological stress was increased. 2. The BHTe administered group had no significant change in noradrenaline content in brain tissue, but the psychological stress group had decreased level. 3. The BHTe administered group had increased time of staying at open-arm than the psychological stress group. 4. Microarray revealed that TANK and RARA genes were up-regulated genes while AES, CDC42, FOS, NCL, and PVR were down-regulated genes by psychological stress but restored by BHTe.