• Title/Summary/Keyword: Rat bone

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Role of MAP kinase on MMP-13 expression in rat periodontal ligament cells (백서 치주인대세포에서의 MMP-13 mRNA에 대한 MAP kinase의 역할)

  • Chung, Chan-Gil;Cui, De-Zhe;Chung, Hyun-Ju;Kim, Young-Joon
    • Journal of Periodontal and Implant Science
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    • v.36 no.1
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    • pp.85-96
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    • 2006
  • Matrix metalloproteinases (MMPs)는 치주조직내에 존재하는 세포외기질의 유지와 분해에 중요한 역할을 담당하고 있으며 이중 MMP-13은 치주질환의 진행과 깊은 관계가 있다고 알려져 있다. 이번 연구는 치주질환의 진행에 있어서 MMP-13의 활성에 대한 mitogen activated protein(MAP) Kinase의 역할을 구명하기 위해 시행되었다. 백서 치주인대세포에서의 MMP-13 mRNA의 발현은 RT-PCR에 의하여, 그리고 MAP Kinase의 발현은 Western blot에 의하여 측정하였다. $Interleukin-1{\beta}$(IL $-1{\beta}$), Tumor necrosis $factora(TNF-{\alpha})$와 parathyroid hormon(PTH)는 MMP- 13 mRNA 발현을 각각 320%, 180%, 380% 증가시켰으나 bone morphogenetic protein-7(BMP-7)은 MMP-13 mRNA의 발현을 증가시키지 않았다. p38 MAP Kinase 억제제인 SB203580은 IL $-1{\beta}$ 유도 MMP-13의 발현을 약 40% 정도 억제시켰으나, PTH-유도 MMP-13 mRNA의 발현은 억제하지 못했다. IL $-1{\beta}$는 MMP- 13 mRNA의 반감기를 약 2시간 정도로 증가시켰으나, p38 MAP Kinase 억제제로 전처치한 경우에는 반감기가 60분으로 줄어들었다. $IL-1{\beta}$는 p38 MAP kinase와 JNK의 인산화 활성을 증가시켰으나 PTH, $TNF-{\alpha}$와 BMP-7은 p38, JNK, ERK의 활성을 증가시키지 못했다. 이상의 연구결과는 p38 MAP Kinase가 백서 치주인대세포에서의 MMP-13 mRNA 발현을 조절하는데 중요한 역할을 담당함을 시사하였다.

RANKL expression is mediated by p38 MAPK in rat periodontal ligament cells (백서 치주인대세포의 RANKL 발현에 대한 p38 MAPK의 역할)

  • Kim, Chong-Cheol;Kim, Young-Joon;Chung, Hyun-Ju;Kim, Ok-Su
    • Journal of Periodontal and Implant Science
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    • v.34 no.3
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    • pp.489-498
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    • 2004
  • Recent studies have demonstrated that human periodontal ligament cells express receptor activation of nuclear factor ${\kappa}B$ ligand (RANKL) which enhances the bone resorbing activity of osteoclasts differentiated from hematopoietic preosteoclasts. The purpose of this study is to determine the effects of p38 MAPK and JNK kinase upon regulating RANKL and OPG in response to $IL-1{\beta}$(l ng/ml) in rat periodontal ligament cells. Soluble RANKL was measured by immunoassay. The effects of p38 MAPK on RANKL and OPG expression was determined by RT-PCR. The results were as follows: 1. Periodontal ligament cells which stimulated by $IL-1{\beta}$ increased soluble RANKL synthesis by dose-dependent pattern. 2. p38 MAP kinase inhibitor (SB203580) showed regulation of soluble RANKL expression by dose-dependent manners. 3. p38 MAP kinase inhibitor (SB203580) regulated the expression of RANKL, but it dose regulate the expresseion of OPG. 4. JNK (c-jun $NH_2-terminal$ kinase) inhibitor (PD98059) did not regulate mRANKL and mOPG. These results suggested that p38 MAPK play a significant role in RANKL gene expression.

Effect of Ulmus davidiana Planch (Ulmaceae) on T-lymphocyte-producing cytokines such as IL-2, IL-6, and $IFN-{\gamma}$ production in collagen-induced arthritis of rats

  • Kang, Han-Ju;Kim, Kyung-Ho;Jo, Hyun-Seog;Hwang, Min-Seob;Kim, Kap-Sung
    • Journal of Acupuncture Research
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    • v.22 no.2
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    • pp.33-41
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    • 2005
  • Objective : The effect of Ulmus davidiana Planch(UD), which has long been known to have anti-inflammation and protective effects on damaged tissue, inflammation and bone among other functions, on the development of type II collagen (CII)-induced arthritis (CIA) in rats was studied. Methods : Male rats were immunized with an emulsion of $200\;{\mu}g$ of CII and complete Freund's adjuvant (CFA). The rats were then given intraperitoneal stimulation of Ulmus davidiana Planch herbal acupuncture(UDHA)or saline during the experiment. When compared with rats treated with saline as control, UDHA at doses of more than $20{\mu}g/100\;g$ rat once a day for 7 days inhibited the ability of inguinal lymph node cells to produce T cell cytokines interleukin-2, interleukin-6, $IFN-{\gamma}$ when the cells were obtained from rats 14 days after immunization and cultured in vitro with CII. Results : When rats were injected intraperitoneally, UD -treated group and control group rats did not differ significantly when low doses of UD was given to rats. Conclusion : The recommended dose of UD in the management and treatment of rat CIA will be more than $20{\mu}g/100\;g$, which is two-firth of human therapeutic dose. From the results, it was concluded that the effect of UDHA is dependent of dosage.

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Regenerative Effects of Alloplastic Grafts in Rat Periodontal Fenestration Defects (백서 치조골 천공결손부에서의 합성골 이식재의 재생효과)

  • Gang, Yun-Kyung;Park, Joon_Bong;Kwon, Young-Hyuk;Herr, Yeek
    • Journal of Periodontal and Implant Science
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    • v.31 no.2
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    • pp.317-332
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    • 2001
  • The purpose of this study was to investigate effects of the natural coral(NC) and the hydroxyapatite/calcium sulfate hemihydrate(HA/CS) on an early stages of wound healing in the rat periodontal fenestration defects. In this experiment, twelve male rats(Mean : 520g in BW) aged 8 to 9 months were used. Experimental periodontal fenestration defects were surgically created with tapered fissure bur at the buccal surface of the left mandibular 1st, 2nd molars. The buccal aspects of molar roots were carefully denuded of their periodontal ligament through a bony window created in the left mandibles of rats under general anesthesia. Each experimental periodontal fenestration defect was grafted with natural coral and HA/CS, randomly. An area without bone graft was assigned for negative control group. At 10,35 days, rats were serially sacrificed via intracardiac perfusion with 2.5% glutaraldehyde and specimens were processed with Hematoxylin-Eosin stain for light microscopic evaluation. The results of this study were as follows : 1. The defect areas were filled with dense connective tissues at 10 days in control group. But in the test(NC, HA/CS)groups, the connective tissues around graft materials were formed more loosely and the response of inflammation by graft materials itself was not found. 2. The defect areas were filled with new osteoid tissues and new cementum was not formed on the cut root surface at 35 days in the control group. 3. New osteoid tissue formation was more prominent at 35 days in control than test groups. 4. The NC and HA/CS particles were encapsulated by loose connective tissues at 10 days and by dense connective tissues at 35 days, respectively. 5. In the test groups, resorption of graft particles was not found through the experimental time. From the above results, natural coral and hydroxyapatite/calcium sulfate hemihydrate may be biocompatible and osteoconductive and have a weak adverse reaction to the periodontal tissues.

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Hematological Effect of Water Extracts of Cham-Dang-Gui on Cyclophosphamide Induced Anemic Rat (Cyclophosphamide로 유도된 빈혈 흰쥐에서 참당귀 열수추출물이 혈액학적 빈혈지표에 미치는 영향)

  • 한진아;장기효;강순아;조여원
    • Journal of Nutrition and Health
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    • v.36 no.10
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    • pp.1013-1021
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    • 2003
  • Anemia, the condition of the diminished concentration of hemoglobin per erythrocyte is common in patients with cancer and is a frequent complication of myelosuppressive chemotherapy. Cham-Dang-Gui (Angelicae Gigantis Radix) has been used in traditional Korean medicine to treat hematologic deficiencies. In this study, Cyclophosphamide (CYP), an alkylating agent that has a broad spectrum of anti-cancer activities, was intraperitoneally injected into the experimental animals to suppress the bone marrow thereafter, causing anemia. The hemopoietic effects of Cham-Dang-Gui were examined using anemic rat model. Rats were divided into five groups: CON (control group), ANS (CYP-injected + normal diet), AND (CYP-injected + normal diet + Cham-Dang-Gui), ALS (CYP-injected + low iron diet), and ALD (CYP-injected + low iron diet + Cham- Dang-Gui) groups. CYP (30 mg/kg) was intraperitoneally injected to rats for 3 days to induce anemic condition. Saline or Cham-Dang-Gui was administrated orally during the entire experimental period. CYP injection decreased body weight gain and food consumption in comparison with CON group. Oral administration of Cham-Dang-Gui extract with normal iron diet significantly prevented the lower body weight gain. The blood level of hemoglobin, iron status (serum iron, transferrin, ferritin and TIBC) and blood level of vitamin B-12 in Cham-Dang-Gui treated groups were significantly higher than those of Cham-Dang-Gui untreated groups regardless of amount of iron in the diet. Taken together, it could be concluded that the Cham-Dang-Gui extract could improve anemic condition induced by CYP injection by improving hematological value, iron status and vitamin B12 status in rats.

Effects of Yeonsan-Ogye Egg on MIA-induced Osteoarthritis Rat (오계란(烏鷄卵)이 MIA 골관절염 병태 모델에 미치는 영향)

  • Joo, In-Hwan;Kim, Dong-Hee
    • The Korea Journal of Herbology
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    • v.32 no.6
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    • pp.63-69
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    • 2017
  • Objectives : The purpose of this study is to investigate the preventive and therapeutic efficacy of osteoarthritis using a Yeonsan-Ogye egg. so, we researched at effects of Yeonsan-Ogye egg extract on MIA-induced Osteoarthritis animal models. Methods : Yeonsan-Ogye egg extract was administered 500 mg/kg/day, 1000 mg/kg/day and 2000 mg/kg/day to SD-Rat for 2 weeks. After that, osteoarthritis was induced with $60mg/m{\ell}$ of monosodium iodoacetate (MIA) and futher administration was continued for 4 weeks. 3D imaging of cartilage patella were obtained using a Micro-CT system and the pathology change of knee was observed by H&E and safranin-O staining. The weight bearing ratio was measured by incapacitance test meter. MMP-2, MMP-9, COMP, CTX II, calcitonin and glycosaminoglycan level in serum were measured using a ELISA. Results : Micro-CT and Histopathological analysis showed the volume of the patella cartilage and the proteoglycan contents were increased in all groups. also weight bearing ratio was decreased in all groups compared with control group. Calcitonin production was increased in and 2000 mg/kg/day group and glycosaminoglycan production was increased in all groups. In addition, MMP-2, MMP-9, COMP and CTX II production were decreased in 1000 and 2000 mg/kg/day groups respectively in comparison with control. Conclusions : The results for Yeonsan-Ogye egg showed prevention and treatment efficacy against arthritis at serum and the cartilage. These results may be used a remedy for new korea medicine to ease the symptoms mentioned above. also, suggest that Yeonsan-Ogye egg can be used preventive and therapeutic material for osteoarthritis.

The effects of a Mixture of Puerariae Flos Extract and Citri Unshius Pericarpium Extract on Estrogenic Activities and Osteoclastogenesis (갈화(葛花)와 진피(陳皮) 추출물로 이루어진 복합물의 에스트로겐 활성과 파골세포 분화억제효과)

  • Cho, Hosong;Lee, Boyoung;Lee, Won Kyung;Lee, Jun Ho;Park, Dongjun;Choi, Changil;Jin, Mu Hyun;Roh, Seok-Seon;Ju, Young-Sung
    • The Korea Journal of Herbology
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    • v.35 no.3
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    • pp.1-8
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    • 2020
  • Objectives : In this study, we examined the estrogenic activities and anti-osteo clastogenesis effects of PCE17, a mixture of PE (an extract of Puerariae Flos), and CE (an extract of Citri Unshius Pericarpium). Methods : The estrogenic effect of PCE17, PE and CE were examined by ER-β/ERE reporter gene assay and proliferation assay in 293 T and MCF-7 cells. The expression of estrogen-responsive gene and protein were checked by Real Time-PCR (RT-PCR) and Western blotting in MCF-7 cells. Inhibitory effect of PCE17, PE and CE on RANKL-induced osteoclast differentiation were evaluated by TRAP staining and RT-PCR in primary osteoclast precursors from rat bone marrow cells. Results : PCE17 and PE bind to ERs (estrogen receptors) and show estrogenic activities in 293T cells. They also stimulated the proliferation of MCF-7 cells and increased the expression of ER response gene, pS2. Tectorigenin, an active ingredient of PE, shows similar estrogenic activities in MCF-7 cells. PCE17 and CE inhibited RANKL-induced osteoclastogenesis in rat primary osteoclast precursor cells and down-regulated the osteoclast-specific genes of Nfatc1, Ctsk, and Acp5. Conclusions : In conclusion, PCE17 may have therapeutic potential in cases of menopause and osteoporosis.

THE ASPECT OF PROLIFERATION AND BONE NODULE FORMATION IN OSTEOBLAST-LIKE CELLS DERIVED FROM FETAL RAT CALVARIA IN VITRO (백서 태자 두 개관에서 유래된 조골세포의 증식 및 골결절 형성양상)

  • Kim, Shi-Hyeong;Nam, Soon-Hyeun;Shin, Hong-In
    • Journal of the korean academy of Pediatric Dentistry
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    • v.24 no.1
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    • pp.1-17
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    • 1997
  • The purpose of this study was to investigate the aspects of proliferation and bone nodule formation of osteogenic precursor cells. To determine the effects of ascorbic acid and dexamethasone upon capacity of osteoblast proliferation and bone nodule formation, cells were maintained in the presence of one or some of these additives for up to 30 days. Group I culture was maintained in standard medium(DMEM plus 10% plus antibiotics), group II was maintained in supplemented medium containing dexamethasone, group III was maintained in supplemented medium containing ascorbic acid and sodium-${\beta}$-glycerophosphate, and group IV was maintained in supplemented containing ascorbic acid, sodium-${\beta}$-glycerophosphate and dexamethasone. Morphology of bone nodules was observed with light microscope and electron microscope. The results were as follows: ${\bullet}$ Proliferation capacity of osteoblasts was not affected by single use of dexamethasone, but it was chiefly affected by ascorbic acid. ${\bullet}$ Cellular morphology was fibroblastic appearance initially, but, it was gradually changed to polygonal shape accompanied by confluency stage. ${\bullet}$ Pluripotent mesenchymal cells existed during primary culture, they were differentiated to adipocyte, chondrocyte, osteocyte according to culture condition. ${\bullet}$ Dexamethasone increased bone nodule formation under the condition that the culture was maintained with supplemented medium ascorbic acid and sodium-${\beta}$-glycerophosphate. ${\bullet}$ when the cultures were stained with alizarin red, the group supplemented with dexamethasone, ascorbic acid and sodium-${\beta}$-glycerophosphate showed the marked increase of bone nodule formation, but the group supplemented with ascorbic acid and sodium-${\beta}$-glycerophosphate revealed only small amounts of bone nodules. And the groups cultured without ascorbic acid showed no observed any of bone-like mass independent of dexamethasone addition.

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Laminin-1 Expression in Bone Marrow Stromal Cells of Cyclophosphamide-treated Rat (Cyclophosphamide가 흰쥐 골수의 기질세포에서 Laminin-1의 발현에 미치는 영향)

  • Lee, Chang-Hun;Chung, Ho-Sam;Paik, Doo-Jin;Hwang, Se-Jin;Kim, Won-Kyu;Youn, Jee-Hee;Kim, Chong-Kwan
    • Applied Microscopy
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    • v.32 no.4
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    • pp.385-398
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    • 2002
  • The purpose of the present study is to investigate whether stromal cells supporting specific microenvironment for hematopoiesis of bone marrow are affected by toxicants and therapeutic drugs such as antibiotics and anticancer drugs and whether laminin-1 is associated with such effects. SD rats were intraperitoneally injected with 75 mg/kg of cyclophosphamide which is widely used to treat infant's solid tumor, leukemia and myeloma and sacrificed after 3 days, 1 week, 3 weeks or 5 weeks of injection. The bone marrow extracted and paraffin-sectioned was analyzed using immunohistochemical staining. A part of tissues was subjected to electron microscopy following reaction with rabbit anti-laminin antibody, biotinylated goat anti-rabbit IgG conjugated with 12 nm gold particles, and staining with uranyl acetate. 1. The bone marrow tissue at day 3 post injection with cyclophosphamide displayed dilated venous sinus, partial necrotic death, and decreased number of hematopoietic cells. Laminin-1 was intensively stained in the reticular and adipose tissues. 2. Up to 5 weeks post injection, laminin-1 was stained at a low level in the stromal tissue of bone marrow and the number of hematopoietic cell was increased. 3. Deposition of the gold particle which represents laminin-1 expression was observed at the highest level in the stromal cells of bone marrow obtained 3 days after injection, and decreased after 1 to 5 weeks. These results suggest that stromal cells which play a role in supporting microenvironment for bone marrow hematopoiesis augment induction of laminin-1 expression and activation upon administration of cyclophosphamide.

Effect of Agaricus blazei β-Glucan and Egg Shell Calcium Complex on Bone Metabolism in Ovariectomized Rats (난소절제 흰쥐에서 신령버섯의 β-Glucan과 난각 Ca 복합체가 골 대사에 미치는 효과)

  • Noh, Kyung-Hee;Jang, Ji-Hyun;Kim, Jin-Ju;Park, Cherl-Woo;Kim, Jeong-Ok;Kim, Jae-Cherl;Song, Young-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.10
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    • pp.1363-1370
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    • 2006
  • This study was designed to evaluate the effect of Agaricus blazei $\beta-glucan$ and egg shell calcium complex on bone metabolism in ovariectomized (OVX) rats. Forty Sprague-Dewley female rats, 10 weeks of age $(248{\pm}1.7g)$, were divided into 4 groups and fed on the experimental diets for 6 weeks: sham operated control treated with normal diet containing 0.5% calcium (Sham-C), OVX-control treated with normal diet containing 0.5% calcium (OVX-C), $OVX-\beta-glucan$ group treated with $\beta-glucan$ diet containing 0.5% calcium (OVX-G), and $OVX-\beta-glucan$ egg shell calcium complex treated with $OVX-\beta-glucan$ egg shell calcium complex containing 0.5% calcium (OVX-GE). Bone weight of femur was higher in the OVX-GE group than in the other OVX groups. Bone mineral density of femur was significantly different (p<0.05) among the experimental groups and showed the highest level in the OVX-GE group. Calcium absorption rate and retention were higher in the $\beta-glucan$ supplement groups than in the other groups (p<0.05). Alkaline phosphatase activities and osteocalcin levels of serum showed lower in the $\beta-glucan$ supplement groups than in the OVX-C group. Deoxypyridinoline crosslink values of urine, indicator of bone absorption, showed the lowest in the OVX-GE group. The $\beta-glucan$ supplemented groups had a lower bone resorption ratio than in the OVX-C group. We concluded that bioavailability of calcium is higher in $\beta-glucan$ supplement groups compared to those in OVX rats. From the above results, these findings suggest the possibility of using $\beta-glucan$ egg shell calcium complex as a functional food material related to bone metabolism, even though there is no significant difference between the groups of $\beta-glucan$ and $\beta-glucan-egg$ shell calcium complex supplementation.