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목적: Receptor activator of nuclear factor-kB ligand (RANKL)과 osteoprotegerin (OPG)는 조골세포에서 생성되는 사이토카인인데, RANKL은 파골세포 형성을 촉진시키고 OPG는 RANKL의 decoy 수용체로 작용함으로써 파골세포 형성을 억제한다. Peroxisome proliferation-activated receptor gamma (PPARγ)는 지방세포 분화에 중요한 역할을 하는 것으로 알려져 있지만 이것은 골 재형성에도 관여한다. 따라서 본 연구에서는 PPARγ 작용제인 rosiglitazone이 조골세포에서 RANKL과 OPG 발현을 어떻게 조절하는지 조사했다.
방법: MC3T3-E1 조골세포를 여러 농도의 rosiglitazone으로 24시간 처리하여 세포생존능을 MTT 방법으로 측정했다. PPARγ 유전자의 발현은 통상적인 PCR로 관찰했으며, RANKL과 OPG mRNA와 단백질의 발현은 real-time RT-PCR과 Western blot으로 각각 정량 분석했다.
결과: MC3T3-E1 조골세포에 존재하는 PPARγ 유전자 발현은 rosiglitazone 0.5μM 처리 시 대조군과 유사했으나 5μM 처리 시 현저히 증가했다. 세포생존능의 경우 rosiglitazone 1~10μM 범위에서 대조군과 차이를 보이지 않았다. Rosiglitazone (5μM) 처리에 의해 OPG mRNA의 발현은 대조군에 비해 2.3배 증가하였고 RANKL mRNA의 경우 1/5로 감소했다. 그러나 이러한 변화는 PPARγ 길항제인 GW9662 처리에 의해 모두 대조군 수준으로 회복되었다. Rosiglitazone에 의한 OPG와 RANKL 단백질 발현 또한 이들의 mRNA 발현 변화와 유사한 경향을 보였다.
결론: 조골세포에서 rosiglitazone에 의해 PPARγ가 활성화됨으로써 RANKL 발현은 감소되고 동시에 OPG 발현은 증가되는데, 이는 파골세포 형성을 억제할 것으로 사료된다.
Objectives: Osteoblasts secrete receptor activator of nuclear factor-kB ligand (RANKL) and osteoprotegerin (OPG). RANKL stimulates osteoclastogenesis but OPG inhibits it by binding with RANKL. Although peroxisome proliferation-activated receptor gamma (PPARγ) has a major role in adipocyte differentiation, it is also involved in bone remodeling. The aim of this study was to examine whether rosiglitazone, the PPARγ agonist, regulates the expression of RANKL and OPG in osteoblasts.
Methods: MC3T3-E1 osteoblasts were cultured and treated with different concentrations of rosiglitazone. Cell viability was assayed with MTT. A conventional PCR was used for the detection of PPARγ gene. Expression of RANKL and OPG mRNA and their protein levels was analyzed by quantitative real-time RT- PCR and Western blot, respectively.
Results: PPARγ mRNA was expressed in MC3T3-E1 osteoblasts and its expression was greatly increased by treatment of 5μM rosiglitazone. Treatment with 1 to 10μM rosiglitazone did not affect cell viability. Compared with vehicle, rosiglitazone (5μM) up-regulated OPG mRNA expression by 2.3-fold and down- regulated RANKL mRNA expression by 5-fold, which was reversed by the PPARγ antagonist GW9662. The level of RANKL and OPG protein varied as compared to their mRNA expression.
Conclusion: These results demonstrate that activation of PPARγ by rosiglitazone induces both down- regulation of RANKL and up-regulation of OPG in osteoblasts, which is likely to cause an inhibition of osteoclastogenesis.| 번호 | 참고문헌 | 국회도서관 소장유무 |
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| 1 | OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis. ![]() |
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| 2 | PPARgamma insufficiency enhances osteogenesis through osteoblast formation from bone marrow progenitors. ![]() |
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| 3 | osteoprotegerin-defident mice develop early onset osteoporosis and arterial calcification Genes Dev 1998;12(9): 1260-8. | 미소장 |
| 4 | Influence of Glucocorticoids on Human Osteoclast Generation and Activity ![]() |
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| 5 | PTH Differentially Regulates Expression of RANKL and OPG ![]() |
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| 6 | Effects of arachidonic acid, docosahexaenoic acid and prostaglandin E 2 on cell proliferation and morphology of MG-63 and MC3T3-E1 osteoblast-like cells ![]() |
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| 7 | IL-6, leukemia inhibitory factor, and oncostatin M stimulate bone resorption and regulate the expression of receptor activator of NF-kappa B ligand, osteoprotegerin, and receptor activator of NF-kappa B in mouse calvariae. ![]() |
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| 8 | Transforming growth factor-beta stimulates the production of osteoprotegerin/osteoclastogenesis inhibitory factor by bone marrow stromal cells. ![]() |
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| 9 | Nitric oxide regulates receptor activator of nuclear factor-kappaB ligand and osteoprotegerin expression in bone marrow stromal cells. ![]() |
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| 10 | Rosiglitazone causes bone loss in mice by suppressing osteoblast differentiation and bone formation. ![]() |
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| 11 | Bone is a target for the antidiabetic compound rosiglitazone. ![]() |
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| 12 | Enhanced bone formation in lipodystrophic PPARγhyp/hyp mice relocates haematopoiesis to the spleen ![]() |
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| 13 | Expression of Endothelin-1 and Its Receptors in Cisplatin-Induced Acute Renal Failure in Mice | 소장 |
| 14 | Peroxisome proliferator-activated receptor activators modulate the osteoblastic maturation of MC3T3-E1 preosteoblasts ![]() |
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| 15 | Association of Bone Mineral Density with a Polymorphism of the Peroxisome Proliferator-Activated Receptor � Gene: PPAR� Expression in Osteoblasts ![]() |
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| 16 | Activation of Peroxisome Proliferator-activated Receptor-gamma Inhibits the Runx2-mediated Transcription of Osteocalcin in Osteoblasts ![]() |
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| 17 | Osteotropic agents regulate the expression of osteoclast differentiation factor and osteoprotegerin in osteoblastic stromal cells. ![]() |
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| 18 | The expression of osteoprotegerin and RANK ligand and the support of osteoclast formation by stromal-osteoblast lineage cells is developmentally regulated. ![]() |
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| 19 | Activation of peroxisome proliferator-activated receptor-gamma pathway inhibits osteoclast differentiation ![]() |
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| 20 | Stimulation of resorption in cultured mouse calvarial bones by thiazolidinediones. ![]() |
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| 21 | Thiazolidinediones inhibit osteoclast-like cell formation and bone resorption in vitro. ![]() |
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| 22 | Different skeletal effects of the peroxisome proliferator activated receptor (PPAR)α agonist fenofibrate and the PPARγ agonist pioglitazone ![]() |
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| 23 | A novel PPARg agonist, KR62776, suppresses RANKL-induced osteoclast differentiation and activity by inhibiting MAP kinase pathways ![]() |
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| 24 | Activation of peroxisome proliferator–activated receptor-γ inhibits differentiation of preosteoblasts ![]() |
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| 25 | TZDs and Bone: A Review of the Recent Clinical Evidence ![]() |
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