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ERK is involved in the Dkk-1 and TNF-α crosstalk of murine pre-osteoblast MC3T3E1 under mechanical stress

ERK is involved in the Dkk-1 and TNF-α crosstalk of murine ...

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中华口腔医学会第九次全科口腔医学学术会议

作者： Yeke Wu Chengdu University of Traditional Chinese Medicine

Objectives To explore the involvement of ERK signaling in the crosstalk of Dkk-1 and TNF-α, as well as the role of them in various behaviors of murine pre-osteoblast MC3 T3 E1 under compressive and/or tensile *** and Methods i) Forcel four-point bending system was utilized to apply cyclic uniaxial tensile/compressive strain(2,000 μstrain, 0.5 Hz) on MC3 T3 E1 cells, for 0, 1, 3, 6, 12, 24 h, respectively. At each point, cell proliferation, apoptosis, cycle distribution, β-catenin localization, ALP activity, Cyclin D1, Runx2, Osterix, Dkk-1, Wnt5 a, RANKL, OPG and TNF-α mRNA transcriptions were determined, and RANKL/OPG expression ratio was calculated;ii) MC3 T3 E1 were transfected with Dkk-1 silent and overexpression plasmids under tensile/compressive stress for 3 h or 12 h, and subject to the abovementioned measurements, and p-ERK/ERK expression ratio was also measured;iii) on the basis of the abovementioned model, exogenous TNF-α was added and Dkk-1 expression was measured;iv) exogenous TNF-α was added to MC3 T3 E1 with Dkk-1 silent or overexpressed under compressive stress for 3 h or 12 h or 24 h, and Cyclin D1, Wnt5 a, RANKL/OPG, p-ERK/ERK expression(ratio) were measured;v) on the basis of abovementioned model(without exogenous TNF-α), PD98059 was added to block ERK signaling, and TNF-α mRNA was detected;vi) exogenous TNF-α was added to MC3 T3 E1 under compressive stress for 3 h, and Dkk-1 mRNA was detected. Results i) under the tensile stress, cell proliferation, S%, PI and ALP activity of MC3 T3 E1 were significantly up-regulated, β-cateinin was activated for nuclear translocation, while the apoptosis was suppressed. Dkk-1, TNF-α, RANKL/OPG expression(ratio) were obviously diminished, and Cyclin D1,Wnt5 a, Runx2, Osterix, OPG expression were significantly augmented. On the contrary, the indicators of compressive stress group were just the reverse;ii) under tensile/compressive strain, cell proliferation, S%, PI, ALP activity, Wnt5 a、Runx2、Osterix,

关键词： stress pre-osteoblast Dkk-1 TNF-α ERK

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Wnt5a induced bone destruction through activating JNK in pre-osteoblasts and MSCs of collagen-induced arthritis arthritic mice

Wnt5a induced bone destruction through activating JNK in pre...

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第十一届全国免疫学学术大会

作者： Niu menglin Tong Yulong Du Yuxuan Yuan Huihui Zhao Wenming Department of Immunology School of Basic Medical SciencesCapital Medical University Department of Blood Transfusion Peking University Cancer Hospital & Institute

Background:This study aimed to detect the expression of Wnt5 a in the hind paws bone joint tissue of collagen-induced arthritis(CIA) mice and to evaluate the effects and mechanisms of Wnt5 a on pre-osteoblastic cells(MC3T3-E1 cells) and bone mesenchymal stem cells(MSCs) in secretion and *** and Results:In vivo,Wnt5 a was overexpression in the hind paw joints upon CIA in *** immunofluorescent staining showed that the high level expression of Wnt5 a was mainly localized in osteoblasts from the CIA mice compared to normal controls,which relating to the increasing osteoblasts *** vitro,compared with Wnt3 a treated group,adding Wnt5 a reduced the expression of OPG and increased the IL-6 level in MC3T3-E1 cells and MSCs,while no significantly changes were determined in the expression of IL-1β,TNF-α and *** addition,Wnt5 a treatment induced the MC3T3-E1 cells activation and expression of alkaline phosphatase(ALP) and Runt-related transcription factor 2(RUNX2) in both mR NA and ***,the Ror2 mR NA expression was also higher in Wnt5 a treated cells than ***,Wnt5 a up-regulated JNK signal protein expression and decreased significantly β-*** results demonstrated that the up-regulating of Wnt5 a in osteoblasts caused synovitis,bone erosion and abnormal new bone formation through changing the levels of OPG,IL-6,ALP and ***,Wnt5 a possible bound its receptor Ror2,stimulated the JNK signal pathway and suppressed the canonical Wnt pathway to aggravate arthritis in CIA ***:The study suggested that Wnt5 a closely contributed to the pathogenesis of rheumatoid arthritis(RA).

关键词： Wnt5a CIA pre-osteoblast Secretion Differentiation RA

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