tailieunhanh - Báo cáo khoa học: "Effects of osteoprotegerin from transfection of pcDNA3.1(+)/chOPG on bioactivity of chicken osteoclasts"

Tuyển tập các báo cáo nghiên cứu về bệnh học thý y được đăng trên tạp chí Acta Veterinaria Scandinavica cung cấp cho các bạn kiến thức về bệnh thú yđề tài: Effects of osteoprotegerin from transfection of (+)/chOPG on bioactivity of chicken osteoclasts. | Hou et al. Acta Veterinaria Scandinavica 2011 53 21 http content 53 1 21 AVS ACTAVETERINARIA s5jdmovccJa SCANDINAVICA RESEARCH Open Access Effects of osteoprotegerin from transfection of chOPG on bioactivity of chicken osteoclasts Lele Hou Jiafa Hou Jing Yao and Zhenlei Zhou Abstract Background Osteoprotegerin OPG has been reported to prevent bone resorption by inhibiting the formation function and survival of osteoclasts in a variety of animal models. However the effects of OPG on bone metabolism in avian species have not been described. The objective of this study was to investigate the effects of chicken OPG chOPG expressed in chicken embryo fibroblasts CEFs on chicken osteoclast function in vitro. Methods The chOPG sequence containing the open reading frame ORF was amplified from chicken embryo frontal bone and inserted into the vector. chOPG was transiently transfected into CEFs by lipofectamine 2000. Transcription of OPG mRNA and expression of chOPG recombinant protein were detected by reverse transcription polymerase chain reaction RT-PCR and indirect immunofluorescence. The level of chOPG recombinant protein was detected by enzyme-linked immunosorbent assay. The suspension of osteoclasts was separated from chicken embryos and divided into three groups control group group and chOPG group . The percentage of osteoclast apoptosis was detected by flow cytometry. The tartrate-resistant acid phosphatase TRAP secreted by osteoclasts was measured by the diazol method. The resorbing activity of osteoclasts was evaluated by the area of lacunae on bone flaps and the concentration of calcium in the supernatant liquid of osteoclasts. Results 48 h after transfection the exogenous OPG gene transcription was detected by RT-PCR. After 72 h the CEFs transfected from chOPG displayed green fluorescence and the concentration of chOPG protein was ng mL. After chicken osteoclasts were cultured .

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