CAJ | 학술논문

背景:在骨组织中,护骨素的主要作用是抑制破骨细胞的形成和活性,核转录因子κB受体活化素配体的主要作用为刺激破骨细胞的分化和活性,抑制破骨细胞的凋亡.护骨素/核转录因子κB受体活化素配体在破骨功能调控中起重要作用.近来研究发现脂联素促进成骨细胞增殖和分化,脂联素与骨代谢相互偶联.但脂联素对破骨细胞的作用不明.目的:观察脂联素对成骨细胞核转录因子κB受体活化素配体和护骨素表达的影响,进一步分析其对破骨细胞生成的作用.设计、时间及地点:对比观察实验,于2007-0612008-12在中南大学湘雅二医院代谢内分泌研究所实验室完成.材料:外科手术取正常成人髂前上棘松质骨用于细胞培养,临床标本由湘雅医院提供.方法:分别用0,3,10和30 mg/L脂联素干预人成骨细胞48 h,检测护骨素和核转录因子κB受体活化素配体mRNA与蛋白表达.并用脂联素干预成骨细胞与外周血单核细胞共同培养系统,观察其对破骨细胞生成的影响.主要观察指标:分别采用荧光定量聚合酶链反应和酶联免疫吸附法检测人成骨细胞护骨素和核转录因子κB受体活化素配体mRNA与蛋白表达.抗酒石酸磷酸酶染色确定为破骨细胞.结果:脂联素呈剂量依赖性抑制人成骨细胞护骨素mRNA与蛋白质的表达(P<0.05).脂联素呈剂量依赖性促进人成骨细胞核转录因子κB受体活化素配体mRNA与蛋白的表达(P<0.05).脂联素干预成骨细胞与外周血单核细胞共同培养系统可诱导破骨细胞生成.结论:脂联素可通过诱导成骨细胞核转录因子κB受体活化素配体表达、抑制护骨素表达,诱导破骨细胞生成.
배경:재골조직중,호골소적주요작용시억제파골세포적형성화활성,핵전록인자κB수체활화소배체적주요작용위자격파골세포적분화화활성,억제파골세포적조망.호골소/핵전록인자κB수체활화소배체재파골공능조공중기중요작용.근래연구발현지련소촉진성골세포증식화분화,지련소여골대사상호우련.단지련소대파골세포적작용불명.목적:관찰지련소대성골세포핵전록인자κB수체활화소배체화호골소표체적영향,진일보분석기대파골세포생성적작용.설계、시간급지점:대비관찰실험,우2007-0612008-12재중남대학상아이의원대사내분비연구소실험실완성.재료:외과수술취정상성인가전상극송질골용우세포배양,림상표본유상아의원제공.방법:분별용0,3,10화30 mg/L지련소간예인성골세포48 h,검측호골소화핵전록인자κB수체활화소배체mRNA여단백표체.병용지련소간예성골세포여외주혈단핵세포공동배양계통,관찰기대파골세포생성적영향.주요관찰지표:분별채용형광정량취합매련반응화매련면역흡부법검측인성골세포호골소화핵전록인자κB수체활화소배체mRNA여단백표체.항주석산린산매염색학정위파골세포.결과:지련소정제량의뢰성억제인성골세포호골소mRNA여단백질적표체(P<0.05).지련소정제량의뢰성촉진인성골세포핵전록인자κB수체활화소배체mRNA여단백적표체(P<0.05).지련소간예성골세포여외주혈단핵세포공동배양계통가유도파골세포생성.결론:지련소가통과유도성골세포핵전록인자κB수체활화소배체표체、억제호골소표체,유도파골세포생성.
BACKGROUND: The main role that the osteoprotegedn (OPG) plays in bone tissues is to inhibilate the formation and the activity of osteoclast, while as for the receptor activator of nuclear factor-κB ligand (RANKL), it is to stimulate the differentiation and the activity of osteoclast. Both OPG and RANKL are important for the regulation of osteoclastic function. Recent studies have found the stimulative effects of adiponectin on the proliferation and the differentiation of osteoblast, as well as the coupling between adiponectin and bone metabolism. But effects of adiponectin on osteoclast remain unclear. OBJECTIVE: To investigate the effect of adiponectin on the expression of OPG and RANKL in osteoblast, and to further analyze its effects on the formation of osteoclast. DESIGN, TIME AND SETTING: A contrast observational experiment was conducted in the laboratory of Institute of 2008.MATERIALS: Cancellous bone in anterior superior lilac spine was obtained from adult normal by surgery for cell incubation. Clinical samples were provided by the Xiangya Hospital. METHODS: Human osteoblast was incubated with different doses of adiponcetin (0, 3, 10 and 30 mg/L) for 48 hours, after which OPG and RANKL mRNA and protein expression level was determined. In addition, adiponcetin was added into the co-culture system of osteoblast and peripheral blood monouclear cells for examing its effects on osteoclast formation. MAIN OUTCOME MEASURES: OPG and RANKL mRNA and protein expression in human osteoblast was determined using fluorescent quantitative polymerase chain reaction (PCR) and enzyme linked immunosorbent assay (ELISA). Osteoclast was detected by antitartaric acid acid phosphatase staining. RESULTS: Adiponcetin inhibited osteoblast OPG mRNA and protein expression in a dose-dependent way (P < 0.05). Adiponcetin promoted osteoblast RANKL mRNA and protein expression in a dose-dependent way (P < 0.05). Adiponectin induced the osteoclasts formation in the co-culture system of osteoblast and peripheral blood monouclear cells. CONCLUSION: Adiponectin has the effect of inducing osteoclast formation via stimulating RANKL expression and inhibiting OPG