CAJ | 학술논문

背景:骨质疏松涉及破骨细胞分化成熟和成骨类细胞凋亡两方面,肿瘤坏死因子α能诱导多种细胞凋亡且效率差异很大,骨保护素能抑制破骨细胞成熟,但对成骨类细胞凋亡的作用未见报道.目的:探讨肿瘤坏死因子α能否诱导成骨类细胞凋亡及骨保护素能否抑制其凋亡.设计:以成骨细胞和MG63细胞作为实验对象、以鼠WEHI 164细胞系用于阳性对照的观察对比分析.单位:解放军总医院全军口腔医学研究所、美国匹茨堡医学院退伍军人医疗中心.材料:本实验于2001-01/2003-12在Dr.Blair实验室和口腔研究所实验室完成.选择商品化细胞系,RPMI 1640培养液和各种相关蛋白用于实验.方法:用人间充质干细胞在分化培养液中培养21 d使之成为成骨细胞,与MG63一起用于凋亡实验.Annexin V和原位末端标记法检测肿瘤坏死因子α诱导成骨类细胞凋亡.主要观察指标:全部细胞数及凋亡细胞数.结果:和FasL诱导细胞凋亡一样,肿瘤坏死因子α能引起MG63骨肉瘤细胞,间充质干细胞和成骨细胞的凋亡,并表现为明显的浓度依赖和时间依赖.低浓度肿瘤坏死因子α(170～500 pmol/L)作用于细胞2～4 h就显示了较明显的凋亡,而骨保护素在0.45～1.5 nmol/L浓度时,几乎完全抑制了500 pmol/L肿瘤坏死因子α诱导的凋亡.成骨细胞分泌骨保护素,而破骨细胞及破骨前体细胞产生肿瘤坏死因子α,它们相互作用降低成骨细胞的凋亡.结论:肿瘤坏死因子α能诱导成骨类细胞凋亡,骨保护素能够抑制肿瘤坏死因子α的凋亡诱导作用,从抑制破骨细胞成熟和成骨细胞凋亡两方面表现骨保护素防治骨质疏松的作用机制.
배경:골질소송섭급파골세포분화성숙화성골류세포조망량방면,종류배사인자α능유도다충세포조망차효솔차이흔대,골보호소능억제파골세포성숙,단대성골류세포조망적작용미견보도.목적:탐토종류배사인자α능부유도성골류세포조망급골보호소능부억제기조망.설계:이성골세포화MG63세포작위실험대상、이서WEHI 164세포계용우양성대조적관찰대비분석.단위:해방군총의원전군구강의학연구소、미국필자보의학원퇴오군인의료중심.재료:본실험우2001-01/2003-12재Dr.Blair실험실화구강연구소실험실완성.선택상품화세포계,RPMI 1640배양액화각충상관단백용우실험.방법:용인간충질간세포재분화배양액중배양21 d사지성위성골세포,여MG63일기용우조망실험.Annexin V화원위말단표기법검측종류배사인자α유도성골류세포조망.주요관찰지표:전부세포수급조망세포수.결과:화FasL유도세포조망일양,종류배사인자α능인기MG63골육류세포,간충질간세포화성골세포적조망,병표현위명현적농도의뢰화시간의뢰.저농도종류배사인자α(170～500 pmol/L)작용우세포2～4 h취현시료교명현적조망,이골보호소재0.45～1.5 nmol/L농도시,궤호완전억제료500 pmol/L종류배사인자α유도적조망.성골세포분비골보호소,이파골세포급파골전체세포산생종류배사인자α,타문상호작용강저성골세포적조망.결론:종류배사인자α능유도성골류세포조망,골보호소능구억제종류배사인자α적조망유도작용,종억제파골세포성숙화성골세포조망량방면표현골보호소방치골질소송적작용궤제.
BACKGROUND: Osteoporosis involves two aspects, osteoblast apoptosis and osteoclast differentiation and maturation. The apoptotic effect of tumor necrosis factor-α varies in different cell lines and it remains unclear whether osteoprotegerin has inhibitory effects on the apoptosis of osteoblastic cells.OBJECTIVE: To study the interaction between tumor necrosis factor-α and osteoprotegerin on the apoptosis of osteoblastic cells.DESIGN: Observational comparative study based osteoblasts and MG63 cells as subjects and murine WEHI 164 cells as positive controls.SETTING: Institute of Oral Medicine, General Hospital of Chinese PLA, and Veterans' Affairs Medical Center, Pittsburgh.MATERIALS: The experiment was conducted in the Dr. Blair' s Laboratory and Oral Institute from January 2001 to December 2003. Commercial cell group, RPMI 1 640 culture medium and various related proteins were selected in this study.METHODS: Osteoblasts were differentiated from human mesenchymal stem cells in the culture medium. and were ready for the study on apoptosis with MG63. Annexin V and TUNEL analysis were used to assay osteoblasts'apoptosis induced by tumor necrosis factor-α.MAIN OUTCOME MEASURES: The number of total cells and apoptotic cells.RESULTS: As FasL induced apoptosis, tumor necrosis factor-α could obviously induce the apoptosis of MG63 osteogenic cells, mesenchymal stem cells and osteoblasts in concentration-and time-dependent manner. Cells showed obvious apoptosis after 2 -4 hours' effect by tumor necrosis factor-α in low concentration ( 170 - 500 pmol/L); osteoprotegerin, on the other hand, at 0. 45 - 1.5 nmol/L inhibited the apoptosis induced by tumor necrosis factor-α of 500 pmol/L. Osteoblasts could secrete osteoprotegerin, whereas osteoclasts and precursor of osteoclasts could produce tumor necrosis factor-α and work together to reduce the apoptosis of osteoblasts.CONCLUSION: Tumor necrosis factor-α can effectively induce the apoptosis of osteoblastic cells while osteoprotegerin has the ability to inhibit it. The mechanism of osteoprotegerin on osteoporosis involves at least two aspects, by inhibiting the maturation of osteoclasts and the apoptosis of osteoblasts.