1,25(OH)2D3对体外培养大鼠成骨细胞RANKL/OPG mRNA表达的影响-东南大学学报医学版

1,25(OH)2D3对体外培养大鼠成骨细胞RANKL/OPG mRNA表达的影响

单位：东南大学附属中大医院,骨科,江苏,南京,210009

分类号：R589.5

出版年·卷·期（页码）：2007·26·第一期（32-35）

摘要：

目的:观察1,25(OH)2D3对体外培养的大鼠成骨细胞RANKL/OPG mRNA表达的影响,探讨其促进骨吸收的作用机制.方法:体外分离培养大鼠成骨细胞,分别观察不同浓度1,25(OH)2D3及用1,25(OH)2D3处理不同时间对RANKL/OPG mRNA表达的影响,RANKL/OPG mRNA表达采用半定量逆转录聚合酶链反应(RT-PCR)方法测定.结果:1,25(OH)2D3呈时间和剂量依赖性地刺激大鼠成骨细胞RANKL mRNA的表达,而抑制OPG mRNA的表达,使RANKL/OPG值增高.结论:1,25(OH)2D3通过调节成骨细胞RANKL/OPG的基因表达,从而发挥促进破骨细胞介导的骨吸收作用.

Objective To observe the effect of 1,25(OH)2D3on the expression of RANKL and osteoprotegerin(OPG) mRNA in primary rat osteoblasts.Methods Primary osteoblastic cells were obtained from the calvaria of newborn SD rats by digestion with 0.1% collagenase and cultured in PRMI 1640 medium supplemented with 15% fetal bovine serum.Cells were treated with 1,25(OH)2D3 according to the experimental design.Total cellular RNA was isolated with Trizol.Semiquantitative reverse transcription RCR method was used to screen the level of RANKL and OPG mRNA.Results RANKL mRNA expression in the calvarial osteoblasts was elevated dose-dependently following 1,25(OH)2D3 treatment and reached its peak level at a concentration of 10-8mol·L-1(P<0.01).1,25(OH)2D3 inhibited OPG mRNA levels by 60% at a concentration 10-7mol·L-1 after 48h(P<0.01).1,25(OH)2D3(10-7mol·L-1)time-dependently resulted in a upregulation of RANKL mRNA levels and a downregulation of OPG mRNA peaking both at 48h(P<0.01).Conclusions 1,25(OH)2D3enhanced RANKL and inhibited OPG mRNA expression of primary rat osteoblasts in a time-and dose-dependent fashion.By altering the relative expression of OPG and RANKL mRNA,1,25(OH)2D3 might stimulate the osteoclastic bone resorption.

参考文献：

[1] JMI E, NAKAMURA I, AMANO H. Osteoclast function is activated by osteoblastic cells through a mechanism involving cell-to-cell contact, 1996
[2] SIMONET S W, LACEY D L, DUNSTAN C R. Osteoprotegerin:a novel secreted protein involved in the regulation of density. 1997. doi:10.1016/S0092-8674(0)80209-3
[3] LACEY D L, TIMMS E, TAN H L. Osteoprotegerin ligand is a cytokine that regulaties osteoclast differentiation and activation. 1998. doi:10.1016/S0092-8674(0)81569-X
[4] HSU H, LACEY D L, DUNSTAN C R. Tumor necrosis factor receptor family member RANK mediates osteoclast differentiation and activation induced by osteoprotegerin ligand. 1999(7). doi:10.1073/pnas.96.7.3540
[5] YASUDA H, SHIMA N, NAKAGAWA N. Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclasteogenesis-inhibitory factor and is identical to TRANCE /RANKL. 1998(7). doi:10.1073/pnas.95.7.3597
[6] 张晔, 吴小涛, 宋萍. 1,25(OH)2D3对体外培养成骨细胞骨保护素mRNA表达的影响. 中华老年医学杂志2002(6)
[7] ECAROT B, GLORIEUX F H, MICHEL R. Osteoblast isolated from mouse calvaria initiate matrix mineralization in culture. 1983. doi:10.1083/jcb.96.3.639
[8] THOMAS G P, BAKER S U K, EISMAN J A. Changing RANKL/OPG mRNA expression in differentiating murine primary osteoblasts. 2001(2). doi:10.1677/joe.0.1700451
[9] OWEN T A, ARONOW M S, BARONE L M. Pleiotropic effects of vitamin D on osteoblast gene expression are related to the proliferative and differentiated state of the bone cell phenotye:dependency upon basal levels of gene expression,duration of exposure,and bone matrix competency in normal rat osteoblast cultures, 1991
[10] SUDA T, UENO Y, FUJII K. Vitamin D and bone. 2003(2). doi:10.1002/jcb.10331
[11] SUDA T, TAKAKGASHI N, UDAGAWA N. Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families. 1999(3). doi:10.1210/er.20.3.345
[12] HOFAUER L C, GORI F, RIGGS B L. Stimulation of osteoprotegerin ligand and inhibition of osteoprotegerin production by glucocorticoids in human osteoblastic lineage cells:potential paracrine mechanisms of glucocorticoid-induced osteoporosis. 1999

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