1α,25(OH)_2D_3对大鼠成骨细胞КB受体活化因子配体及骨保护素蛋白表达的影响

EFFECTS OF 1α,25(OH)_2D_3 ON THE EXPRESSION OF RECEPTOR ACTIVATOR OF NF-КB LIGAND AND OSTEOPROTEGERIN IN RAT OSTEOBLASTS IN VITRO

目的研究1α,25-二羟维生素D3(1α,25(OH)2D3)对体外培养3～4d的SD大鼠成骨细胞(OB)核因子κB受体活化因子配体(RANKL)及骨保护素(OPG)蛋白和mRNA表达的影响。方法1α,25(OH)2D3作用24、48、72h,分别采用ELISA及FQ-PCR法测定。结果10、100nmol/L1α,25(OH)2D3较对照及1nmol/L显著或极显著促进RANKL蛋白及mRNA表达;1、10nmol/L1α,25(OH)2D3较对照显著或极显著促进OPG蛋白及mRNA表达,而100nmol/L则极显著抑制OPGmRNA表达;RANKLmRNA/OPGmRNA及RANKL/OPG显示,1nmol/L组与对照组差异不显著,10、100nmol/L组RANKLmRNA/OPGmRNA及RANKL/OPG极显著高于对照组和1nmol/L组。结论低浓度1α,25(OH)2D3(1nmol/L)对骨更新作用不明显,而中、高浓度1α,25(OH)2D(310、100nmol/L)能通过上调RANKLmRNA/OPGmRNA及RANKL/OPG比例,促进OC的生成及骨吸收功能,增强骨更新。

Objective To investigate the effect of 1α,25-dihy-droxyvitamin D3 （1α,25（OH）2D3） on the expression of receptor activator of NF-КB ligand （RANKL）,osteoprotegerin （OPG） and RANKL mRNA,OPG mRNA,of rat osteobalsts （OB）. Method The expression of RANKL and OPG was detected by the method of Immunohistochemistry and ELISA. RANKL mRNA and OPG mRNA were determined through FQ-PCR. Results Compared with the control group and the group with 1 nmol/L 1α,25（OH）2D3,10 and 100 nmol/L 1α,25（OH）2D3 can significantly induce the expression of RANKL and RANKL mRNA. 1,10 nmol/L 1α,25（OH）2D3 can stimulate the expression of OPG and OPG mRNA significantly,while 100 nmol/L 1α,25（OH）2D3 can inhibit the expression of OPG mRNA significantly. There were no difference in the rate of RANKL/OPG and RANKL mRNA/OPG mRNA between the control group and the group with 1 nmol/L 1α,25（OH）2D3,however the rate of RANKL/OPG and RANKL mRNA/OPG mRNA in the group with 10,100 nmol/L were higher than the control group and the group with 1 nmol/L 1α,25（OH）2D3. Conclusion Lower dosage of 1α,25（OH）2D3 （1 nmol/L） had no significant effect on bone turnover,but higher dosage of 1α,25（OH）2D3 （10,100 nmol/L） can enhance bone turnover through facilitating the formation and activity of osteoclasts via enhancing RANKL mRNA/OPG mRNA and RANKL/OPG..