1,25二羟维生素D_3对小鼠成骨细胞增殖分化及维生素D受体表达的影响  

Effects of 1,25-Dihydroxyvitamin D_3 on Cell Proliferation,Differentiation and Expression of Vitamin D Receptor in Mouse Osteoblast

作　　者：顾海燕[1] 李婵娟[2] 王全[1] 吴越[3] 郭锡熔[2] 赵德育[1] 

机构地区：[1]南京医科大学附属南京儿童医院呼吸科,南京210008 [2]南京医科大学附属南京妇幼保健医院儿科,南京210004 [3]南京医科大学附属南京儿童医院新生儿科,南京210008

出　　处：《实用儿科临床杂志》2009年第19期1484-1486,1498,共4页Journal of Applied Clinical Pediatrics

基　　金：南京市卫生局医学科技发展项目资助(ZKM06045);南京医科大学重点发展项目资助(06NMU2024)

摘　　要：目的研究不同浓度1,25二羟维生素D3[1,25(OH)2D3]对小鼠成骨细胞增殖分化及维生素D受体(VDR)表达的影响,探讨VDR在成骨细胞增殖分化中的作用。方法在成骨细胞株MC3T3E1培养液中加入不同浓度的1,25(OH)2D3(10-8、10-9、10-10mol/L),培养48h后应用噻唑蓝(MTT)比色法检测其对成骨细胞增殖的影响,采用细胞培养上清中骨钙素(OC)水平分析方法检测1,25(OH)2D3对成骨细胞分化的影响;采用SYBR Green荧光定量PCR方法检测细胞VDR基因mRNA表达,Western blot法检测细胞VDR蛋白水平表达。结果1.小鼠成骨细胞在1,25(OH)2D3处理48h后,各实验组(10-8、10-9、10-10mol/L组)吸光度均低于对照组,差异均有统计学意义(Pa<0.05);2.1,25(OH)2D3处理后,10-8、10-9mol/L组细胞上清中OC水平均高于对照组(Pa<0.05),而10-10mol/L组OC水平及VDR基因mRNA表达量与对照组比较差异均无统计学意义(Pa>0.05);3.荧光定量PCR及Westernblot结果显示10-8、10-9mol/L处理组细胞VDR基因mRNA及蛋白表达均显著高于对照组(Pa<0.05),10-10mol/L组与对照组比较无显著性差异(P>0.05)。结论1,25(OH)2D3可显著抑制成骨细胞增殖、促进成骨细胞分化,且可促进成骨细胞VDR表达,提示VDR信号通路在成骨细胞的增殖分化中可能起一定作用。Objective To study the effect of different concentration of 1,25 - dihydroxyvitamin D3 [ 1,25 （ OH ） 2 D3 ] on cell proliferation,differentiation and the expression of vitamin D receptor （VDR） in mouse MC3T3E1 osteoblast. Methods Osteoblast were cultured in medium with different concentrations of 1,25 （OH） 2 D3. Incubated for 48 h, cell proliferation of osteoblast were examined by MTT reduction assay （mono -nuclear cell diree cytotoxicity assay）, the osteocalcin （OC） levels in cell medium were detected by ELISA, and the expression of VDR mRNA and protein were examined by using SYBR Green real - time PCR and Western blot, respectively. Results 1. After incubation with 1,25 （ OH ） 2 D3 for 48 h, the number of MC3 T3 E 1 osteoblast was significantly less than that in control group （ P 〈 0.05 ）. 2. OC concentra- tions in culture medium of the 10 ^-8, 10 ^-9 mol/L experimental groups were higher than those in control group （ Pa 〈 0.05 ） ; while the 10^ -10mol/L group had no significant difference compared with control group （ P 〉 0.05 ）. 3. SYBR Green real - time PCR and Western blot results showed that the expression of VDR mRNA as well as VDR protein of osteoblast in 10^-8,10^-9 mol/L experimental groups were significantly higher than those in control group （P 〈 0.05 ）. However, there was no significant difference observed between 10^- 10 mol/L group and control group （ P 〉 0.05 ）. Conclusions Cell proliferation of mouse osteoblast can be inhibited, while the cell differentiation was promoted by 1,25 （OH）2 D3.1,25 （OH）2D3 up- regulated the expression of VDR in mouse osteoblast, which suggested that the VDR signal pathway may play some role in proliferation and differentiation of osteoblast.

关 键 词：1 25二羟维生素D3 成骨细胞 细胞分化 维生素D受体 

分 类 号：R723[医药卫生—儿科] R726.8[医药卫生—临床医学]