孕激素对成骨细胞增殖和分化的影响

Effects of progestin on proliferation and differentiation of osteoblasts

目的 观察孕酮和左炔诺孕酮对正常妇女成骨细胞和成骨肉瘤MG 6 3细胞增殖和分化的影响。方法 采用四甲基偶氮唑蓝法 ,检测应用不同浓度孕酮及左炔诺孕酮刺激上述两种成骨细胞增殖的情况 ;并采用半定量RT PCR及免疫组化法 ,测定c fos、c jun基因及其蛋白和骨钙素mRNA表达。结果 以浓度为 0mol/L的孕酮和左炔诺孕酮为对照 ,10 -10 、10 -8、10 -6mol/L孕酮分别增加人成骨细胞及MG 6 3细胞增殖达 9 8%、2 3 0 %、32 8%及 7 8%、15 6 %、2 1 8% ;10 -10 、10 -8、10 -6mol/L左炔诺孕酮分别增加人成骨细胞及MG 6 3细胞增殖达 12 5 %、2 1 9%、32 8%及9 6 %、14 4 %、2 3 0 % ;孕酮及左炔诺孕酮促进两种成骨细胞c fos、c jun基因及其蛋白的表达 ,表达呈剂量依赖性 ;10 -10 、10 -8、10 -6mol/L孕酮和左炔诺孕酮增加人成骨细胞骨钙素mRNA表达分别为 12 2 %、2 3 7%、4 5 5 %及 18 6 %、32 1%、6 0 3% ;孕酮和左炔诺孕酮对MG 6 3细胞骨钙素mRNA的表达无影响。结论 孕酮和左炔诺孕酮可刺激成骨细胞增殖 ,增加c fos、c jun基因及其蛋白的表达 。

Objective To study the effects of progesterone and levonorgestrel (LNG) on proliferation and differentiation of human normal osteoblasts and human osteosarcoma cell line MG 63 Methods Cell proliferation was tested by 3 [4,5 Dimethylthiazolzyl] 2,5 Diphenyl Tetrazolium Bromide (MTT) assay The expression of c fos, c jun and osteocalcin was measured by semi quantitative reverse transcription polymerase chain reaction (RT PCR) or immunocytochemistry Results Compared to the control group, progesterone at concentrations of 10 -10 mol/L, 10 -8 mol/L and 10 -6 mol/L increased the proliferation of human osteoblasts by 9 8%, 23 0% and 32 8% respectively and that of MG 63 cells by 7 8%, 15 6% and 21 8% respectively LNG at concentrations of 10 -10 mol/L, 10 -8 mol/L and 10 -6 mol/L increased the proliferation of human osteoblasts by 12 5%, 21 9% and 32 8% respectively and that of MG 63 cells by 9 6%, 14 4% and 23 0% respectively Progesterone and LNG upregulated c fos and c jun mRNA and protein in a dose dependent manner Likewise,progesterone and LNG increased the expression of osteocalcin mRNA in human osteoblasts by 12 2%,23 7%,45 5% and 18 6%,32 1%,60 3% respectively The effects of LNG were greater than those of progesterone Meanwhile,the expression of osteocalcin mRNA in MG 63 cells was not influenced by either progesterone or LNG Conclusion Progesterone and LNG stimulate proliferation of osteoblasts and increase the expression of c fos, c jun and osteocalcin