淡豆豉异黄酮对大鼠成骨细胞增殖及雌激素受体表达的影响

Effects of isoflavone from Sojae Semen Preparatum on the proliferation and expression of estrogen receptor in rat osteoblasts

目的观察淡豆豉异黄酮对大鼠成骨细胞增殖及雌激素受体表达的影响。方法采用改良组织块法分离培养新生SD大鼠颅骨成骨细胞,分别给予不同质量浓度淡豆豉异黄酮（1、10、100、1 000μg/L）、1 000μg/L淡豆豉异黄酮＋10 nmol/L雌激素受体拮抗剂ICI 182780、1 000μg/L大豆异黄酮进行干预,以1 nmol/L 17β-雌二醇作为阳性对照,MTT法检测成骨细胞增殖情况;RT-PCR和Western blot法分别检测成骨细胞ERβmRNA及蛋白表达水平。结果MTT结果显示,淡豆豉异黄酮在1~1 000μg/L范围内促进成骨细胞增殖;RT-PCR和Western blot结果显示,1~1 000μg/L淡豆豉异黄酮上调ERβmRNA及蛋白表达水平。而1 000μg/L淡豆豉异黄酮的上述作用能够被雌激素受体拮抗剂所拮抗。结论淡豆豉异黄酮能够促进大鼠成骨细胞的增殖,此作用可能是通过雌激素受体ERβ途径实现的。

AIM To study the effects of isoflavone from Sojae Semen Preparatum（ ISSP） on the proliferation and expression of estrogen receptor（ ER） in rat osteoblasts. METHODS Modified tissue block method was used to separate and culture calvarial osteoblasts from newborn SD rats. 1,10,100,1 000 μg / L ISSP,1 000 μg / L ISSP ＋ 10 nmol / L ER antagonist ICI 182780 and 1 000 μg / L soy isoflavone were added to the culture system,respectively. 17β-estradiol at the concentration of 1 nmol / L was used as a positive control. The proliferation of osteoblasts was detected by MTT method,the expressions of ERβ mRNA and protein were detected by RT-PCR and Western blot,respectively. RESULTS The results of MTT examination revealed that the proliferation of osteoblasts was promoted in the ISSP concentration ranges of 1- 1 000 μg / L. The results of RT-PCR and Western blot examination revealed that ISSP（ 1- 1 000 μg / L） could up-regulate ERβ mRNA and protein expressions. The above functions of ISSP at the concentration of 1 000 μg / L could be inhibited by adding ER antagonist. CONCLUSION ISSP can promote the proliferation of rat osteoblastes in vitro,and this promotion effect may be mainly related to ERβ pathway.