补肾方对成骨细胞生长因子TGF-β_1 mRNA表达的影响

THE INFLUENCE OF KIDNEY-TONYFYING METHOD ON mRNA EXPRESSION OF GROWTH FACTOR TGF-β_1 IN RAT OSTEOBLASTS

为探讨补肾方对成骨细胞转化生长因子 - β1 (TGF- β1 ) m RNA表达的影响 ,将体外分离培养的新生 SD大鼠的颅骨成骨细胞加入不同浓度的补肾中药密骨片药液 (10 0～ 10 0 0 0μg· ml- 1 )及阳性对照药物重组碱性成纤维细胞生长因子 (rb FGF)。2 4小时后 ,利用自行制备的地高辛标记的鼠 TGF- β1 c DNA探针进行成骨细胞的核酸分子原位杂交分析 ,以其细胞内杂交阳性颗粒的平均光密度值代表 TGF-β1 m RNA的表达水平。结果显示随着密骨片药液浓度的增加 ,成骨细胞内 TGF-β1 m RNA表达明显高于阴性对照组 ;但 5 0 0 0和 10 0 0 μg·m l- 1的密骨片药液组的 TGF- β1 光密度值与 5 ng· ml- 1的 b FGF组比较无明显差异 (P>0 .0 5 )。表明补肾中药密骨片可刺激成骨细胞 TGF-β1 的分泌和合成 ,从而促进骨形成和抑制骨吸收。这可能是该方能有效地防治骨质疏松的疗效机理之一。

To investigate the effect of kidney tonyfying method on mRNA expression of growth factor TGF β 1 in rat osteoblasts, the calvarial osteoblasts of new born SD rats were cultured isolated in 6 culture bottles, then different concentration(100 ,1000,5000,10000μg/ml) solution of kidney tonyfying Traditional Chinese Medicine Migu Tablet and recombinant basic fibroblast growth factor (rbFGF, 5μg/ml) were added to the five bottles respectively (served as positive control group), and the other bottle was made negative control group. Twentyfour hours later the osteoblasts were made situnucleic acid hybridixation by Digoxigenin labelled TGF β 1 cDNA probes, and the levels of the mRNA expression of TGF β 1 were represented by the mean optical density values of stained granules hybridixated positively. The results showed with the increase of the concentration of Migu Tablet solution, the levels of the mRNA expression of TGF β 1 of Migu Tablet were evidently more and more high than those of negative control group, while the mean optical density values of TGF β 1 in 5000μg/ml Migu Tablet solution and those in positive control group made no difference. So Migu Tablet could promote bone formation and inhibit bone resorption by stimulating the secretion and synthesis of TGF β 1 in osteoblasts, which was the mechanism of Migu Tablet in preventing and treating osteoporosis.