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目的研究中药舒肝健骨方对人成骨肉瘤MG-63细胞骨形态发生蛋白-2(BMP-2)和骨特异性转录因子(Runx2)基因表达和蛋白合成的影响,探讨其对骨合成代谢的影响机制。方法 MG-63细胞培养及传代,分组药物干预:对照组,阿那曲唑组,舒肝健骨方组,联合用药组,培养48 h后,抽提细胞总RNA,实时荧光定量PCR检测BMP-2和Runx2 mRNA表达;提取细胞总蛋白,蛋白免疫印迹法检测蛋白合成的变化。结果联合用药组和舒肝健骨方组BMP-2和Runx2 mRNA表达上调,2组BMP-2 mRNA相对表达量分别是对照组的1.65、2.34倍,Runx2 mRNA的相对表达量分别是对照组的1.41、2.33倍,差异有统计学意义,阿那曲唑组BMP-2和Runx2 mRNA的表达下调。联合用药组、舒肝健骨方组BMP-2和Runx2蛋白的表达上升,BMP-2蛋白的相对表达量分别是0.75、1.01,Runx2蛋白的相对表达量分别是0.07、0.12,而阿那曲唑组二者的表达降低,与对照组比较虽无统计学意义,但与其余2组比较有统计学意义。结论中药舒肝健骨方对骨合成代谢有促进作用,这可能是通过上调BMP-2及其下游特异性转录因子Runx2 mRNA的表达及蛋白合成而实现的。
Objective To investigate the effect of Shugan Jiangu Fang on gene expression and protein synthesis of bone morphogenetic protein-2 (BMP-2) and bone-specific transcription factor (Runx2) in human osteosarcoma MG- Mechanism of metabolism. Methods MG-63 cells were cultured and passaged, grouping drug intervention: control group, anastrozole group, Shugan Jiangu Fang group and combination group. After cultured for 48 h, the total RNA was extracted and detected by real-time fluorescence quantitative PCR. 2 and Runx2 mRNA expression. Total cellular protein was extracted and the protein synthesis was detected by Western blotting. Results The expression of BMP-2 and Runx2 mRNA in the combination group and Shugan Jiangu Fang group were up-regulated. The relative expression of BMP-2 mRNA in the two groups were 1.65 and 2.34 times of that in the control group respectively. The relative expressions of Runx2 mRNA in control group 1.41 and 2.33-fold respectively. The difference was statistically significant. The expression of BMP-2 and Runx2 mRNA in anastrozole group was down-regulated. The expression of BMP-2 and Runx2 proteins in combination group and Shugan Jiangu Fang group were increased, the relative expression levels of BMP-2 protein were 0.75,1.01, and the relative expression levels of Runx2 protein were 0.07, 0.12, respectively. Anastrozole The expression of the two groups decreased, compared with the control group, although not statistically significant, but compared with the remaining two groups were statistically significant. Conclusion Shugan Jiangu Recipe can promote bone anabolism, which may be through up-regulating BMP-2 and its downstream transcription factor Runx2 mRNA expression and protein synthesis.