六味地黄丸、金匮肾气丸和健骨二仙丸对大鼠BMSCs向脂肪细胞分化相关基因表达的影响

The effect of Liuweidihuang Wan,Jinkuishenqi Wan and Jianguerxian Wan differentiation-related genes expression in the process of rat BMSCs adipogenic differentiation

目的:研究补肾益气功效的六味地黄丸、金匮肾气丸和健骨二仙丸对大鼠骨髓间充质干细胞(BMSCs)成脂分化的影响。方法:采用全骨髓贴壁法培养BMSCs,补肾阴、补肾阳和补肾健骨方药(代表方分别为六味地黄丸、金匮肾气丸和健骨二仙丸)含药血清干预BMSCs成脂分化过程,采用反转录-实时荧光定量技术(RT-q real-time PCR),观察成脂分化相关基因Lpl、Fabp4和PparγmRNA表达水平的变化。结果:与10%正常大鼠血清相比较,Fabp4 mRNA在补肾阴组、补肾阳组(10%含药血清)表达水平均下降33%,但无统计学意义;Lpl mRNA在补肾阳组和补肾健骨组(10%含药血清)表达水平下降,其中补肾阳组具有统计学意义。PparγmRNA在其它各组表达无显著性变化。结论:六味地黄丸、金匮肾气丸和健骨二仙丸对转录因子表达水平无显著改变,但对其下游靶基因表达水平有显著下调作用,金匮肾气丸对成脂分化过程中成脂相关基因下调作用最为明显,推测其抑制骨髓间充质干细胞成脂分化能力较其它功效的方药更强。

To study the efficacy of tonifying Qi of the kidney on rat bone marrow mesenchymal stem cells （BMSCs） adipogenic differentiation. Methods ： Whole bone marrow were cultured, to obtain BMSCs by anchorage. Then BMSCs were treated with Liuweidihuang Pill, or Jinguishenqi Pill, or Jianguerxian Pill-containing serum （on behalf of tonifying kidney-YIN, tonifying kidney-YANG and kidney-tonifyng bone-invigorating formula, respectively） in the process of rat BMSCs adipogenic differentiation. Reverse transcription, real time-fluorescent quantitative-PCR （RT real time-FQ-PCR） was used to determine Lpl, Fabp4 and PPAR~/mRNA expression level. Results ： As compared to the control group（ 10% serum）, expression level of Fabp4 mRNA in either the kidney-Yin groups or the kidney-YANG group（ 10% drug- containing serum） decreased by 33%, but no statistical significance; Lpl mRNA expression levels in the kidney-YANG group and kidney- tonifyng bone-invigorating group also decreased, of the total kidney-YANG group was statistically significant （ P 〈 0.05 ）. Change of Ppar3, mRNA expression was not significant in all groups. Conclusions ：Significant change of transcription factor expression level does not occur, but a significant down-regulation of its downstream target genes exists in the process of adipogenic differentiation by Chinese medicine for toni- fying Qi of the kidney. Tonifying Kidney-YANG is most effective in the process, so this indicates that Tonifying kidney-YANG inhibits adipo- genic differentiation of BMSCs more effective than other formulas.