目的 探讨资冲颗粒(熟地、菟丝子等)诱导大鼠间充质干细胞向生殖细胞分化相关基因表达的影响.方法 制备大鼠含资冲颗粒血清及大鼠空白血清对大鼠间充质干细胞进行干预.显微镜观察细胞形态变化情况;实时荧光定量PCR测定干预后细胞内生殖分化相关Oct4、Stra8、Itga6、Itgb1、Scp3、Gdf9、Zp1、Zp3基因的表达;MTT法检测细胞增殖情况;运用统计软件分析两组数据.结果 镜下观察72 h干预未发现含药血清和空白血清对细胞形态有明显影响;实时荧光定量PCR测定表明含药血清对Stra8、Scp3、Gdf9基因表达有显著影响(P<0.05),对其它基因表达的影响则不显著(P>0.05);MTT法显示5d干预两组细胞增殖无差别(P>0.05).结论 资冲颗粒大鼠含药血清具有诱导大鼠间充质干细胞向生殖细胞方向分化的潜力.
AIM To investigate the effect of Zichong Granules (Rehmanniae Radix praeparata, Cuscutae Se- men, etc) to induce rat mesenchymal stem cells to germ cells. METHODS The rat serum containing Zichong Granules and the blank rat serum were used to intervene rat mesenchymal stem cells. The change of the cellular morphology was observed by microscopy. The expressions of eight marker genes ( Oct4, Stra8, Itga6, Itgbl, Scp3, Gdjg, Zpl, Zp3 ) related to reproductive differentiation were examined by real-time fluorescence quantita- tive PCR. The change of the cellular proliferation was also determined by MTr assay. RESULTS The microsco- py showed that there were few differences of cellular morphology between Zichong Granules containing serum and the blank rat serum in 72 hours of the intervention. However, results of real-time fluorescence quantitative PCR in- dicated that the intervention significantly influenced the transcriptions of Stra8, Scp3, GdJ9 ( P 〈 O. 05 ), but there was no significant influence on the transcriptions of Oct4, Itga6, Zpl and Zp3. Moreover, the MTr assay showed that the rat medicated serum had little effect on the cellular proliferation during the five-days intervention (P 〉 0. 05 ). CONCLUSIONS Zichong Granules have the potential to induce rat mesenchymal stem cells to differenti- ate to germ cells.