川芎嗪对骨髓间充质干细胞表达心肌发育早期相关基因及基质源性细胞因子受体的影响

Effects of Tetramethylpyrazine on Expression of Early Myocardiac Development Related Genes and Stromal Cell-derived Factor Receptors in Marrow Mesenchymal Stem Cells

目的观察川芎嗪对骨髓间充质干细胞(marrow mesenchymal stem cells,MMSCs)表达心肌发育早期基因GATA结合蛋白4(GATA binding 4,GATA-4)、NK2相关转录因子5位点(NK2transcription fac-tor related,locus 5,Nkx2-5)和肌球蛋白重链(myosin heavy chain,MHC)mRNA的影响,以及对基质源性细胞因子受体趋化因子(C-X-C基序)受体4(chemokine(C-X-C motif)receptor 4,CXCR-4)mRNA表达的影响。方法用含不同浓度的川芎嗪培养液培养MMSCs,培养1、7、21d后,用RT-PCR法检测MMSCs各基因mRNA的表达水平。结果①MMSCs组以及经不同浓度川芎嗪诱导后均未表达GATA-4、Nkx2-5和MHC mRNA;心肌阳性对照组表达相关基因。②MMSCs组和5-氮胞苷(5-azacytidine,5-AZA)组在早期CXCR-4的表达水平略有升高,7d后逐渐下降。不同浓度川芎嗪组MMSCs表达CXCR-4mRNA的水平随着干预时间的延长逐渐增加。结论①川芎嗪不能促进MMSCs表达心肌发育早期相关基因;②川芎嗪可增加MMSCs表达CXCR-4mRNA,且呈剂量依赖性。

objective To observe the effects of tetramethylpyrazine （TMP） on the expression of early myo- cardiac development related genes in marrow mesenchymal stem cells （MMSCs）, including GATA bindingprotein 4 （GATA-4）, NK2 transcription factor related, locus 5 （Nkx2-5）, and myosin heavy chain （MHC）, as well as chemokine （C-X-C motif） receptor 4 （CXCR-4） , a receptor of stromal cell derived fac tor-1. Methods MMSCs were cultured in medium containing different dose TMPs in vitro. After 1-, 7-, and 21-day culturing, the expression levels of all above-mentioned genes mRNA in MMSCs were detected using reverse transcription PCR. Results （1）There were no expression of GATA 4, Nkx2-5, and MHC mRNA in MMSCs group and different dose TMPs treated groups, but there were expressions of above- mentioned genes mRNA in the positive control, or cardiac muscle group. （2）The expression levels of CX- CR-4 mRNA were slightly increased in the early stage and gradually decreased 7 clays later in the MMSCs group and the 5-azacytidine treated group. The expression levels of CXCR-4 mRNA increased in a time de- pendent manner in different dose TMPs treated groups. Conclusion （1） TMP cannot upregulate the ex- pression of early myocardiac development related genes in MMSCs; （2）TMP can upregulate the expression of CXCR-4 mRNA in MMSCs in a dose-dependent manner.