肉桂酸对自发转化后大鼠骨髓间充质干细胞增殖和分化的影响

Effects of Cinnamic Acid on the Proliferation and Differentiation of Rat Spontaneous Transformed Mesenchymal Stem Cells

目的:研究肉桂酸(cinnamic acid,CINN)对自发转化后大鼠骨髓间充质干细胞(transformed mesenchymal stem cells,TMCs)增殖、分化、细胞周期及原癌基因c-fos表达的影响。方法:不同浓度(1、3、5 mmol/L)CINN处理TMCs,四甲基偶氮唑蓝(MTT)法测定细胞生长抑制率,流式细胞仪分析细胞周期,ALP钙钴染色及western blot方法观察TMCs向成骨细胞的分化能力,端粒酶活性TRAP实时定量检测试剂盒检测细胞端粒酶活性,RT-PCR及Western blot方法检测原癌基因c-fos的表达。结果:不同浓度的CINN作用后TMCs生长均明显受到抑制,5 mmol/L CINN处理后第5天抑制率最高达(63.41±3.69)%。CINN作用后钙钴染色细胞阳性率明显高于对照组(P<0.01),且Runx2表达量显著升高。CINN处理组细胞端粒酶活性降低,为对照组的43%。RT-PCR及Western-blot显示CINN作用后c-fos基因的mRNA及蛋白表达水平均明显下降。结论:肉桂酸可抑制大鼠TMCs增殖,并促进其向成骨细胞分化,其机制可能与降低TMCs端粒酶活性及c-fos基因的表达有关。

：Objective To investigate the effects of cinnamic acid on the proliferation, differentiation, cell cycle and c-fos gene expression of transformed mesenchymal stem cells （ TMCs）. Methods TMCs were treated with different concentrations of cinnamic acid（ 1,3,5 mmol/L）.Then the growth inhibitory ratio was detected by MTT method,the cell cycle was analyzed with flow cytometer,the differentiation potency of TMCs was observation with ALP staining and Western-blot, the telomerase activity was detected by TRAP real-time quantitative kits, the expression of c-fos gene was examined with RT-PCR assay and Western-blot.Results The growth of TMCs was obviously inhibited after treating with different concentrations of cin- namic acid, the inhibitory rate was high to （63.41±3.69）% at 5th day after treatment.The positive rate of ALP staining in cinnamic acid treating TMCs was obviously higher than that of control（ P〈0.01 ） , and the expression of Runx2 was signifi- cantly increased.The telomerase activity of treated TMCs was decreased to 43% of that in control group.The results of RT- PCR and western-blot demonstrated that the mRNA expression of c-fos gene in TMCs decreased after treating with cinnamic acid.Conclusion Cinnamic acid could inhibit the proliferation of TMCs and induce its differentiation to osteoblasts,which may be related to decrease the telomerase activity and c-fos gene expression of TMCs.