Ca~(2+) signaling mediated salidrosides promotes directional differentiation of mouse bone marrow mesenchymal stem cells into nerve cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Tranditional Chinese medicine,which possesses anti-oxidation properties,can promote directional differentiation of mouse bone marrow mesenchymal stem cells(BMMSCs)into nerve cells.Salidrosides,as the effective constituent of Rhodiola,have strong anti-oxidation function.OBJECTIVE:To investigate the molecule mechanism of salidrosides induced differentiation of mouse BMMSCs into nerve cells.METHODS:When in vitro cultured BMMSCs reached 80% confluency,the cells were assigned into 3 groups.Cells in the control group were cultured by complete culture medium;those in the induction and positive control groups were cultured by complete culture medium adding 20 mg/L salidrosides or 0.1 mg/L nerve growth factors(NGF).The related gene and protein of nerve cells were detected using RT-PCR and Western blot method at 12 hours after culture.After that,the cells in the induction group were divided into 3 groups,the blocking agents EGTA(Ca~(2+) chelator),Nifedipine(L-type Ca~(2+) channel blocker)and LY294002(IP3 receptor blocking pharmacon)were applied to block the cellular Ca2* signal pathway respectively for 12 hours.RT-PCR and Western blot methods were used to study the signal transduction of the salidrosides.RESULTS AND CONCLUSION:①The expression of neuron specific enolase(NSE),B-Tubulin III,Nurri mRNA could be found ir the induction and positive control groups,instead of the control group;The expression abundance of the positive control group was smaller than that of the induction group.The expression abundance of GFAP mRNA was very low in each group,but the c-fos mRNA was expressed abundantly in the induction group.②Compared with the positive control group,the induction group could promote the NSE expression obviously,which was no expressed in the blank control group.?The expression of NSE and Nurri were conspicuously down-regulated when the Ecto Ca~(2+) and L-type Ca~(2+) channel and IP3 receptor were blocked respectively.Salidrosides can induce the differentiation BMMSCs into nerve cells. Ca~(2+) signaling and IP3 dependent Ca~(2+) signaling pathway play an important role in transduction salidrosides signal in BMMSCs differentiation.