Differentiation And Molecular Mechanism From Mouse Bone Marrow Mesenchymal Stem Cells To Nerve Cells Induced By Salidroside In Vitro

【Objective】According to large number of documents, drugs, which possess anti-oxidation properties, can induce the directional differentiation from BMMSCs to nerve cells. Salidroside, as the effective constituent of Rhodiola spp., has strong neuroprotection and anti-oxidation function. The subject was mainly to research the effect of salidroside on the directional differentiation from mouse BMMSCs to nerve cells, discussed the biological signal transmission mechanism of salidroside preliminarily, to lay the theoretical foundation of clinical application by using salidroside as the inducer of BMMSCs for repairing the nerve injure and provide scientific evidence for sifting more efficient inducers.【Method】The experiments were started after the cultured BMMSCs reached 80% confluency in vitro:â‘ The cells were assigned into the salidroside induction group, the positive control group and the blank control group. The related molecules of nerve cells were detected by immunocytochemistry, RT-PCR and Western blot method after the cells were impacted respectively.â‘¡The expression levels of neuron-specific molecules were detected after that the different blocking agents were applied to block the celluar Ca²⁺ signal pawthway respectively.â‘¢The expression of Wnt/β-catenin signal pathway related molecules were detected in the salidroside induction group, then the expression levels of neuron-specific molecules were detected after Wnt/β-catenin signal pathway blocked specially by Dkk-1.â‘£The expression levels of key molecules in Wnt/β-catenin signal pathway were analyzed in the different Ca²⁺ signal blocking groups.【Results】①The expression of the neuron-specific molecules including NSE,β-Tubulin III, Nurr1 mRNA could be found in the induction group, instead of the blank control group. GFAP mRNA as neurogliocyte-specific molecule was no expression in each group. The expression abundance of NSE protein increased obviously with the time expand, contrary to Nestin which is a marker of nerve stem cell could be found in the induction group, and both were no expression in the blank control group.â‘¡The expression of the neuron-specific molecules were conspicuously down-regulated when the extracellular Ca²⁺, L-type Ca²⁺ channel and IP3-Ca²⁺ signal pathway were blocked respectively, especially in IP3-Ca²⁺ signal pathway blocking group(p<0.01).â‘¢Cells with higher expression of Wnt/β-catenin signal pathway related molecules, such as Wnt3a, Lrp-6, Axin, GSK-3β,β-catenin mRNA andβ-catenin protein, could be found in the induction group, the blank control group had no expression except Axin and GSK-3βmRNA. Both of gene and protein ofβ-catenin were no expression, and the neuron-specific molecules were conspicuously down-regulated compared with the induction group when Wnt/β-catenin signal pathway were blocked by Dkk-1(p<0.01).â‘£The expression levels of the key molecules including Wnt3a, Lrp-6,β-catenin mRNA andβ-catenin protein in Wnt/β-catenin signal pathway were conspicuously down-regulated when the extracellular Ca²⁺ and L-type Ca²⁺ channel and IP3-Ca²⁺ signal pathway were blocked respectively, especially in IP3-Ca²⁺ signal pathway blocking group(p<0.01).【Conclusion】Salidroside can induce the differentiation from mouse BMMSCs to nerve cells. The biological effect is connected to Ca²⁺ signal and Wnt/β-catenin signal pathway, and IP3 dependent Ca²⁺/Wnt signal pathway play an important role on the signal transmission of salidroside in BMMSCs differentiation.