Directional differentiation of mouse mesenchymal stem cells into neuronal cells induced by salidroside via Notch and BMP signal pathway / 中草药

ABSTRACT

Objective: To study the molecule mechanism of salidroside inducing mesenchymal stem cells (MSCs) to directionally differentiate into neuronal cells via bone morphogenetic protein (BMP) and Notch signal pathways. Methods: Experiments were divided into control, induced, and blocked groups. The technologies, such as immunofluorescence, real-time PCR, and Western blotting were used to analyze the effect of salidroside on cellular proliferation, morphosis, and BMP and Notch signal pathways. Results: The immunofluorescence results showed that salidroside could affect cellular proliferation and induce MSCs to form the morphosis of neuronal cells. The positive rate of Notch1 and Jadge1 was significantly decrease to compare with the control (P < 0.05), real-time PCR results indicated that mRNA expression of Notch1 and Hes1 was obviously down-upregulated when treated with salidroside for 12-72 h (P < 0.05). However, Notch signal pathway was blocked with DAPT, a special inhibitor of Notch, the marker molecules of neuronal cells expression, such as neuron-specific enolase (NSE), microtubule associated protein 2 (MAP2), and β-tubulin III, were significantly increased when cells were treated with salisroside (P < 0.05). The mRNA levels of Smad5 and Smad8 were up-regulated when cells were treated with salidroside for 12 h, expression of Smad1/5/8 protein was increased at 12 and 24 h. When BMP signal pathway was blocked with Noggin, a special inhibitor of BMP, NSE, MAP2, and β-tubulin III mRNA expression was decreased to compare with the salidroside induced group (P < 0.05); When Notch and BMP signal pathways were simultaneously blocked with DAPT and Noggin, MAP2 and β-tubulin III mRNA expression was increased more obviously than that of the blocked with Noggin. Meanwhile the expression of NSE and β-tubulin III protein was also up-regulated (P < 0.05). Conclusion: Salidroside promotes neuron-like differentiation of MSCs by negatively regulating the Notch pathway and activating BMP signal pathway, it plays a vital role for salidroside to inhibit Notch pathway on affecting MSCs differentiation.