Background:Skeletal Muscle Injury is Very Common in the Sports Medicine Field, and also the Important Factors Affecting the Sports Life of Athletes because Its Healing Quality is
Unstable. Myoblasts are the Precursor Cells to Rebuild Muscle Tissue after Trauma in Adult Skeletal Muscle. In Recent Years, Myoblasts Show Good Prospect in Clinical Application for the Self-Repair of
Injured Skeletal Muscle. However, Myoblasts are Prone to Spontaneous Differentiation and Difficult to Proliferation in Vitro, Which Greatly Reduced the Therapeutic Efficacy for Skeletal Muscle Injury
. So, Looking for more Effective Myoblast Regulation Factor in Vitro has a Great Significance in Both Theory and Valuable Application. Furthermore, the Effect of Salidroside on Skeletal Muscle Cells
Remains Unknown. C2C12 is a Murine Myogenic Cell Line that Proliferate Rapidly, and When Confluent and in the Presence of 2% HS (Horse Serum), They Begin to Fuse and Differentiate into Myotubes. In
the Present Work, We Show that Salidroside Inhibits in Vitro Myogenesis though Activating P-smad3 Expression in a Dose-dependent Manner in C2C12 Cells.
Methods:1. Observe the Morphology of Differential Myoblast C2C12 Which were Treated With 0, 30, 50ug/ml Salidroside to Induce Differentiation by Phase Micrographs; 2.Detect the Cell
Fusion Index Affected by Salidroside Using Immunofluorescence;3.Detect the Expressions of Myogenic Differentiation-Specific mRNAs and Proteins Affected By Salidroside Using Western Blotting and Real
Time-PCR.4. Detect TGF-Β/Smad3 Pathway Protein Expression in Differential C2C12 Cell Treated with 0, 30, 50ug/ml Sal Using Western Blotting. 5. Inhibit Smad3 Phosphorylation with Smad3
Inhibitors(SIS3), then to Repeat Method 1-3 to Detect how SIS3 Impact the Effect of Salidroside in Method 1-3.
Results: Treated with 50ug/ml Salidroside: 1. Salidroside Inhibit C2C12 Fusion and Myotube Form in Morphological By Phase Micrographs; 2. Myosin-Neonatal Positive Area and Myogenin
Positive Nuclei Decreased Significantly by Immunofluorescence（P＜0.05）, Which Inhibited Myotube Form Notably; 3. The Expressions of MyoD and Myogenin in mRNA and Protein both Decreased Notably by
Real-Time PCR and Western Blotting（P＜0.05）, Which Inhibited Cell Fusion and Myotube Form during C2C12 Differentiation. 4.Salidroside Inhibit C2C12 Differentiation Associated with Increases in
Smad2/Smad3 Phosphorylation, Especially P-smad3（P＜0.01）; 5. After Selectively Abrogated Smad3 Phosphorylation, the Effect of Salidroside in Result1-3 Invalided;
The Results Treated with 30ug/ml Salidroside is Insignificant （P＞0.05）.
Conclusions:1. Salidroside Inhibit Myogenic Differentiation of C2C12 Cell Significantly, and Promote Cell Proliferation, to Add Satellite Cells Band Reserve . 2. Salidroside Inhibits
Myogenic Differentiation in a Dose-dependent Manner, 50ug/ml Salidroside is the Best. 3. Salidroside Inhibits Myogenic Differentiation by Activating Smad3 Phosphorylation.