The Regulation Of MGF On The Migration Of Muscle Satellite Cells And The Role Of ERK In It

Objective Skeletal muscle damage phenomenon is very common in People’s Daily physical activity, but its healing effects are not perfectly good, the reason was that the self-healing of skeletal muscle has its limitations, so how to make the skeletal muscle heal more quickly with high quality,has been into a hot point in sports medicine and molecular biology. In skeletal muscle damage, muscle satellite cells start to play a decisive role, they pass through their own behavior, such as proliferation, migration affect the regeneration of skeletal muscle. Previous research on MGF migration pathways are relatively rare. This study using in vitro culture of skeletal muscle satellite cells, combination of PD98059(Erk1/2) inhibitors under MGF intervention,exploresignaling pathways of MGF on the migration of muscle satellite cells.Materials and Methods We extracted hind limbsgastrocnemius and soleus from SD rats(male)under sterile conditions.After fully cut up,we usedthe two-step enzyme digestion method digesting muscle cells release,and purified muscle satellitecells through twice differential adhesion.Then the viability rate and purity of muscle satellite cells were identified by trypan blue and a-sarcometric actin respectively.We usedthe three generations of cells with 45 ng/ml of synthetic MGF intervention, the experimental group: group C, MGF group, the MGF + PD group,each group respectively intervention 24, 48, 72 and 96 h.Using CCK-8 experiment to measure muscle satellite cell’s proliferation ability,the cell migration rate was measured by Transwell assay and the inhibitors effects were detected by Western Bolt.Results1. Trypan blue staining results: Assessment of cell viability showed that the live SC proportion was96.6%.2. The immunocytochemistry of a-sarcometric actin results: Cells were detected in a tan colored, were positive, showing they were muscle satellite cells.3. CCK-8 results: Comparison group C and MGF group’s data showed that MGF group at 24, 48, 72, 96 h the increment of the OD were significantly higher than group C(P < 0.01).4. Transwell results:1) MGF group: 48, 72 h group’s OD of skeletal muscle satellite cell migration was significant higher than 24 h group(P < 0.01), 96 h group’s OD was also significantly higher than 24 h group(P < 0.05).2) Compare MGF group and MGF + PD group,showed that in 24, 48, 72, 96 h group, MGF group and MGF + PD group’s OD of the skeletal muscle satellite cell migration group had no significant differences(P > 0.05).5. Western Bolt results:Using Western Blot test P-Erk1/2 in the expression of MGF and MGF + PD group,the results showed that MGF + PD group at 24, 48, 72 h, P-Erk1/2 expression quantity was significantly lower than the MGF group(P < 0.01), MGF + PD group at96 h, P-Erk1/2 expression quantity was lower than that of MGF group(P < 0.05).Conclusions1. Exogenous MGF can promote the proliferation of skeletal muscle satellite cells,and have effects on the proliferation of timeliness, the best time for its proliferations72 h, within 24-72 h, promote cell proliferation effect showed a trend of rising and within 72-96 h, promote cell proliferation effect showed a trend of falling.2. Exogenous MGF can promote skeletal muscle satellite cell migration, and skeletal muscle satellite cell’s proliferation might affect their migration effect.3. ERK inhibitors(PD) on skeletal muscle satellite cell migration effect is not significant,so MAPK/ERK1/2 signaling pathways may not affects skeletal muscle satellite cell’s migration.