Platelet-derived growth factor-BB promotes proliferation, differentiation and migration of skeletal muscle myoblast

doi:10.3969/j.issn.2095-4344.2171

Abstract

Abstract: BACKGROUND: Skeletal muscle myoblasts differentiate and fuse to form polynuclear muscle tubes and muscle fibers to complete the repair of muscle injury when skeletal muscles were injured. However, the repair process is slow and incomplete. Platelet-derived growth factor-BB can stimulate the proliferation, differentiation and migration of a variety of tissue cells, and plays an important role in the process of tissue repair after various injuries.
OBJECTIVE: To explore the effects of different concentrations of platelet-derived growth factor-BB on proliferation, differentiation and migration of skeletal muscle myoblast C2C12 cells and the action mechanism.
METHODS: The mouse skeletal muscle myoblast C2C12 cells were cultured with platelet-derived growth factor-BB 0, 5, 10, 20 and 40 μg/L and platelet-derived growth factor receptor inhibitor imatinib. The expression of platelet-derived growth factor receptor in C2C12 cells was detected by immunocytochemistry and western blot assay. After 1, 2, 3, 4, and 5 days of culture, CCK8 method was used to detect cell proliferation. After 4 days of induction and differentiation culture, the formation of myotubes in each group was observed by light microscope. The expression of myosin heavy chain and MyoG Gene was observed by immunofluorescence and western blot assay. After 48 hours of culture, Transwell method was used to detect cell migration.
RESULTS AND CONCLUSION: (1) Immunofluorescence chemistry and western blot assay indicated that platelet-derived growth factor receptor expression could be detected in C2C12 cells, and semi-quantitative statistical analysis showed no significant difference in platelet-derived growth factor receptor expression between groups with different platelet-derived growth factor-BB concentrations (P > 0.05). (2) CCK8 assay demonstrated that the proliferation of C2C12 cells showed no significant change in platelet-derived growth factor-BB groups compared with the control group (platelet-derived growth factor-BB 0 μg/L group). (3) Immunofluorescence chemistry showed that compared with control group, number of myosin heavy chain positive cells increased in platelet-derived growth factor-BB groups; 40 μg/L platelet-derived growth factor-BB concentration got the highest; the mature muscle tubes up to (27.00±0.76) per field of vision, and MyoG expression populations was most obviously compared with the control group (P < 0.05). (4) Transwell results showed that compared with the control group, the migration number of C2C12 cells in the platelet-derived growth factor-BB group increased, and the migration number in the 40 μg/L group was up to 144.00±13.03 (P < 0.05). (5) It is concluded that platelet-derived growth factor-BB promoted the migration, differentiation and myotube formation of C2C12 cells, and the pro-differentiation mechanism is related to its enhanced binding with platelet-derived growth factor receptor, so as to improve the expression of differentiation related gene MyoG.

Key words: skeletal muscle, myoblasts, platelet-derived growth factor-BB, cell proliferation, cell differentiation, cell migration, protein, myosin heavy chain

CLC Number:

Li Cai, Zhao Ting, Tan Ge, Zheng Yulin, Zhang Ruonan, Wu Yan, Tang Junming. Platelet-derived growth factor-BB promotes proliferation, differentiation and migration of skeletal muscle myoblast[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1050-1055.

Figures/Tables 5

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