Naringenin repairs skeletal muscle injury by regulating polarization of macrophages and proliferation of muscle satellite cells

doi:10.12307/2023.058

Abstract

Abstract: BACKGROUND: Studies have shown that the precise regulation of macrophage polarization is extremely important for tissue repair, the proliferation and differentiation of muscle satellite cells during the repair of skeletal muscle injury. Naringenin can improve excessive fibrosis during skeletal muscle injury repair.
OBJECTIVE: To explore the effect and mechanism of macrophage polarization in skeletal muscle repair, thereby providing a theoretical basis for the treatment of skeletal muscle injury.
METHODS: Eighty 10-week-old Sprague-Dawley rats of specific pathogen-free grade were randomly divided into two groups (n=40). A skeletal muscle injury model was constructed by weight drop method. The experimental group was intraperitoneally injected with naringenin (2 μg/g) for 14 days and the control group was injected with the same amount of 1% dimethyl sulfoxide. Samples of gastrocnemius muscle were collected from 6 rats in each group at 12 hours and 1, 3, 5, 7, and 14 days after injury. Masson and hematoxylin-eosin staining were used to observe the degree of skeletal muscle tissue fibrosis, and enzyme-linked immunosorbent assay was used to detect the expression of inflammatory factors interleukin-4, interleukin-13, interferon-α, and interferon-γ. The mRNA expressions of pro-fibrotic genes (type I and III collagens) and myogenic differentiation antigen (MyoD) were detected by real-time fluorescence quantitative PCR, Polarization degree of macrophages was detected by flow cytometry. Proliferation of muscle satellite cells was observed by immunofluorescence (Pax7, MyoD) staining.
RESULTS AND CONCLUSION: Histological observations showed that the fibrosis area ratio of the experimental group was lower than that of the control group at 5, 7, and 14 days after injury (P < 0.05), and the size of regenerated muscle fibers was significantly larger than that of the control group (P < 0.05). The ELISA results showed that compared with the control group, the expression of interleukin-4 at 7 and 14 days after injury and the expression of interleukin-13 and interferon-α at 5, 7, and 14 days after injury increased significantly in the experimental group (P < 0.05), while the expression of interferon-γ declined. Real-time fluorescence quantitative PCR results revealed that compared with the control group, the relative expression of type I and III collagens decreased significantly in the experimental group at 3, 5, 7, and 14 days after injury (P < 0.05), while the relative expression of MyoD increased significantly at 3, 5, and 7 days after injury (P < 0.05). Flow cytometry results showed that the number of M1-type macrophages in the experimental group was lower than that of the control group at 3, 5, 7, and 14 days after injury, while the number of M2-type macrophages was significantly higher than that of the control group (P < 0.05). Immunofluorescence staining results showed that the proliferation of muscle satellite cells in the experimental group was significantly higher than that in the control group at 3 days after injury (P < 0.05). To conclude, naringenin can treat skeletal muscle injury by regulating the M2-type polarization of macrophages to promote the proliferation of muscle satellite cells.

Key words: skeletal muscle, injury, naringin, fibrosis, macrophage polarization, rat

CLC Number:

Xu Mingkui, Xu Riming, Lin Yewu, Luo Yuantai, Zhang Xihui, Zhou Li, Yuan Shiguo. Naringenin repairs skeletal muscle injury by regulating polarization of macrophages and proliferation of muscle satellite cells[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(14): 2133-2138.

Figures/Tables 9

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