褪黑素对脂多糖刺激的滩羊骨骼肌卫星细胞炎性因子表达的影响

doi:10.16431/j.cnki.1671-7236.2021.09.029

Abstract

Abstract: The aim of this study was to investigate the effect of melatonin (MLT) on lipopolysaccharide (LPS)-induced inflammatory response in skeletal muscle satellite cells in Tan sheep. In this study, the 30-day-old fetus of Tan sheep was selected. The skeletal muscle satellite cells were isolated by collagenase Ⅳ, and cultured in vitro, the corresponding inductive reagent was added to induce differentiation of skeletal muscle satellite cells, and the expression of surface markers CD29, CD44, CD73 and Vimentin were analyzed by immunofluorescence. The skeletal muscle satellite cells were cultured at different doses (0, 5, 8 and 10 μg/mL) of LPS in the medium without fetal bovine serum, and the mRNA levels of inflammatory related factors such as interleukin-6 (IL-6), IL-8 and tumor necrosis factor-α (TNF-α) were detected by Real-time quantitative PCR (qRT-PCR), and the optimal concentration of LPS was selected. Skeletal muscle satellite cells were co-cultured with different concentrations 0.1 and 0.5 μmol/L of MLT and LPS of optimal concentration, and the mRNA levels of IL-6, IL-8 and interferon-γ (IFN-γ) were detected by qRT-PCR. The immunofluorescence results showed that the expression of the surface markers CD29, CD44, CD73 and Vimentin of skeletal muscle satellite cells were positive, and the cells had the properties of myogenic, lipogeneic and osteogenic differentiation. qRT-PCR results showed that different concentrations of LPS treatments could significantly increase the genes expression of inflammatory related factor when compared with control group (P<0.05), and the mRNA expression of inflammatory factors was the strongest when treated with 8 μg/mL LPS. When the cells were co-cultured by MLT and LPS, 0.5 μmol/L MLT could significantly decrease the expression of IL-6 induced by LPS when compared with LPS group (P<0.05). In conclusion, MLT could alleviate LPS-induced inflammatory response in skeletal muscle satellite cells in Tan sheep. The results laid a foundation for further research on the regulatory mechanism of MLT alleviating the inflammatory response of skeletal muscle satellite cells induced by LPS.

Key words: Tan sheep; skeletal muscle satellite cells; lipopolysaccharide (LPS); melatonin (MLT); inflammation

CLC Number:

S826

MA Sijia, LIU Yuan, WANG Xuzhong, LI Tingting, DUAN Xing, YANG Songbai, SONG Dan, LI Xiangchen. Effects of Melatonin on the Expression of Inflammatory Factors in Skeletal Muscle Satellite Cells Stimulated by LPS in Tan Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(9): 3378-3386.

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Effects of Melatonin on the Expression of Inflammatory Factors in Skeletal Muscle Satellite Cells Stimulated by LPS in Tan Sheep

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