干扰核心蛋白聚糖对牛骨骼肌卫星细胞增殖分化的影响

doi:10.16431/j.cnki.1671-7236.2021.01.023

Abstract

Abstract: In order to explore the mechanism of myostatin (MSTN) on the growth and development of bovine skeletal muscle,this study aimed at the expression of decorin (DCN) with large fold difference obtained by quantitative proteomic and phosphoproteomic screening of muscle tissues of MSTN^+/- Mongolian cattle and wild Mongolian calf gluteal muscle,and isolated and cultured bovine skeletal muscle satellite cells in the pre-laboratory.The model of induced myogenic differentiation in vitro was established as the object.Through screening the interference effects of three DCN siRNAs designed and synthesized,the most significant interference effect of si-DCN-2 (si-DCN) was transfected into bovine skeletal muscle satellite cells.Real-time fluorescence quantitative PCR and Western blotting were used to detect the changes in the expression of the proliferation markers Pax7 and MyoD in proliferative phase (GM) bovine skeletal muscle satellite cells and the effects of interfering DCN on cell proliferation was detected using EdU staining.In vitro myogenic differentiation of bovine skeletal muscle satellite cells transfected with DCN siRNA was performed.Myotube formation on the third day of differentiation (DM3) of bovine skeletal muscle satellite cells was observed by microscopy,and Real-time fluorescence quantitative PCR and Western blotting were used to detecte the expression changes of the differentiation markers MyoG and MyHC at the mRNA level and protein level,and myotube MyHC at DM3 stage was stained by immunofluorescence to study the effect of interfering with DCN on cell differentiation.The results showed that after interfering with DCN expression,the mRNA and protein levels of Pax7 and MyoD in bovine skeletal muscle satellite cells in the proliferative phase were significantly or extremely significantly upregulated (P<0.05;P<0.01),and the rate of EdU-positive cells was significantly increased (P<0.05),indicating that interfering with DCN expression significantly promoted the proliferation of bovine skeletal muscle satellite cells.After interfering with the expression of DCN,the diameter of myotubes induced by bovine skeletal muscle satellite cells on the third day of differentiation showed an increasing trend.The expression of MyoG,a marker of myogenic differentiation,was extremely significantly and significantly higher than that of the control group at the mRNA and protein levels respectively (P<0.01;P<0.05).MyHC was significantly lower at the mRNA level (P<0.05),but significantly higher at the protein level (P<0.01).Immunofluorescence results showed that the myotube fusion index was significantly higher after downregulation of DCN than that of the control group (P<0.05),indicating that interfering with DCN expression can promote the myogenic differentiation process of bovine skeletal muscle satellite cells.The results of this study showed that interfering with DCN could significantly promote the proliferation and myogenic differentiation of bovine skeletal muscle satellite cells.The results laid a foundation for further research on the regulatory mechanism of MSTN on myogenic differentiation of bovine skeletal muscle satellite cells.

Key words: decorin (DCN); bovine; skeletal muscle satellite cells; proliferation; myogenic differentiation

CLC Number:

S813.3

SHENG Hui, GUO Yiwen, ZHANG Linlin, MIAO Manning, LI Xin, DING Xiangbin, GUO Hong. Effects of Interfering with Decorin on Proliferation and Differentiation of Bovine Skeletal Muscle Satellite Cells[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(1): 209-217.

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Effects of Interfering with Decorin on Proliferation and Differentiation of Bovine Skeletal Muscle Satellite Cells

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