去乙酰化酶Sirt1对牛骨骼肌卫星细胞增殖和分化的影响

doi:10.11843/j.issn.0366-6964.2025.02.012

Abstract

Abstract:

The aim of this study was to investigate the effect of silent information regulator 2-related enzyme1 (Sirt1) on proliferation and myogenic differentiation of BSMSCs. In this experiment, small fragments of interfering RNA (siRNA) and overexpressed plasmid targeting Sirt1 were transfected into primary satellite cells stripped from the thigh muscle of 3-4-month-old fetal cattle. RNA and protein samples were collected at specific time points during the proliferation and differentiation stages, with 3 biological replicates per treatment. Real-time quantitative PCR (qRT-PCR) and Western blotting were used for analysis. In addition, EdU staining experiments were performed to evaluate cell proliferation.The results showed that the number, length, and diameter of muscle ducts were significantly increased by Sirt1 knockdown under a 40-fold microscope during cell differentiation. qRT-PCR and Western blotting confirmed that compared with the control group, mRNA levels of differentiation markers myosin heavy chains (MyHC) and myogenin (MyoG) were significantly up-regulated after interfering Sitr1 (P < 0.01). The protein expression level of MyoG was also significantly increased (P < 0.05) after interfering Sitr1. Conversely, Sirt1 overexpression resulted in a significant reduction in the number, length, and diameter of muscle ducts. Western blotting showed that the expression of MyoG in Sirt1 overexpression group was significantly lower than that in control group (P < 0.01). At the proliferation stage of BSMSCs, qRT-PCR, Western blotting and EdU tests showed that Sirt1 knockdown and overexpression had no significant effects on cell proliferation (P>0.05). In summary, Sirt1 knockdown can effectively promote cell differentiation, while overexpression can inhibit the process, but has no significant effect on proliferation. These findings confirm that Sirt1 is an important factor regulating the myogenic differentiation of BSMSCs, and plays a key role in coordinating myoblast differentiation and promoting myotube formation.

Key words: Sirt1, bovine skeletal muscle satellite cells, cell proliferation, myogenic differentiation

CLC Number:

S823.2

ZHANG Zhengyu, YANG Peihong, GUO Hong, LI Xin, ZHANG Linlin, GUO Yiwen, HU Debao, DING Xiangbin. Effects of Sirt1 Deacetylase on Proliferation and Differentiation of Bovine Skeletal Muscle Satellite Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(2): 603-610.

Figures/Tables 7

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引物名称 Primers name	基因ID GenBank ID	引物序列(5′→3′) Sequence	产物大小/bp Products size
Sirt1 forward	NM_001192980.3	CATTTTCCATGGCGCTGAGG	104
Sirt1 reverse	NM_001192980.3	AACTTGGACTCTGGCACGTT	104
GAPDH forward	NM_001034034.2	CCGCATCCCTGAGACAAGAT	114
GAPDH reverse	NM_001034034.2	ATTGATGGCGACGATGTCCA	114
Pax7 forward	XM_015460690.3	CGATTAGCCGAGTGCTCAGA	137
Pax7 reverse	XM_015460690.3	GTCCAGACGGTTCCCTTTGT	137
Ki67 forward	NM_001034354.2	CCTAAACCCGCAGGAGGATG	78
Ki67 reverse	NM_001034354.2	TTCTCCTGTTGCTTGGTCGC	78
MyoG forward	NM_001111325.1	GTGGGCGTGTAAGGTGTGTA	91
MyoG reverse	NM_001111325.1	CACCTTCTTGAGTCTGCGCT	91
MyHC forward	NM_174117.1	TCAAGGTTGCATCTCCAAGGC	131
MyHC reverse	NM_174117.1	GCGCTCCTTTTCAGACTTTCG	131

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Effects of Sirt1 Deacetylase on Proliferation and Differentiation of Bovine Skeletal Muscle Satellite Cells

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