去乙酰化酶Sirt1对牛骨骼肌卫星细胞增殖和分化的影响  

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

作　　者：张正雨 杨培鸿 郭宏 李新 张林林 郭益文 胡德宝 丁向彬 ZHANG Zhengyu;YANG Peihong;GUO Hong;LI Xin;ZHANG Linlin;GUO Yiwen;HU Debao;DING Xiangbin(Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Breeding,School of Animal Science and Animal Medicine,Tianjin Agricultural University,Tianjin 300384,China)

机构地区：[1]天津农学院动物科学与动物医学学院,天津市农业动物繁育与健康养殖重点实验室,天津300384

出　　处：《畜牧兽医学报》2025年第2期603-610,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：天津市“131”创新型人才培养工程第一层次人选资助项目。

摘　　要：旨在探讨沉默信息调节因子2相关酶1(silent information regulator 2-related enzyme1,Sirt1)对牛骨骼肌卫星细胞(bovine skeletal muscle satellite cells,BSMSCs)增殖和成肌分化的影响。在本试验中,将小片段干扰RNA(siRNA)和靶向Sirt 1的过表达质粒转染入从3~4月龄胎牛臀腿肌剥离的原代卫星细胞中,在增殖和分化阶段的特定时间点收集RNA和蛋白质样品,每个处理设计3个生物学重复,使用实时定量PCR(qRT-PCR)以及蛋白质印记法(Western blotting)进行分析。此外,还进行了EdU染色试验以评估细胞的增殖情况。结果显示:在细胞分化期,40倍显微镜下观察,发现Sirt 1敲低导致肌管的数量、长度和直径显著增加。qRT-PCR和Western blotting证实,与对照组相比,分化标志物肌球蛋白重链(myosin heavy chains,MyHC)和肌细胞生成素(myogenin,MyoG)的mRNA水平显著上调(P<0.01),MyoG的蛋白表达水平也显著升高(P<0.05)。相反,Sirt 1过表达导致肌管的数量、长度和直径显著减少。Western blotting显示Sirt 1过表达组MyoG表达量显著低于对照组(P<0.01)。在BSMSCs增殖期,qRT-PCR、Western blotting和EdU检测表明,Sirt 1敲低和过表达对细胞增殖均无显著影响(P>0.05)。综上所述,Sirt 1敲减可有效促进细胞分化,而过表达可抑制该过程,但对增殖均无明显影响。这些发现证实了Sirt 1是作为调控BSMSCs成肌分化的重要因素,在协调成肌细胞分化和促进肌管形成中起关键作用。The aim of this study was to investigate the effect of silent information regulator 2-related enzyme1(Sirt 1)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 Sirt 1 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,Sirt 1 overexpression resulted in a significant reduction in the number,length,and diameter of muscle ducts.Western blotting showed that the expression of MyoG in Sirt 1 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 Sirt 1 knockdown and overexpression had no significant effects on cell proliferation(P>0.05).In summary,Sirt 1 knockdown can effectively promote cell differentiation,while overexpression can inhibit the process,but has no significant effect on proliferation.These findings confirm that Sirt 1 is an important factor regulating the myogenic differentiation of BSMSCs,and plays a key role in coordinating myoblast differentiation and promoting myotube formation.

关 键 词：SIRT1 牛骨骼肌卫星细胞 细胞增殖 成肌分化 

分 类 号：S823.2[农业科学—畜牧学]