抑制小鼠成肌细胞O-GlcNAc对其细胞周期和细胞分化的影响  

Effects of O-GlcNAc inhibition on cell cycle and differentiation of mouse C2C12 cells

作　　者：史宏昭 姚睿智[3] SHI Hongzhao;YAO Ruizhi(College of Animal Engineering,Yangling Vocational and Technical College,Yangling 712100,China;College of Veterinary Medicine,Northwest A&F University,Yangling 712100,China;College of Animal Science and Technology,Inner Mongolia Minzu University,Tongliao 028011,China)

机构地区：[1]杨凌职业技术学院动物工程学院,陕西杨凌712100 [2]西北农林科技大学动物医学院,陕西杨凌712100 [3]内蒙古民族大学动物科技学院,内蒙古通辽028000

出　　处：《黑龙江动物繁殖》2024年第5期7-12,19,共7页Heilongjiang journal of animal reproduction

基　　金：杨凌职业技术学院院自然科学项目(ZK22-73);内蒙古自治区自然科学基金项目(2022QN03028);内蒙古民族大学青年教师科研创新能力提升计划项目(GXKY23Z001)。

摘　　要：成肌细胞的高效增殖分化对于畜牧业肉产品生产具有重要意义。试验旨在通过特异性抑制剂OSMI-1抑制氧连接的β-N-乙酰葡萄糖胺(O-linked N-acetylglucosamine glycosylation,O-GlcNAc)修饰检测小鼠成肌细胞系(C2C12)细胞周期和分化。试验以小鼠C2C12细胞为试验对象,设置空白组、DMSO组和OSMI-1/DMSO组,采用Western Blot确定OSMI-1对O-GlcNAc修饰抑制,采用流式细胞术检测细胞周期变化,使用免疫荧光检测肌管分化标志(myosin heavy chain,Myhc)和qRT-PCR检测分化因子(myogenic differentiation,Myod)及Myhc基因表达水平,分析O-GlcNAc对C2C12肌管分化的影响。结果表明:与空白组和DMSO组相比,OSMI-1/DMSO组24 h和48 h显著抑制了C2C12的O-GlcNAc修饰水平,其细胞周期G2细胞比例显著下降(P<0.05);分化过程中抑制O-GlcNAc修饰水平,免疫荧光显示C2C12分化标志Myhc表达减少,Myod和Myhc的转录和翻译水平显著下降(P<0.05)。说明低水平O-GlcNAc修饰抑制C2C12细胞的细胞周期和细胞分化。The efficient proliferation and differentiation of myoblasts are crucial for the production of livestock meat products.In this experiment,the specific inhibitor OSMI-1 was employed to inhibit O-linkedβ-N-acetylglucosamine(O-GlcNAc)glycosylation to observe the changes in the cell cycle and differentiation of the mouse myoblast cell line(C2C12).Mouse C2C12 cells were used as experimental subjects,with a blank group,a DMSO group,and an OSMI-1/DMSO group established.Western Blot was utilized to determine the inhibition of O-GlcNAc modification by OSMI-1;flow cytometry was used to detect changes in the cell cycle;immunofluorescence was used to detect the differentiation marker(myosin heavy chain,Myhc),and qRT-PCR was used to detect the expression levels of differentiation factors(myogenic differentiation,Myod)and Myhc genes,to analyze the effect of O-GlcNAc on C2C12 myotube differentiation.The results indicated that the O-GlcNAc modification level of C2C12 was significantly inhibited by the OSMI-1/DMSO group at 24 h and 48 h,and the proportion of G2 cells in the cell cycle was significantly reduced(P<0.05).During the differentiation process,the inhibition of O-GlcNAc modification resulted in a decrease in the expression of the differentiation marker Myhc,as well as a decrease in the expression of Myod and Myhc.

关 键 词：小鼠成肌细胞系(C2C12) β-N-乙酰葡萄糖胺(O-GlcNAc) OSMI-1 细胞周期 分化 

分 类 号：Q254[生物学—细胞生物学]