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2025年5月1日 星期四
农业生物技术学报  2022, Vol. 30 Issue (4): 695-703    DOI: 10.3969/j.issn.1674-7968.2022.04.008
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
烟酰胺单核苷酸(NMN)对小鼠C2C12细胞成肌分化的影响
沈龙仙, 王丽婷, 汪涵, 杨松柏, 李向臣, 赵阿勇*, 周晓龙*
浙江农林大学 动物科技学院/动物医学院,临安 311300
Effects of Nicotinamide Mononucleotide (NMN) on Myogenic Differentiation of Mouse (Mus musculus) C2C12 Cells
SHEN Long-Xian, WANG Li-Ting, WANG Han, YANG Song-Bai, LI Xiang-Chen, ZHAO A-Yong*, ZHOU Xiao-Long*
College of Animal Science and Technology/College of Veterinary Medicine, Zhejiang A&F University, Lin'an 311300, China
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摘要 烟酰胺单核苷酸(nicotinamide mononucleotide, NMN)具备多种生物学功能,对心脑疾病、老年退行性疾病、神经退行性疾病、抗衰老等具有治疗作用。经NMN处理后的骨骼肌对衰老相关的转录变化有显著的预防作用,但NMN对骨骼肌增殖分化有何作用尚不明确。为深入了解NMN对细胞成肌分化的影响、丰富肌肉生长发育的机制,本研究以小鼠(Mus musculus)成肌细胞(mouse myoblasts) C2C12为模型,将不同浓度的NMN (0, 0.1, 1和2 μg/mL)应用于2%马血清(horse serum, HS)诱导分化的C2C12。采用CCK-8 (Cell Counting Kit-8)法检测NMN处理后C2C12细胞的活力;通过2',7'-二氯荧光黄双乙酸盐(2',7'-dichlorodi-hydrofluorescein diacetate, DCFH-DA)和四氯四乙基苯并咪唑碳菁碘(5,5',6,6'-tetrechloro-1,1',3,3'-tetraethyl benzimidazol carbocyanine iodide, JC-1)两种荧光探针,分别对细胞进行染色,检测NMN对C2C12细胞线粒体膜电位和活性氧(reactive oxygen species, ROS)的影响;利用Western blot及免疫荧光技术检测肌细胞分化标记因子肌生成素(myogenin, MyoG)、肌分化因子1 (myogenic differentiation 1, MyoD1)表达和肌球蛋白重链(myosin heavy chain, MYH)的蛋白表达水平。结果显示,0、0.1、1和2 μg/mL的NMN对C2C12细胞没有毒性作用,并且一定浓度的NMN在一定时间内可以促进细胞活力,提高线粒体膜电位,上调成肌分化相关蛋白MyoG、MyoD1和MYH的表达。上述研究结果表明,一定浓度的NMN在一定时间内可以促进C2C12细胞成肌分化。本研究丰富了NMN的生理作用,为后续NMN在畜牧产业的应用提供新的参考基础。
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沈龙仙
王丽婷
汪涵
杨松柏
李向臣
赵阿勇
周晓龙
关键词 烟酰胺单核苷酸(NMN)C2C12细胞成肌分化肌肉特异性基因    
Abstract:Nicotinamide mononucleotide (NMN) has a variety of biological functions, which are useful for heart and brain diseases, senile degenerative diseases, neurodegenerative diseases, and anti-aging. The skeletal muscle treated with NMN has significantly preventive effect on the transcriptional changes related to aging, but the effect of NMN on the proliferation and differentiation of skeletal muscle is still unclear. In order to understand the effect of NMN on cell myoblast differentiation and enrich the mechanism of muscle growth and development, this study used mouse ((Mus musculus) myoblasts (C2C12) as a model, and different concentrations of NMN (0, 0.1, 1 and 2 μg/mL) was applied to 2% horse serum (HS) to induce differentiation of C2C12. The Cell Counting Kit-8 method (CCK-8 method) was used to detect the viability of C2C12 cells after NMN treatment; The cells were stained by 2 fluorescent probes, 2',7'-dichlorodi hydrofluorescein diacetate (DCFH-DA) and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl benzimidazole carbocyanine iodide (JC-1), respectively, the effects of NMN on mitochondrial membrane potential and reactive oxygen species (ROS) of C2C12 cells were detected; Western blot (WB) and immunofluorescence technique were used to detect the expression of myogenin (MyoG), myogenic differentiation 1 (MyoD1) and myosin heavy chain (MYH). The results showed that 0, 0.1, 1 and 2 μg/mL NMN had no toxic effect on C2C12 cells, and a certain concentration of NMN could promote cell viability, increased mitochondrial membrane potential, and up-regulated the expression of myogenic differentiation related proteins MyoG, MyoD1 and MYH within a certain period of time. The above research results indicated that a certain concentration of NMN could promote the myogenic differentiation of C2C12 cells within a certain period of time. This research enriches the physiological role of NMN by exploring the role of NMN in cell myogenic differentiation, and provides a new reference basis for the subsequent application of NMN in the animal husbandry industry.
Key wordsNicotinamide mononucleotide (NMN)    C2C12 cell line    Myoblast differentiation    Muscle specific gene
收稿日期: 2021-04-07     
ZTFLH:  S835  
基金资助:国家自然科学基金(31872397); 浙江省自然科学基金(LY19C170003)
通讯作者: *zhouxiaolong@zafu.edu.cn;zay503@zafu.edu.cn   
引用本文:   
沈龙仙, 王丽婷, 汪涵, 杨松柏, 李向臣, 赵阿勇, 周晓龙. 烟酰胺单核苷酸(NMN)对小鼠C2C12细胞成肌分化的影响[J]. 农业生物技术学报, 2022, 30(4): 695-703.
SHEN Long-Xian, WANG Li-Ting, WANG Han, YANG Song-Bai, LI Xiang-Chen, ZHAO A-Yong, ZHOU Xiao-Long. Effects of Nicotinamide Mononucleotide (NMN) on Myogenic Differentiation of Mouse (Mus musculus) C2C12 Cells. 农业生物技术学报, 2022, 30(4): 695-703.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2022.04.008     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2022/V30/I4/695
 
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