摘要
为探究二甲双胍对牛骨骼肌卫星细胞增殖和分化的影响,本研究将体外培养的牛骨骼肌卫星细胞分别用0(对照组)、1、2、4 mmol/L二甲双胍进行处理,采用CCK-8法筛选出二甲双胍作用于牛骨骼肌卫星细胞的最适浓度,接着通过EdU染色法检测二甲双胍处理牛骨骼肌卫星细胞后对其增殖的影响,然后对二甲双胍处理的牛骨骼肌卫星细胞进行体外成肌诱导分化,通过显微镜观察牛骨骼肌卫星细胞分化时期的细胞状态,然后利用Western blotting技术检测牛骨骼肌卫星细胞的分化标志因子肌球蛋白重链(MyHC)、肌细胞生成素(MyoG)在分化24、48和72 h的表达情况。结果表明,二甲双胍作用于牛骨骼肌卫星细胞的最适浓度为2 mmol/L。2 mmol/L二甲双胍处理牛骨骼肌卫星细胞后,其细胞增殖率显著降低(P<0.05),说明二甲双胍可以抑制牛骨骼肌卫星细胞的增殖;牛骨骼肌卫星细胞诱导分化后形成的肌管数量和直径均呈现减少趋势,牛骨骼肌卫星细胞成肌分化标志因子MyHC、MyoG在分化24、48和72 h的表达均显著低于0 mmol/L(对照)组(P<0.05),说明2 mmol/L二甲双胍能够抑制牛骨骼肌卫星细胞的成肌分化过程。研究结果表明,二甲双胍可以显著抑制牛骨骼肌卫星细胞的增殖及成肌分化过程。该研究为二甲双胍在肌肉发育调控及肌损伤修复方面的应用提供一定的理论依据。
In order to explore the effects of metformin on the proliferation and differentiation of bovine skeletal muscle satellite cells,bovine skeletal muscle satellite cells cultured in vitro were treated with 0(control group),1,2 and 4 mmol/L metformin,and using CCK-8 method to screen the optimal concentration of metformin on bovine skeletal muscle satellite cells.The effect of metformin treatment on the proliferation of bovine skeletal muscle satellite cells was detected by EdU staining method,and the metformin-treated bovine skeletal muscle satellite cells were myogenically induced and differentiated in vitro,and the cell status of bovine skeletal muscle satellite cells during the differentiation period was observed.Then Western blotting was used to detect the expression of the differentiation markers myosin heavy chain(MyHC)and myogenin(MyoG)of bovine skeletal muscle satellite cells at 24,48 and 72 h of differentiation.The results showed that the optimal concentration of metformin on bovine skeletal muscle satellite cells was 2 mmol/L.After treating bovine skeletal muscle satellite cells with 2 mmol/L metformin,the cell proliferation rate were significantly reduced(P<0.05),indicating that metformin could inhibit the proliferation of bovine skeletal muscle satellite cells.The number and diameter of myotubes formed by bovine skeletal muscle satellite cells after induced differentiation showed a decreasing trend,and the expressions of myogenic differentiation markers MyHC and MyoG of bovine skeletal muscle satellite cells were significantly lower than 0 mmol/L(control)group(P<0.05)at 24,48 and 72 h after differentiation,indicating that 2 mmol/L metformin could inhibit the myogenic differentiation process of bovine skeletal muscle satellite cells.The results of this study showed that metformin could significantly inhibit the proliferation and myogenic differentiation of bovine skeletal muscle satellite cells,providing a theoretical basis for the application of metformin in muscle development regulation and muscle injury
作者
刘秋华
朱菲菲
王轶敏
张林林
李新
郭益文
郭宏
丁向彬
LIU Qiuhua;ZHU Feifei;WANG Yimin;ZHANG Linlin;LI Xin;GUO Yiwen;GUO Hong;DING Xiangbin(Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry,College of Animal Science and Veterinary Medicine,Tianjin Agricultural University,Tianjin 300384,China)
出处
《中国畜牧兽医》
CAS
北大核心
2021年第9期3354-3360,共7页
China Animal Husbandry & Veterinary Medicine
基金
天津市自然科学基金项目(18JCYBJC29700)
天津市“131”创新型人才培养工程第一层次人选资助项目。
关键词
二甲双胍
牛骨骼肌卫星细胞
细胞增殖
细胞分化
metformin
bovine skeletal muscle satellite cells
cell proliferation
cell differentiation
