[1]沈欣欣,薛纯纯,杨佳凡,等.乌头原碱对小鼠成肌细胞增殖活性和成肌分化影响的实验研究[J].中医正骨,2025,37(10):1-6.
　SHEN Xinxin,XUE Chunchun,YANG Jiafan,et al.Effects of aconitine on the proliferative activity and myogenic differentiation of myoblasts in mice:an experimental study[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2025,37(10):1-6.
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乌头原碱对小鼠成肌细胞增殖活性和成肌分化影响的实验研究()

《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第37卷
期数:
2025年10期
页码:
1-6
栏目:
基础研究
出版日期:
2025-10-20

文章信息/Info

Title:
Effects of aconitine on the proliferative activity and myogenic differentiation of myoblasts in mice:an experimental study
作者:
沈欣欣1; 薛纯纯2; 杨佳凡2; 李煜楠2; 薛永鹏2; 杜文岚2; 李灵星2; 李晓锋2; 笪巍伟2
1.苏州市吴江区中医医院/苏州市吴江区第二人民医院,江苏 苏州 215221; 2.上海中医药大学附属市中医医院,上海 200071
Author(s):
SHEN Xinxin1; XUE Chunchun2; YANG Jiafan2; LI Yu'nan2; XUE Yongpeng2; DU Wenlan2; LI Lingxing2; LI Xiaofeng2; DA Weiwei2
1.Wujiang District Hospital of Traditional Chinese Medicine(The Second People's Hospital of Wujiang District),Suzhou 215221,Jiangsu,China 2.Shanghai Municipal Hospital of Traditional Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200071,China
关键词:
成肌细胞;  乌头碱;  肌; 骨骼;  细胞增殖;  成肌分化;  实验研究
Keywords:
myoblasts;  aconitine;  muscle; skeletal;  cell proliferation;  myoblast differentiation;  experimental study
摘要:
目的:观察乌头原碱对小鼠成肌细胞增殖活性和成肌分化的影响。方法:①乌头原碱对小鼠成肌细胞C2C12增殖活性影响的检测。将培养的小鼠成肌细胞C2C12分为5组(空白组、5 μmol·L-1组、10 μmol·L-1组、20 μmol·L-1组、40 μmol·L-1组),除空白组外,其他4组分别加入相应浓度的乌头原碱进行干预,计算各组细胞的增殖率,并筛选乌头原碱最佳干预浓度。②乌头原碱对小鼠成肌细胞C2C12成肌分化影响的检测。将培养的小鼠成肌细胞C2C12分为对照组和乌头原碱组。对照组加入马血清,乌头原碱组加入马血清和最佳干预浓度的乌头原碱进行干预。干预2 d和4 d后,分别采用逆转录PCR和Western Blot法检测2组小鼠成肌细胞C2C12中肌生成分化因子1(myogenic differentiation 1,MyoD1)、肌生成素(myogenin,MyoG)、肌球蛋白重链(myosin heavy chains,MyHC)的mRNA及蛋白表达水平; 并采用免疫荧光法检测2组小鼠成肌细胞C2C12中MyHC阳性表达情况,计算肌管融合指数。结果:①乌头原碱对小鼠成肌细胞C2C12增殖活性影响的检测结果。10 μmol·L-1、20 μmol·L-1、40 μmol·L-1组小鼠成肌细胞C2C12的细胞增殖率均高于空白组(P=0.000,P=0.000,P=0.000); 5 μmol·L-1组的细胞增殖率与空白组相比,差异无统计学意义(P=0.806); 20 μmol·L-1组的细胞增殖率高于10 μmol·L-1和40 μmol·L-1组(P=0.011,P=0.037)。②乌头原碱对小鼠成肌细胞C2C12成肌分化影响的检测结果。干预2 d和4 d后,乌头原碱组小鼠成肌细胞C2C12中MyoD1、MyoG、MyHC的mRNA和蛋白相对表达量及肌管融合指数均高于对照组(P值均>0.05)。结论:乌头原碱可增强小鼠成肌细胞的增殖活性,且可上调小鼠成肌细胞成肌分化相关基因的表达,促进其成肌分化。
Abstract:
Objective:To investigate the effects of aconitine on the proliferative activity and myogenic differentiation of myoblasts in mice.Methods:①Assessing the effect of aconitine on the proliferative activity of C2C12 myoblasts from mice.The cultured C2C12 myoblasts were divided into blank group,5 μmol/L group,10 μmol/L group,20 μmol/L group,and 40 μmol/L group.Except for the blank group,the C2C12 myoblasts in the other 4 groups were intervened with aconitine in their corresponding concentration,respectively,for consecutive 24 hours.After the end of intervention,the proliferation rate of C2C12 myoblasts in each group was calculated to identify the optimal intervention concentration of aconine.②Assessing the effect of aconitine on the myogenic differentiation of C2C12 myoblasts from mice.The cultured C2C12 myoblasts were divided into control group and aconitine group.The ones in the control group were intervened with horse serum,while the ones in the aconitine group with horse serum and aconitine at the predetermined optimal concentration.After 2- and 4-day intervention,the mRNA and protein expression levels of myogenic differentiation 1(MyoD1),myogenin(MyoG),and myosin heavy chains(MyHC)in the C2C12 myoblasts were detected by reverse transcription-PCR(RT-PCR)and Western Blot,respectively.Additionally,the MyHC-positive expression in the C2C12 myoblasts were detected by immunofluorescence assay,and the myotube fusion index was calculated.Results:①Effect of aconitine on the proliferative activity of C2C12 myoblasts.The proliferation rate of C2C12 myoblasts was higher in 10,20,and 40 μmol/L groups compared to blank group(P=0.000,P=0.000,P=0.000),with the 20 μmol/L group showing a superior effect over both 10 and 40 μmol/L groups(P=0.011,P=0.037),while,it showed no significant difference between 5 μmol/L group and blank group(P=0.806).②Effect of aconitine on the myogenic differentiation of C2C12 myoblasts.After 2- and 4-day intervention,the relative mRNA and protein expression levels of MyoD1,MyoG,and MyHC in the C2C12 myoblasts,as well as the myotube fusion index,were all higher in aconitine group compared to control group(all P-values were>0.05).Conclusion:Aconitine can enhance the proliferative activity of mouse myoblasts and upregulate the expression of myogenic differentiation-related genes,thereby promoting the myogenic differentiation.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金项目(82374471,82374473,81973881); 上海市东方英才计划青年项目(B02A2); 苏州市吴江区“科教兴卫”项目(WWK202325)
通讯作者:笪巍伟 E-mail:dww2348285@163.com
更新日期/Last Update: 1900-01-01