地鳖肽提取物对H_(2)O_(2)刺激C2C12细胞肌调节因子基因表达的影响  

Effects of Eupofyphaga sinensis peptide extracts on expression of myogenic regulatory factor in C2C12 cells stimulated with H_(2)O_(2)

作　　者：陈靖阳 徐小艾 王婉 黄山 沈红 CHEN Jingyang;XU Xiaoai;WANG Wan;HUANG Shan;SHEN Hong(Animal Science and Technology College,Beijing University of Agriculture,Beijing 102206,China;Dental Medical Devices Testing Center,Peking University Hospital of Stomatology,Beijing 100081,China;Beijing Key Laboratory of Traditional Chinese Veterinary Medicine,Beijing University of Agriculture,Beijing102206,China)

机构地区：[1]北京农学院动物科学技术学院/动物类国家级实验教学示范中心,北京102206 [2]北京大学口腔医学院口腔医疗器械检验中心,北京100081 [3]北京农学院兽医学(中医药)北京市重点实验室,北京102206

出　　处：《中国兽医学报》2022年第3期551-557,共7页Chinese Journal of Veterinary Science

基　　金：北京市教委科研计划一般资助项目(KM201910020006);动物源食品安全教学团队建设资助项目(BUA2017JG068)。

摘　　要：探究地鳖肽提取物(Eupofyphaga sinensis peptide extract,ESP)对过氧化氢(H_(2)O_(2))刺激的C2C12细胞肌调节因子基因表达的影响。体外培养C2C12细胞其中培养基分别添加ESP和叔丁基对苯二酚(TBHQ)处理细胞诱导其分化融合9 d,除对照组外所有组细胞在试验结束前8 h培养基添加H_(2)O_(2)处理细胞;采用RT-qPCR和免疫荧光法检测肌调节因子基因和蛋白表达,细胞染色和显微镜观察细胞形态。对照组可见诱导分化融合9 d的C2C12细胞出现成熟多核肌管,随着细胞培养时间延长肌调节因子MyoG基因表达增加;与对照组比较,H_(2)O_(2)组C2C12细胞增殖调节因子(Pax7、Myf5、MyoD)基因表达显著降低,在细胞分化不同时间C2C12细胞分化调节因子(MyoD、MyoG、Myf6)mRNA表达显著下调,表达MyoG细胞阳性率和肌管融合率比对照组明显降低;与H_(2)O_(2)组比较,ESP组C2C12细胞增殖调节因子(Pax7、Myf5、MyoD)和细胞分化调节因子(MyoD、MyoG、Myf6)基因表达水平显著增加,表达MyoG细胞阳性率明显增加,其中细胞染色与镜下观察结果趋势一致。ESP能影响氧化应激C2C12细胞肌调节因子(Pax7、Myf5、MyoD、MyoG、Myf6)的基因表达,提高细胞增殖和分化融合能力,其机制可能与PGC-1α的激活有关。The experiment aims to investigate the effect of Eupofyphaga sinensis peptide(ESP)extracts on the expression of myogenic regulatory factor gene in C2C12cells stimulated with hydrogen peroxide(H_(2)O_(2)).C2C12cells were cultured in vitroin the medium containing ESP or TB-HQ to induce C2C12cells to differentiate and fuse for 9d,and H_(2)O_(2)was added to the medium for8hbefore the end of the experiment in all groups except the control group.RT-qPCR and immunofluorescence methods were used to detect the gene and protein expression of myogenic regulatory factor,and cells were stained by Giemsa and their morphology were observed by microscopy.In the control group,mature multinuclear myotubes were observed in C2C12cells which were induced to differentiate and fuse for 9d.Compared with the control group,the expression of Pax7,Myf5,MyoD gene of cell proliferation regulator in C2C12cells in H_(2)O_(2)group decreased significantly,and the expression of MyoD,MyoG,Myf6mRNA of cell differentiation regulator decreased obviously at different time of cell differentiation.The positive rate of cells expressing MyoG and myotube fusion rate in H_(2)O_(2)group were significantly lower than those in the control group.Compared with H_(2)O_(2)group,the expression level of Pax7,Myf5,MyoD gene of cell proliferation regulator and MyoG,Myf6gene of cell differentiation regulator in C2C12cells in ESP groups was significantly increased,and the positive rate of cells expressing MyoG was remarkably increased.The trend of cell staining was consistent with that of microscopic observation.In conclusion,ESP can affect the expression of muscle regulatory factors(Pax7,Myf5,MyoD,MyoG,Myf6)in C2C12cells under oxidative stress,and improve cell proliferation,differentiation and fusion by a mechanism that may be related to the activation of PGC-1α.

关 键 词：地鳖肽提取物 C2C12细胞 增殖及分化融合 肌调节因子 

分 类 号：S853.74[农业科学—临床兽医学]