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[1]倪钰鸽,盛 菲,牛文彦.环氧化酶2抑制剂parecoxib对C2C12骨骼肌细胞分化的影响及机制研究[J].天津医科大学学报,2024,30(02):138-143.[doi:10.20135/j.issn.1006-8147.2024.02.0138]
　NI Yuge,SHENG Fei,NIU Wenyan.Study of effect and mechanism of cyclooxygenase-2 inhibitor parecoxib on the differentiation of C2C12 skeletal muscle cells[J].Journal of Tianjin Medical University,2024,30(02):138-143.[doi:10.20135/j.issn.1006-8147.2024.02.0138]
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环氧化酶2抑制剂parecoxib对C2C12骨骼肌细胞分化的影响及机制研究(PDF)
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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:
30卷
期数:
2024年02期
页码:
138-143
栏目:
基础医学
出版日期:
2024-03-20

文章信息/Info

Title:
Study of effect and mechanism of cyclooxygenase-2 inhibitor parecoxib on the differentiation of C2C12 skeletal muscle cells
文章编号:
1006-8147（2024）02-0138-06
作者:
倪钰鸽; 盛 菲; 牛文彦
（天津医科大学基础医学院免疫学系，天津300070）
Author(s):
NI Yuge; SHENG Fei; NIU Wenyan
（Department of Immunology，School of Basic Medical Sciences，Tianjin Medical University，Tianjin 300070，China）
关键词:
环氧化酶2抑制剂; 骨骼肌; 肌原纤维; 细胞分化
Keywords:
cyclooxygenase-2 inhibitors;  skeletal muscle;  myofibril;  cell differentiation
分类号:
R392.1
DOI:
10.20135/j.issn.1006-8147.2024.02.0138
文献标志码:
A
摘要:
目的：探讨环氧化酶2抑制剂帕瑞昔布（parecoxib）对C2C12小鼠骨骼肌细胞分化的影响及其分子机制。方法：CCK-8法检测不同浓度的parecoxib处理后C2C12细胞的活力；分化培养基诱导C2C12小鼠骨骼肌细胞48 h后，将细胞分为对照组（control组）和parecoxib组继续分化48 h。免疫荧光检测肌管细胞分化成熟的标志蛋白肌球蛋白重链（MyHC），qPCR和Western印迹分别检测MyHC、肌原纤维Ⅰ型（Myh7）、Ⅱa型（Myh2）、Ⅱb型（Myh4）、Ⅱx型（Myh1）、肌生成决定因子（MyoD）、肌生成因子5（Myf5）、肌生成素（myogenin）的基因和蛋白表达。结果：0、100、150、200、250、300 ?滋mol/L的parecoxib均不影响细胞活力。与对照组相比，parecoxib组MyHC阳性肌纤维数量减少，肌管融合受损，分化程度降低，MyHC、MyHCⅡa、MyHCⅡb、MyHCⅡx、MyoG、MyoD和Myf5的mRNA水平降低（t=19.04、53.93、72.38、33.72、15.32、3.061、18，均P＜0.05），MyHC Ⅰ的mRNA水平升高（t=17.12，P＜0.01）。Western印迹结果显示，与对照组相比，parecoxib组MyHC、MyHCⅡa、MyHCⅡb、MyHCⅡx、MyoG和MyoD蛋白水平降低（t=7.297、7.852、11.43、227、80.14、11.76，均P＜0.01），MyHC Ⅰ蛋白水平升高（t=5.891，P＜0.01）。结论：Parecoxib可能通过下调MyoG、MyoD和Myf5的表达，抑制C2C12骨骼肌细胞的分化。
Abstract:
Objective：To investigate the effect of parecoxib，a cyclooxygenase-2 inhibitor，on the differentiation of C2C12 skeletal muscle cells and its potential molecular mechanism. Methods：The Cell Counting Kit-8（CCK-8） was used to detect the viability of C2C12 cells treated with different concentrations of parecoxib. C2C12 mouse skeletal muscle cells were incubated with differentiation medium for 48 hours. Then the cells were divided into control group and parecoxib group for further differentiation for 48 hours. Immunofluorescence was used to detect myosin heavy chain（MyHC），a marker of differentiation and maturation of myotubule cells. The mRNA and protein expression of MyHC，MyHCI（Myh7），Ⅱa（Myh2），Ⅱb（Myh4），Ⅱx（Myh1），myogenic determinant（MyoD），myogenic factor 5（Myf5） and Myogenin were detected by qPCR and Western blotting，respectively. Results：0，100，150，200，250 and 300 ?滋mol/L parecoxib did not affect cell viability. Immunofluorescence data showed that compared with the control group，the number of MyHC-positive muscle fibers in the parecoxib group was reduced，muscle tube fusion was impaired，and the degree of differentiation was reduced. The results of qPCR showed that compared with the control group，the mRNA levels of MyHC，MyHCⅡa，MyHCⅡb，MyHCⅡx，MyoG，MyoD and Myf5 in the parecoxib group were decreased（t=19.04，53.93，72.38，33.72，5.32，3.061，18，all P<0.05），and the mRNA level of MyHC Ⅰ was increased（t=17.12，P<0.01）. Western blotting results showed that compared with the control group，the protein levels of MyHC，MyHCⅡa，MyHCⅡb，MyHCⅡx，MyoG and MyoD in the parecoxib group were decreased（t=7.297，7.852，11.43，227， 80.14， 11.76，all P<0.01），and the MYHC Ⅰ protein level was increased（t= 5.891，P<0.01）. Conclusion：Parecoxib may inhibit the differentiation of C2C12 skeletal muscle cells by down-regulating the expression of MyoG，MyoD and Myf5.

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备注/Memo

备注/Memo:
基金项目 国家自然科学基金面上项目（82270856）
作者简介 倪钰鸽（1998-），女，硕士在读，研究方向：医学检验技术；通信作者：牛文彦，E-mail：wniu@tmu.edu.cn。
更新日期/Last Update: 2024-03-20