过氧化物酶体增殖物激活受体δ在原代小鼠成肌细胞分化过程中对肌球蛋白重链亚型的影响

荆海军; 周播江

doi:10.16098/j.issn.05290-1356.2020.05.021

PDF(10211 KB)

解剖学报 ›› 2020, Vol. 51 ›› Issue (5) : 765-771. DOI: 10.16098/j.issn.05290-1356.2020.05.021

细胞和分子生物学

过氧化物酶体增殖物激活受体δ在原代小鼠成肌细胞分化过程中对肌球蛋白重链亚型的影响

荆海军周播江*

作者信息 +

Effect of peroxisome proliferator-activated receptor δ on myosin heavy chain subtypes during the differentiation of primary mouse myoblasts

JING Hai-jun ZHOU Bo-jiang*

Author information +

文章历史 +

摘要

目的探讨过氧化物酶体增殖物激活受体δ(PPARδ)在成肌细胞分化过程中对肌球蛋白重链(MHC)亚型的影响。方法分离5只新生小鼠原代成肌细胞，设立对照组、PPARδ激动剂组和PPARδ拮抗剂组；各组细胞在分化培养基中培养2、4、6 d收取细胞样品，MHC免疫荧光鉴定各组细胞分化情况；Real-time PCR及免疫荧光检测MHC各亚型表达水平；Western blotting检测各组细胞MHC蛋白表达水平。结果成肌细胞分化过程中PPARδ激动剂组形成的肌管和MHC蛋白表达量高于对照组和PPARδ拮抗剂组。通过比较PPARδ激动剂处理后MHC 4个亚型随时间变化规律发现，分化2 d MHCⅠ、MHCⅡa 的上调作用显著；分化4 d MHCⅠ、MHCⅡa、MHCⅡx上调作用显著，其中MHCⅡa上调幅度最大；分化6 d MHCⅠ、MHCⅡa上调作用显著，其中MHCⅡa上调幅度最大。结论 PPARδ可以促进小鼠原代成肌细胞分化，在分化过程中显著上调MHCⅡa表达。

Abstract

Objective To study the effect of peroxisome proliferator-activated receptor δ (PPARδ) on myosin heavy chain (MHC) subtypes during myoblast differentiation. Methods Primary myoblasts from newborn mice were isolated, control group, PPAR delta agonist group and PPAR delta antagonist group were set up. Cell samples were collected at 2, 4 and 6 days of cell differentiation in each group. Identification of cell differentiation in each group was made by MHC immunofluorescence; The expression levels of MHC subtypes were detected by Real-time PCR; Western blotting was used to detect the MHC protein expression levels in each group. Results The expression of myotube and MHC protein in PPARδ agonist group were higher than that in control group and PPARδ antagonist group. By comparing the four MHC subtypes treated with PPARδ agonist, it was found that: after 2 days of differentiation, the up-regulation of MHCⅠ and MHCⅡa was significant; After 4 days of differentiation, the up-regulation of MHCⅠ, MHCⅡa and MHCⅡx was significant, and the up-regulation of MHCⅡa was the largest; After 6 days of differentiation, the up-regulation of MHCⅠ and MHCⅡa was significant, and the up-regulation of MHCⅡa was the largest. Conclusion PPARδ promotes mouse myoblast differentiation and significantly upregulates MHCⅡa expression during differentiation.

导出引用

荆海军周播江. 过氧化物酶体增殖物激活受体δ在原代小鼠成肌细胞分化过程中对肌球蛋白重链亚型的影响[J]. 解剖学报. 2020, 51(5): 765-771 https://doi.org/10.16098/j.issn.05290-1356.2020.05.021

JING Hai-jun ZHOU Bo-jiang. Effect of peroxisome proliferator-activated receptor δ on myosin heavy chain subtypes during the differentiation of primary mouse myoblasts[J]. Acta Anatomica Sinica. 2020, 51(5): 765-771 https://doi.org/10.16098/j.issn.05290-1356.2020.05.021

中图分类号： R322

参考文献

［1］ Slot IGM, Schols AMWJ, Vosse BAH, et al. Hypoxia differentially regulates muscle oxidative fiber type and metabolism in a HIF-1α-dependent manner［J］. Cell Signal, 2014, 26(9):1837-1845.
［2］ Koo JH, Kim TH, Park SY, et al. Gα13 ablation reprograms myofibers to oxidative phenotype and enhances whole-body metabolism［J］. J Clin Inv, 2017, 127(10):3845-3860.
［3］ Kr?mer DK, Ahlsén M, Norrbom J, et al. Human skeletal muscle fibre type variations correlate with PPARα, PPARδ and PGC-1αmRNA［J］. Acta Physiol, 2006, 188(3-4):207-216.
［4］ Zhu DL, Xu P, Xie F, et al. Function of histochemical type of skeletal muscle fiber in rat and myosin heavy chain［J］. Acta Anatomica Sinica, 2007, 38(1):93-97. (in Chinese)
朱道立, 徐萍, 谢锋, 等. 大鼠骨骼肌纤维组织化学分型与肌球蛋白重链的功能［J］. 解剖学报, 2007, 38(1):93-97.
［5］ Wang YM, Dai Y, Liu XF, et al. Isolation identification and induced differentiation of bovine sSkeletal muscle satellite cells［J］. Chinese Animal Husbandry and Veterinary Medicine, 2014, 41(7):142-147. (in Chinese)
王轶敏, 代阳, 刘新峰, 等. 牛骨骼肌卫星细胞的分离鉴定和诱导分化［J］. 中国畜牧兽医, 2014, 41(7):142-147.
［6］ Xiong L, He YJ, Dai W, et al. Isolation, purification and culture of primary mouse myoblast［J］. Journal of Modern Medicine & Health, 2017, 33(3):23-25. (in Chinese)
熊雷, 何衍佶, 戴威,等. 小鼠原代成肌细胞分离、纯化及培养［J］. 现代医药卫生, 2017, 33(3):23-25.
［7］ Matsuoka Y, Inoue A. Controlled differentiation of myoblast cells into fast and slow muscle fibers［J］. Cell Tissue Res, 2008, 332(1):123-132.
［8］ Wang M, Yu H, Kim YS, et al. Myostatin facilitates slow and inhibits fast myosin heavy chain expression during myogenic differentiation［J］. Biochem Biophys Res Commun, 2012, 426(1):83-88.
［9］ Zhu LN, Ren Y, Chen JQ, et al. Effects of myogenin on muscle fiber types and key metabolic enzymes in gene transfer mice and C2C12 myoblasts［J］. Gene, 2013, 532(2):246-252.
［10］ Zhang Y, Li W, Zhu M, et al. FHL3 differentially regulates the expression of MyHC isoforms through interactions with MyoD and pCREB［J］. Cell Signal, 2016, 28(1):60-73.
［11］ Choi YM, Chen PR, Shin S, et al. Mild heat stress enhances differentiation and proliferation of Japanese quail myoblasts and enhances slow muscle fiber characteristics［J］. Poult Sci, 2016, 95(8):1912-1917.