Men Xiao-Ming, Xu Zi-Wei, Tao Xin, Deng Bo, Qi Ke-Ke
Institute of Animal Husbandry and Veterinary Science, Zhejiang Academy of Agricultural Science, Hangzhou, Zhejiang, China.
PeerJ. 2021 Apr 19;9:e11065. doi: 10.7717/peerj.11065. eCollection 2021.
Irisin (a glycosylated protein) is cleaved from fibronectin type III domain-containing protein 5 (FNDC5), which is expressed mainly in animal muscle tissues and has multiple metabolic regulatory activities. However, their roles in controlling myofiber types in skeletal muscle remain unclear.
Two different commercial hybridized pigs, LJH (a crossed pig containing Chinese native pig genotypes) and DLY (Duroc × Landrace × Yorkshire) were selected to analyze mRNA expression and the mRNA composition of four adult myosin heavy chain () isoforms (IIIaIIxIIb) in the (LD) muscle. C2C12 myoblasts were cultured to investigate the effects of on the four MyHCs mRNA expressive levels, using small interfering RNA for depletion and a eukaryotic expression vector carrying for overexpression. ZLN005 (a small molecule activator of FNDC5's upstream control gene ) or recombinant human irisin protein were also used.
In LD muscle, LJH pigs had the higher mRNA level, and proportion than DLY pigs (). For C2C12 cells in vitro, small interfering RNA (si-592) silencing of expression markedly reduced mRNA levels (), while overexpression significantly increased mRNA levels (). Exogenous irisin increased the mRNA levels of (peroxisome proliferator-activated receptor gamma coactivator 1-alpha), , , , (nuclear respiratory factor 1), (vascular endothelial growth factor), and (mitochondrial transcription factor A,) (), and the enzyme activities of SDH (succinate dehydrogenase), CK (creatine kinase), and MDH (malate dehydrogenase) in C2C12 myotubes (). These results showed that mRNA expression had a significant association with the characteristics of myofiber types in porcine muscle, and participated in regulating mRNA expression of C2C12 myogenic differentiation cells in vitro. could be an important factor to control muscle fiber types, which provides a new direction to investigate pork quality via muscle fiber characteristics.
鸢尾素(一种糖基化蛋白)由含III型纤连蛋白结构域蛋白5(FNDC5)裂解产生,FNDC5主要在动物肌肉组织中表达,具有多种代谢调节活性。然而,它们在控制骨骼肌肌纤维类型中的作用仍不清楚。
选择两种不同的商业杂交猪,LJH(一种含有中国本土猪基因型的杂交猪)和DLY(杜洛克×长白×大白),分析背最长肌(LD)中四种成人肌球蛋白重链(MyHC)亚型(IIIa、IIx、IIb)的mRNA表达和mRNA组成。培养C2C12成肌细胞,使用小干扰RNA进行敲低和携带FNDC5的真核表达载体进行过表达,以研究FNDC5对四种MyHCs mRNA表达水平的影响。还使用了ZLN005(FNDC5上游控制基因的小分子激活剂)或重组人鸢尾素蛋白。
在LD肌肉中,LJH猪的MyHC mRNA水平和IIx比例高于DLY猪(P<0.05)。对于体外培养的C2C12细胞,小干扰RNA(si-592)沉默FNDC5表达显著降低了MyHCs mRNA水平(P<0.05),而FNDC5过表达显著增加了MyHCs mRNA水平(P<0.05)。外源性鸢尾素增加了C2C12肌管中PPARGC1A(过氧化物酶体增殖物激活受体γ共激活因子1α)、PGC1B、TFAM、NRF1(核呼吸因子1)、VEGF(血管内皮生长因子)和MTCO1(线粒体转录因子A)的mRNA水平(P<0.05),以及琥珀酸脱氢酶(SDH)、肌酸激酶(CK)和苹果酸脱氢酶(MDH)的酶活性(P<0.05)。这些结果表明,FNDC5 mRNA表达与猪肌肉中肌纤维类型的特征显著相关,并参与调节体外C2C12成肌分化细胞的MyHCs mRNA表达。FNDC5可能是控制肌肉纤维类型的重要因素,这为通过肌肉纤维特征研究猪肉品质提供了新方向。