AMP-Activated Protein Kinase Abundance and Activation is increased with the β-Adrenergic agonist Zilpaterol Hydrochloride in Muscle from Feedlot Cattle

B. Johnson
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Abstract

The objective of this research was to evaluate the effect of Zilpaterol Hydrochloride (ZH) on myogenic or adipogenic gene and protein expression in skeletal muscle. Two feeding trials and one cell culture experiments were conducted. Semimembranosus muscle tissue was collected from steers that had been fed a diet containing 8.3 mg of ZH/kg DM for the last 0 or 20 d of the finishing period with a 3 d withdrawal period. To test the mode of action an in vitromodel was used with, isolated bovine satellite cells isolated from muscle tissue. Real Time-QPCR (RTQPCR) was used to measure the relative mRNA abundance of Adenosine Monophosphate Protein Kinase α (AMPK), Myosin Heavy Chain (MHC) I, MHCIIA, MHCIIX, Insulin-like Growth Factor I (IGF-I), β-adrenergic receptor (βAR) 1 and 2, peroxisome proliferator-activated receptor gamma (PPARγ), and Stearoyl-CoA Desaturase (SCD). Western blotting was used to measure the relative protein abundance of AMPK and Phosphorylated-AMPK (pAMPK). No differences were detected in relative mRNA abundance of AMPK, MHCIIA, IGF-I, βAR1 and βAR2. However, MHCI, SCD, and PPARγ mRNA expression was decreased and MHCIIX mRNA increased from ZH fed cattle compared to non-ZH. For one experiment, AMPK protein expression increased, while in another experiment, AMPK phosphorylation increased with ZH fed animals. The increase in MHCIIX mRNA with ZH fed cattle indicated the start of a fiber type shift towards larger diameter fibers. This shift may have been due to increased expression and phosphorylation of AMPK. These data suggest that the shift increase in MHCIIX was likely due to the ZH administration.
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β-肾上腺素能激动剂盐酸Zilpaterol可提高饲料牛肌肉中amp活化蛋白激酶的丰度和活性
本研究旨在探讨盐酸Zilpaterol (ZH)对骨骼肌中成肌或成脂基因及蛋白表达的影响。进行了2个饲养试验和1个细胞培养试验。在育肥期的最后0或20 d饲喂8.3 mg ZH/kg DM的饲粮,并进行3 d的停饲期,收集半膜肌组织。为了检验其作用方式,我们用牛肌肉组织分离的卫星细胞建立了体外模型。Real Time-QPCR (RTQPCR)检测各组细胞中腺苷单磷酸蛋白激酶α (AMPK)、肌球蛋白重链(MHC) I、MHCIIA、MHCIIX、胰岛素样生长因子I (IGF-I)、β-肾上腺素能受体(βAR) 1和2、过氧化物酶体增殖物激活受体γ (PPARγ)和硬脂酰辅酶a去饱和酶(SCD) mRNA的相对丰度。Western blotting检测AMPK和磷酸化AMPK (pAMPK)的相对蛋白丰度。AMPK、MHCIIA、IGF-I、βAR1和βAR2 mRNA相对丰度无差异。然而,与非ZH相比,ZH饲牛MHCI、SCD和PPARγ mRNA表达降低,MHCIIX mRNA表达升高。在一个实验中,AMPK蛋白表达增加,而在另一个实验中,AMPK磷酸化随着ZH喂养动物而增加。ZH饲牛MHCIIX mRNA的增加表明纤维类型开始向大直径纤维转移。这种转变可能是由于AMPK的表达和磷酸化增加。这些数据表明MHCIIX的转移增加可能是由于ZH的施用。
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