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  • Protein translation, proteolysis and autophagy in human skeletal muscle atrophy after spinal cord injury.

Protein translation, proteolysis and autophagy in human skeletal muscle atrophy after spinal cord injury.

Acta physiologica (Oxford, England) (2018-02-10)
L S Lundell, M Savikj, E Kostovski, P O Iversen, J R Zierath, A Krook, A V Chibalin, U Widegren
摘要

Spinal cord injury-induced loss of skeletal muscle mass does not progress linearly. In humans, peak muscle loss occurs during the first 6 weeks postinjury, and gradually continues thereafter. The aim of this study was to delineate the regulatory events underlying skeletal muscle atrophy during the first year following spinal cord injury. Key translational, autophagic and proteolytic proteins were analysed by immunoblotting of human vastus lateralis muscle obtained 1, 3 and 12 months following spinal cord injury. Age-matched able-bodied control subjects were also studied. Several downstream targets of Akt signalling decreased after spinal cord injury in skeletal muscle, without changes in resting Akt Ser473 and Akt Thr308 phosphorylation or total Akt protein. Abundance of mTOR protein and mTOR Ser2448 phosphorylation, as well as FOXO1 Ser256 phosphorylation and FOXO3 protein, decreased in response to spinal cord injury, coincident with attenuated protein abundance of E3 ubiquitin ligases, MuRF1 and MAFbx. S6 protein and Ser235/236 phosphorylation, as well as 4E-BP1 Thr37/46 phosphorylation, increased transiently after spinal cord injury, indicating higher levels of protein translation early after injury. Protein abundance of LC3-I and LC3-II decreased 3 months postinjury as compared with 1 month postinjury, but not compared to able-bodied control subjects, indicating lower levels of autophagy. Proteins regulating proteasomal degradation were stably increased in response to spinal cord injury. Together, these data provide indirect evidence suggesting that protein translation and autophagy transiently increase, while whole proteolysis remains stably higher in skeletal muscle within the first year after spinal cord injury.

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Sigma-Aldrich
丽春红S染液(零售包装) 溶液, BioReagent, suitable for electrophoresis, 0.1 % (w/v) in 5% acetic acid
Sigma-Aldrich
OxyBlot 蛋白质氧化检测试剂盒, The OxyBlot Protein Oxidation Detection Kit provides the reagents to perform the immunoblot detection of carbonyl groups introduced into proteins by oxidative reactions with ozone or oxides of nitrogen or by metal catalyzed oxidation.
Sigma-Aldrich
抗p62/SQSTM1 兔抗, ~1 mg/mL, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
抗LC3 兔抗, ~1 mg/mL, affinity isolated antibody, buffered aqueous solution
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蛋白酶抑制剂混合物套装I- Calbiochem, A cocktail of five protease inhibitors that will inhibit a broad range of proteases and esterases. Supplied with a data sheet.