Steiner Jennifer L, Pruznak Anne M, Deiter Gina, Navaratnarajah Maithili, Kutzler Lydia, Kimball Scot R, Lang Charles H
Department of Cellular and Molecular Physiology, and Surgery, Penn State College of Medicine, Hershey, Pennsylvania, United States of America.
PLoS One. 2014 Jun 19;9(6):e99582. doi: 10.1371/journal.pone.0099582. eCollection 2014.
Sepsis decreases skeletal muscle protein synthesis in part by impairing mTOR activity and the subsequent phosphorylation of 4E-BP1 and S6K1 thereby controlling translation initiation; however, the relative importance of changes in these two downstream substrates is unknown. The role of 4E-BP1 (and -BP2) in regulating muscle protein synthesis was assessed in wild-type (WT) and 4E-BP1/BP2 double knockout (DKO) male mice under basal conditions and in response to sepsis. At 12 months of age, body weight, lean body mass and energy expenditure did not differ between WT and DKO mice. Moreover, in vivo rates of protein synthesis in gastrocnemius, heart and liver did not differ between DKO and WT mice. Sepsis decreased skeletal muscle protein synthesis and S6K1 phosphorylation in WT and DKO male mice to a similar extent. Sepsis only decreased 4E-BP1 phosphorylation in WT mice as no 4E-BP1/BP2 protein was detected in muscle from DKO mice. Sepsis decreased the binding of eIF4G to eIF4E in WT mice; however, eIF4E•eIF4G binding was not altered in DKO mice under either basal or septic conditions. A comparable sepsis-induced increase in eIF4B phosphorylation was seen in both WT and DKO mice. eEF2 phosphorylation was similarly increased in muscle from WT septic mice and both control and septic DKO mice, compared to WT control values. The sepsis-induced increase in muscle MuRF1 and atrogin-1 (markers of proteolysis) as well as TNFα and IL-6 (inflammatory cytokines) mRNA was greater in DKO than WT mice. The sepsis-induced decrease in myocardial and hepatic protein synthesis did not differ between WT and DKO mice. These data suggest overall basal protein balance and synthesis is maintained in muscle of mice lacking both 4E-BP1/BP2 and that sepsis-induced changes in mTOR signaling may be mediated by a down-stream mechanism independent of 4E-BP1 phosphorylation and eIF4E•eIF4G binding.
脓毒症部分通过损害mTOR活性以及随后4E-BP1和S6K1的磷酸化来降低骨骼肌蛋白质合成,从而控制翻译起始;然而,这两种下游底物变化的相对重要性尚不清楚。在基础条件下以及对脓毒症的反应中,评估了野生型(WT)和4E-BP1/BP2双敲除(DKO)雄性小鼠中4E-BP1(和-BP2)在调节肌肉蛋白质合成中的作用。12月龄时,WT和DKO小鼠的体重、瘦体重和能量消耗没有差异。此外,DKO和WT小鼠腓肠肌、心脏和肝脏的体内蛋白质合成速率没有差异。脓毒症使WT和DKO雄性小鼠的骨骼肌蛋白质合成和S6K1磷酸化程度相似地降低。脓毒症仅降低了WT小鼠中4E-BP1的磷酸化,因为在DKO小鼠的肌肉中未检测到4E-BP1/BP2蛋白。脓毒症降低了WT小鼠中eIF4G与eIF4E的结合;然而,在基础或脓毒症条件下,DKO小鼠中eIF4E•eIF4G的结合没有改变。WT和DKO小鼠中均观察到脓毒症诱导的eIF4B磷酸化有类似增加。与WT对照值相比,WT脓毒症小鼠以及对照和脓毒症DKO小鼠肌肉中的eEF2磷酸化同样增加。脓毒症诱导的肌肉中MuRF1和atrogin-1(蛋白水解标志物)以及TNFα和IL-6(炎性细胞因子)mRNA的增加在DKO小鼠中比WT小鼠中更大。脓毒症诱导的心肌和肝脏蛋白质合成的降低在WT和DKO小鼠之间没有差异。这些数据表明,在缺乏4E-BP1/BP2的小鼠肌肉中,总体基础蛋白质平衡和合成得以维持,并且脓毒症诱导的mTOR信号变化可能由独立于4E-BP1磷酸化和eIF4E•eIF4G结合的下游机制介导。