【摘要】 低温环境是东北寒区动物难以规避的重要应激源，可导致机体全身代谢紊乱与蛋白质合成重塑。RBM3可在低温刺激下快速上调表达，进而拮抗冷应激引发的损伤效应，因此被认定为一种重要的冷休克蛋白。研究旨在阐明急性冷应激条件下RBM3对断奶仔猪骨骼肌葡萄糖代谢的调控作用及细胞促存活效应，并验证OGT介导RBM3上调的分子机制。研究将9头“杜×长×大”三元杂交断奶仔猪随机分为常温对照组、冷刺激4 h组及冷刺激8 h组。待4℃急性冷刺激处理结束后，采集试验仔猪血液样本与骨骼肌组织。采用ELISA法测定各组仔猪血清CORT、CRH、E、NE、血糖及骨骼肌组织中FDP、PA含量；采用Western blot(WB)检测骨骼肌组织中OGT、RBM3、AKT、GLUT4、PFKFB2、GSK3β的蛋白表达及其磷酸化水平；采用高效液相色谱法测定骨骼肌组织中ATP、ADP、AMP含量。结果表明，急性冷应激可使断奶仔猪血清CORT、CRH、E、NE水平显著升高、血糖含量降低；骨骼肌组织中OGT、RBM3、AKT、GLUT4、PFKFB2、GSK3β蛋白表达及磷酸化水平均上调，FDP、PA含量升高，而ATP、ADP、AMP含量组间无显著差异。综上，急性冷刺激可诱发断奶仔猪产生明显冷应激反应；RBM3上调可促进AKT磷酸化激活，进而调控骨骼肌葡萄糖代谢重编程并抑制细胞凋亡，OGT或为介导RBM3快速上调的关键调控靶点。更多还原

【Abstract】 Low temperature is an inevitable major stressor for animals in cold regions of Northeast China,which can cause systemic metabolic disorders and protein synthesis remodeling in the body.As an important cold shock protein,RBM3 is rapidly upregulated upon low-temperature stimulation and further antagonizes the damage induced by cold stress.This study aimed to elucidate the regulatory role of RBM3 in skeletal muscle glucose metabolism and its pro-survival effect in weaned piglets under acute cold stress,and to verify the molecular mechanism by which OGT mediats the upregulation of RBM3.Nine “Duroc×Landrace×Yorkshire” crossbred weaned piglets were randomly divided into normal temperature control group,4 h cold stimulation group and 8 h cold stimulation group.After acute cold stress treatment at 4 ℃,blood samples and skeletal muscle tissues were collected.The serum levels of CORT,CRH,E,NE and blood glucose,as well as the contents of FDP and PA in skeletal muscle were measured by ELISA.Western blotting(WB)was used to detect the protein expression and phosphorylation levels of OGT,RBM3,AKT,GLUT4,PFKFB2,and GSK3β in skeletal muscle.The contents of ATP,ADP,and AMP in skeletal muscle were measured by high-performance liquid chromatography.The results showed that acute cold stress significantly increased serum CORT,CRH,E and NE levels while decreased blood glucose concentration in weaned piglets.The protein expression and phosphorylation levels of OGT,RBM3,AKT,GLUT4,PFKFB2,and GSK3β,as well as the contents of FDP and PA were upregulated in skeletal muscle,whereas no significant differences were observed in ATP,ADP,and AMP contents among groups.In conclusion,acute cold stimulation induces obvious cold stress responses in weaned piglets.The upregulation of RBM3 promotes the phosphorylation and activation of AKT,thereby regulating skeletal muscle glucose metabolic reprogramming and inhibiting cell apoptosis.OGT may serve as a key regulatory target mediating the rapid upregulation of RBM3.更多还原