骨骼肌微损伤自体修复中超微结构变化与蛋白质组学研究

发布时间：2018-05-21 13:02

本文选题：微损伤 + 蛋白质组　；参考：《中国运动医学杂志》2015年03期

【摘要】：目的:从蛋白质组代谢角度分析骨骼肌微损伤自体修复过程中不同功能蛋白质组与肌纤维超微结构恢复的同步变化机制。方法:Wistar雄性大鼠36只随机分为安静对照组(D组)、运动后即刻组(0 h组)、运动后24 h组(24 h组)、运动后48 h组(48 h组)、运动后72 h组(72 h组)及运动后168 h组(168 h组)每组6只。运动方案:跑台下坡跑,坡度-16度,速度18 m/min,运动持续60 min,运动后相应时间处死。超薄切片、染色,用JEM-1200EX透射电镜分析不同组别骨骼肌的超微结构。以D组电泳图谱作参考,进行特异蛋白质表达量分析并对表达受到激活或者抑制的蛋白质进行进一步质谱鉴定。特异蛋白质按照功能不同分为I、II、III和IV类。结果:运动后即刻,参与细胞损伤修复的蛋白质(II类)包括膜联蛋白A5、维生素D结合蛋白、α1抗胰蛋白酶前体、DJ-1蛋白、免疫球蛋白、热休克蛋白β1、着丝粒关联蛋白E表达先于超微结构变化;运动后24~72 h,损伤修复过程中能量代谢相关蛋白质(I类)如辅酶Q9、烟酰胺腺嘌呤二核苷酸脱氢酶、乙酰基转移酶、异柠檬酸脱氢酶、磷酸甘油-3脱氢酶受到显著激活,可能促进了线粒体自身修复进而加快肌纤维再生速度;运动后48 h组和72 h组内Trim72蛋白、角蛋白、核糖体蛋白、肌球蛋白、载脂蛋白和内质网驻地蛋白29参与骨骼肌构成的结构蛋白质(III类)表达出现显著上升,这与超微结构恢复具有一定的同步性。结论:骨骼肌微损伤后0~48 h之内细胞损伤修复蛋白表达先于超微结构变化;在损伤修复后24~72 h之内线粒体酶系统及能量代谢相关蛋白表达先于超微结构修复,这可能促进了线粒体自身修复进而加快肌纤维再生速度;在损伤修复后24~48 h之内,骨骼肌结构蛋白表达与超微结构恢复具有一定的同步性。
[Abstract]:Aim: to analyze the synchronous changes of ultrastructure between different functional proteome and muscle fiber during autorepair of skeletal muscle microinjury from the perspective of proteome metabolism. Methods Thirty-six male Wistar rats were randomly divided into two groups: control group D (n = 36), group D (n = 6), group B (n = 6), group D (n = 6), group B (n = 24), group B (n = 48), group B (n = 48). Exercise plan: running downhill, slope-16 degrees, speed 18 m / min, exercise lasted for 60 minutes, the corresponding time after exercise death. Ultrathin sections were stained and ultrastructure of skeletal muscles of different groups were analyzed by JEM-1200EX transmission electron microscope. The specific protein expression was analyzed by using D group electrophoretic map as reference, and the expression of activated or inhibited protein was further identified by mass spectrometry. The specific proteins can be classified into III and IV according to their functions. Results: immediately after exercise, the proteins involved in the repair of cell injury included annexin A5, vitamin D binding protein, 伪 1 antitrypsin precursor DJ-1 protein, immunoglobulin. Heat shock protein 尾 1, centromere associated protein E expression prior to ultrastructural changes, energy metabolism related protein class I during injury repair at 24h after exercise, such as coenzyme Q9, nicotinamide adenine dinucleotide dehydrogenase, acetyltransferase, The activation of isocitrate dehydrogenase and glycerol phosphate dehydrogenase may promote the repair of mitochondria and accelerate the regeneration rate of muscle fiber, Trim72 protein, keratin, ribosomal protein, myosin in 48 h group and 72 h group after exercise. The expression of apolipoprotein and endoplasmic reticulum site protein 29 involved in structural protein class III) of skeletal muscle increased significantly, which was synchronized with the recovery of ultrastructure. Conclusion: the expression of cell injury repair protein was prior to ultrastructural changes within 48 hours after microinjury of skeletal muscle, and the expression of mitochondrial enzyme system and energy metabolism related protein was prior to ultrastructural repair within 24 to 72 hours after injury repair. This may promote the repair of mitochondria and accelerate the regeneration rate of muscle fiber, and the expression of skeletal muscle structural protein and the recovery of ultrastructure have a certain synchronism within 24 hours after repair.
【作者单位】：山东体育学院;中国科学院电工研究所生物医学工程部;泰山护理职业学院公共教学部;潍坊护理职业学院基础部;
【基金】：国家体育总局重点研究领域攻关课题(2014B059) 山东省自然科学基金(ZR2013CM034,ZR2013CL024) 山东省高校科技计划项目(J12LE04)
【分类号】：R873

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