Endoplasmic Reticulum Stress Markers and Ubiquitin-Proteasome Pathway Activity in Response to a 200-km Run
Endoplasmic Reticulum Stress Markers and Ubiquitin-Proteasome Pathway Activity in Response to a 200-km Run
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DOI：
10.1249/mss.0b013e3181e4c5d1
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发表时间：
2011-01-01
期刊：
影响因子：
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通讯作者：
Francaux, Marc
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作者：
Kim, Hyo Jeong;Jamart, Cecile;Francaux, Marc
金，h. j ., c. jamart, l. deldicque, g .。李彦宏，金志强，金志明。和弗朗克斯先生。内质网应激标志物和泛素-蛋白酶体途径活性对200公里跑步的响应。医学科学。体育Exerc。，第43卷第1期，第18-25页，2011年。目的：本研究调查了200公里跑步是否会调节骨骼肌细胞应激相关的信号通路，特别关注内质网应激和泛素-蛋白酶体通路的激活。方法：8名男子跑200 km （28 h 03 min + 2 h 01 min）。在赛前2周和赛后3小时分别对股外侧肌进行了两次肌肉活检。采用Western blot或实时定量聚合酶链反应检测丝裂原活化蛋白激酶、泛素-蛋白酶体途径、内质网应激、炎症和氧化应激标志物。用荧光法测定蛋白酶体的凝乳胰蛋白酶样活性。结果：胞外信号相关激酶1/2 （+401% +/- 173.8%,P = 0.027）和c-Jun n端（+149% +/- 61.9%,P = 0.023）磷酸化水平在比赛后升高，而p38磷酸化水平保持不变。BiP升高（+235% +/- 94.7%,P = 0.021），总信使RNA升高（+138% +/- 31.2%,P = 0.002），剪接的X-box结合蛋白1升高（+241% +/- 53.3%,P = 0.001）表明内质网应激的存在。炎症标志物白介素-6 （+403% +/- 96.1%,P = 0.002）、肿瘤坏死因子- α (+233% +/- 58.4%, P = 0.003)以及氧化应激标志物金属硫蛋白1F （+519% +/- 258.3%, P = 0.042）、金属硫蛋白1H （+666% +/- 157.5%, P = 0.002）、烟酰胺腺嘌呤二核苷酸磷酸氧化酶（NADPH氧化酶）（+162% +/- 60.5%,P = 0.016）的转录均升高。泛素连接酶肌肉特异性RING finger 1 （+583% +/- 244.3%, P = 0.024）、肌萎缩F-box （+249% +/- 83.8%, P = 0.011）和C2蛋白酶体亚基（+116% +/- 40.6%,P = 0.012）的信使RNA水平也有所升高。令人惊讶的是，泛素结合蛋白的数量和蛋白酶体的凝糜蛋白酶样活性分别下降了20% +/- 8.3% （P = 0.025）和21% +/- 4.4% （P = 0.001）。泛素特异性蛋白酶28去泛素酶表达升高（+81% +/- 37.9%,P = 0.034）。结论：在骨骼肌中，200公里跑步激活了泛素连接酶肌肉特异性RING finger 1和肌肉萎缩F-box的表达，以及各种细胞应激，其中包括内质网应激、氧化应激和炎症。同时，代偿机制似乎也被触发：未折叠蛋白反应被上调，蛋白酶体的凝乳胰蛋白酶样活性被抑制。
KIM, H. J., C. JAMART, L. DELDICQUE, G.-L. AN, Y. H. LEE, C. K. KIM, J.-M. RAYMACKERS, and M. FRANCAUX. Endoplasmic Reticulum Stress Markers and Ubiquitin-Proteasome Pathway Activity in Response to a 200-km Run. Med. Sci. Sports Exerc., Vol. 43, No. 1, pp. 18-25, 2011. Purpose: This study investigated whether a 200-km run modulates signaling pathways implicated in cellular stress in skeletal muscle, with special attention paid to the endoplasmic reticulum (ER) stress and to the activation of the ubiquitin-proteasome pathway. Methods: Eight men ran 200 km (28 h 03 min + 2 h 01 min). Two muscle biopsies were obtained from the vastus lateralis muscle 2 wk before and 3 h after the race. Mitogen-activated protein kinase, ubiquitin-proteasome pathway, ER stress, inflammation, and oxidative stress markers were assayed by Western blot analysis or by quantitative real-time polymerase chain reaction. Chymotrypsin-like activity of the proteasome was measured by a fluorimetric assay. Results: Phosphorylation states of extracellular signal-related kinase 1/2 (+401% +/- 173.8%, P = 0.027) and c-Jun N-terminal (+149% +/- 61.9%, P = 0.023) increased after the race, whereas p38 phosphorylation remained unchanged. Increases in BiP (+235% +/- 94.7%, P = 0.021) and in the messenger RNA level of total (+138% +/- 31.2%, P = 0.002) and spliced X-box binding protein 1 (+241% +/- 53.3%, P = 0.001) indicated the presence of ER stress. Transcripts of inflammatory markers interleukin-6 (+403% +/- 96.1%, P = 0.002) and tumor necrosis factor-alpha (+233% +/- 58.4%, P = 0.003) as well as oxidative stress markers metallothionein 1F (+519% +/- 258.3%, P = 0.042), metallothionein 1H (+666% +/- 157.5%, P = 0.002), and nicotinamide adenine dinucleotide phosphate-oxidase (NADPH oxidase) (+162% +/- 60.5%, P = 0.016) were increased. The messenger RNA level of the ubiquitin ligases muscle-specific RING finger 1 (+583% +/- 244.3%, P = 0.024) and muscle atrophy F-box (+249% +/- 83.8%, P = 0.011) and the C2 proteasome subunit (+116% +/- 40.6%, P = 0.012) also increased. Surprisingly, the amount of ubiquitin-conjugated proteins and the chymotrypsin-like activity of the proteasome were decreased by 20% +/- 8.3% (P = 0.025) and 21% +/- 4.4% (P = 0.001), respectively. The expression of ubiquitin-specific protease 28 deubiquitinase was increased (+81% +/- 37.9%, P = 0.034). Conclusions: In the skeletal muscle, a 200-km run activates the expression of ubiquitin ligases muscle-specific RING finger 1 and muscle atrophy F-box as well as various cellular stresses, among which are ER stress, oxidative stress, and inflammation. Meanwhile, compensatory mechanisms seem also triggered: the unfolded protein response is up-regulated, and the chymotrypsin-like activity of the proteasome is repressed.