Hidalgo Magdalena, Marchant Dominique, Quidu Patricia, Youcef-Ali Karima, Richalet Jean P, Beaudry Michele, Besse Sophie, Launay Thierry
Paris Cité Sorbonne, Université Paris 13, EA 2363 Réponses Cellulaires et Fonctionnelles à l'Hypoxie, Paris, France.
Cell Physiol Biochem. 2014;33(1):67-77. doi: 10.1159/000356650. Epub 2014 Jan 3.
This work aims to study the regulation of the glutathione peroxidase and catalase activities in myoblasts from the L6 line exposed to 21%, 5% and 1% O2 during the cell differentiation.
Rat L6 myoblasts were grown in 1%, 5% or 21% O2 in the presence or absence of N-acetyl cysteine. The cell proliferation was evaluated by determining the doubling time and kinetics of cultures by counting cells. The cell differentiation was analyzed by determining the myogenic fusion index using antibodies against the myosin heavy chain. The glutathione peroxidase and catalase activities were assayed. The p110-PI3K/Thr308-Akt pathway was studied using western blotting. The oxidative status of the cells was carried out by determining TBARS.
5% O2 improves the glutathione peroxidase activity, p110-PI3K/Thr308-Akt pathway and differentiation while 1% O2 alters all these parameters compared to 21% O2. NAC (0.5 mM) can prevent the deleterious effects of hypoxia (1% O2) on the L6 myoblast proliferation and enhances the myoblast differentiation when exposed to 21% O2. TBARS are reduced in 5% O2 compared to both 21% and 1% O2.
The glutathione peroxidase activity and p110-PI3K/Thr308-Akt are both modulated in the same way by oxygen.
本研究旨在探讨L6细胞系成肌细胞在细胞分化过程中暴露于21%、5%和1%氧气条件下时谷胱甘肽过氧化物酶和过氧化氢酶活性的调节情况。
大鼠L6成肌细胞在有或无N - 乙酰半胱氨酸存在的情况下,分别在1%、5%或21%氧气环境中培养。通过计数细胞来测定培养物的倍增时间和动力学,以此评估细胞增殖情况。使用抗肌球蛋白重链抗体测定成肌融合指数,分析细胞分化情况。检测谷胱甘肽过氧化物酶和过氧化氢酶活性。采用蛋白质免疫印迹法研究p110 - PI3K/Thr308 - Akt信号通路。通过测定丙二醛(TBARS)来评估细胞的氧化状态。
与21%氧气条件相比,5%氧气可提高谷胱甘肽过氧化物酶活性、激活p110 - PI3K/Thr308 - Akt信号通路并促进细胞分化,而1%氧气则改变了所有这些参数。N - 乙酰半胱氨酸(0.5 mM)可预防低氧(1%氧气)对L6成肌细胞增殖的有害影响,并且在细胞暴露于21%氧气时增强成肌细胞分化。与21%和1%氧气条件相比,5%氧气条件下丙二醛水平降低。
谷胱甘肽过氧化物酶活性和p110 - PI3K/Thr308 - Akt信号通路受氧气的调节方式相同。