Effects of heat stress and mechanical stretch on protein expression in cultured skeletal muscle cells
Effects of heat stress and mechanical stretch on protein expression in cultured skeletal muscle cells
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DOI：
10.1007/s00424-003-1177-x
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发表时间：
2003-11-01
影响因子：
4.5
通讯作者：
Yoshioka, T
中科院分区：
文献类型：
--
作者：
Goto, K;Okuyama, R;Yoshioka, T
在培养体系中研究了热应激、机械拉伸或两者联合对热休克蛋白（HSPs）表达和总蛋白水平的影响。大鼠骨骼肌细胞（L 6）在软底培养板上培养。他们受到以下四种情况之一的考验：（1）在37 ℃孵育97小时，（2）在41 ℃孵育1小时，然后在37 ℃孵育96小时，（3）在37 ℃孵育1小时，然后循环拉伸96小时（初始长度的18%，2秒拉伸和4秒释放）或（4）在41 ℃下孵育1小时，然后在37 ℃下循环拉伸96小时。在粗匀浆、上清液和沉淀中测定HSP 72和HSP 90以及总蛋白的表达。通过热应力和/或机械应力（拉伸）增加匀浆和沉淀中的细胞蛋白浓度。注意到加热和拉伸的组合对匀浆和颗粒中蛋白质浓度的累积效应。热应激和/或牵张均能增加颗粒中HSP 72和HSP 90的表达。然而，热应激和/或拉伸后，上清液中的HSP 90没有变化。骨骼肌细胞中HSP 72和HSP 90表达的调节可能与总蛋白密切相关，其丰度也受到机械和热应激的刺激。这些观察结果强烈表明，加热和被动拉伸肌肉可能是有用的，作为增加肌肉质量的一种手段，不仅在运动员，而且在康复期间的患者。
Effects of heat stress, mechanical stretching or a combination of both on the expression of heat shock proteins (HSPs) and total protein level were studied in a culture system. Rat skeletal muscle cells (L6) were cultured on flexible-bottomed culture plates. They were subjected to one of the four following conditions: (1) 97 h incubation at 37 degreesC, (2) 1 h incubation at 41 degreesC followed by 96 h incubation at 37 degreesC, (3) 1 h incubation at 37 degreesC followed by 96 h cyclic stretching (18% of initial length, 2-s stretch and 4-s release) at 37 degreesC or (4) 1 h incubation at 41 degreesC followed by 96 h cyclic stretching at 37 degreesC. The expression of HSP72 and HSP90 and total protein was determined in the crude homogenates, supernatant and pellets. Cellular protein concentrations in the homogenates and pellets were increased by heat stress and/or mechanical stress (stretch). A cumulative effect of the combination of heating and stretch on the protein concentration in the homogenates and in the pellets was noted. The expressions of HSP72 and HSP90 in the pellets were also increased by heat stress and/or stretch. However, HSP90 in the supernatant did not change following heat stress and/or stretch. The regulation of HSP72 and HSP90 expression in skeletal muscle cells may be closely related to total protein, the abundance of which is also stimulated by mechanical and heat stresses. These observations suggest strongly that heating and passive stretch of muscle may be useful as a means of increasing muscle mass, not only in athletes but also in patients during rehabilitation.