Myoblast survival enhancement and transplantation success improvement by heat-shock treatment in MDX mice
Myoblast survival enhancement and transplantation success improvement by heat-shock treatment in MDX mice
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DOI：
10.1097/01.tp.0000121503.01535.f5
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发表时间：
2004-05-15
期刊：
影响因子：
6.2
通讯作者：
Tremblay, JP
中科院分区：
文献类型：
--
作者：
Bouchentouf, M;Benabdallah, BF;Tremblay, JP
背景。杜氏肌营养不良症是一种因无法合成肌营养不良蛋白而引起的疾病，肌营养不良蛋白与肌膜完整性的维持有关。成肌细胞移植是这种疾病的潜在治疗方法。然而，大多数移植细胞在注射后很快就会死亡。当细胞经历各种类型的应激时，热休克蛋白（HSP）会过度表达。因此，我们的目标是研究HSP（特别是HSP70）的表达是否可以保护成肌细胞在肌内注射后免于死亡。方法。通过将细胞在42℃下加热60分钟来诱导HSP70表达。通过蛋白质印迹分析对 HSP70 过表达进行定量。使用 Hoescht/碘化丙啶标记技术通过荧光激活细胞分选仪分析评估 HSP 对细胞存活的体外影响，并通过移植[甲基-C-14]胸苷标记的 TnI-LacZ 成肌细胞来研究其体内影响。结果。蛋白质印迹表明热休克处理后 HSP70 过表达七倍。在体外，热休克处理保护 18% 的细胞免受十字孢菌素（1 μM）诱导的细胞凋亡。 HSP 还保护 10% 的细胞免受肿瘤坏死因子-a (30 ng/mL) 或葡萄糖氧化酶 (0.1 U/mL) 诱导的死亡。在体内，该治疗使移植后 5 天的细胞存活率提高两倍，并使长期移植成功率提高四倍。结论。热休克治疗是提高成肌细胞移植成功率的实用方法；事实上，使用这种治疗方法，在移植前不需要对细胞进行基因改造。
Background. Duchenne muscular dystrophy is a disease caused by the incapacity to synthesize dystrophin, which is implicated in the maintenance of the sarcolemma integrity. Myoblast transplantation is a potential treatment of this disease. However, most of the transplanted cells die very rapidly after their injection. Heat-shock proteins (HSPs) are over-expressed when cells undergo various types of stresses. Our goal was thus to investigate whether the expression of HSPs (HSP70 in particular) could protect myoblasts from death after intramuscular injection.Methods. HSP70 expression was induced by warming the cells at 42degreesC for 60 minutes. HSP70 over-expression was quantified by Western blot analysis. The in vitro effect of HSPs on cell survival was evaluated by fluorescence-activated cell sorter analysis using the Hoescht/propidium iodide-labeling technique, and their in vivo effects were investigated by transplanting TnI-LacZ myoblasts labeled with [methyl-C-14] thymidine.Results. Western blots indicated a sevenfold overexpression of the HSP70 after the heat-shock treatment. In vitro, the heat-shock treatment protected 18% of the cells from staurosporine- (1 muM) induced apoptosis. HSPs also protected 10% of the cells from death induced by either tumor necrosis factor-a (30 ng/mL) or glucose oxydase (0.1 U/mL). In vivo, the treatment improved the cell survival by twofold 5 days after the graft and increased by fourfold the long-term graft success.Conclusions. The heat-shock treatment is a practical approach for improving the success of myoblast transplantation; in fact, using this kind of treatment, there is no need to genetically modify the cells before their transplantation.