P对大鼠骨骼肌卫星细胞化学损伤的保护作用
he Protective Effect of ATP on Skeletal Muscle Satellite Cells Damaged by H2O2
目的 研究ATP对大鼠骨骼肌卫星细胞H2O2损伤的保护作用及其对相关蛋白Bax和Bcl-2表达的影响，推进肌肉化学损伤保护与修复的研究。 方法 取新生大鼠后肢肌肉，组织块培养法进行体外培养，并通过传代培养，扩增卫星细胞，建立大鼠骨骼肌卫星细胞H2O2损伤模型。实验分成4组：正常对照组；模型对照组（0.1 mmol/L H2O2损伤4min）；损伤保护组（剂量为10、5、2.5、1.25、0.625、0.3125、0.15625 mg/mL ATP溶液分别处理细胞，然后0.1mmol/L H2O2损伤4min）；增殖对照组（剂量为10、5、2.5、1.25、0.625、0.3125、0.15625 mg/mL ATP溶液分别处理细胞）。并采用MTT法检测存活率，用Giemsa染色、FITC+PI+DAPI荧光凋亡检测、以及免疫细胞化学荧光抗体检测等方法研究损伤保护程度，后通过进行数据分析得出结论。 结果 大鼠骨骼肌卫星细胞经H2O2损伤后细胞存活率明显降低、凋亡率增加，ATP溶液能明显提高经H2O2损伤后细胞的存活率(P<0.01)，降低凋亡率；免疫荧光抗体检测表明ATP溶液能促进Bcl-2表达(P<0.01)，抑制Bax表达(P<0.01)；不同剂量ATP溶液对化学损伤的骨骼肌卫星细胞作用不同：添加适中浓度（2.5、1.25、0.625 mg/mL）ATP溶液的细胞存活率增大，浓度为1.25mg/mL ATP溶液的细胞存活率最大，加入较高浓度和较低浓度ATP溶液的细胞存活率降低，加入较高浓度ATP溶液的细胞甚至出现明显的凋亡。 结论 ATP对H2O2损伤的大鼠骨骼肌卫星细胞具有保护作用，其作用可能与通过JAK-STAT信号通路增强凋亡抑制基因Bcl-2的表达、抑制凋亡促进基因Bax的表达从而抑制细胞凋亡。
Objective To investigate the protective effect of ATP on skeletal muscle satellite cells of newborn rats damaged by H2O2 and its influence on the related proteins Bax and Bcl-2. Methods The H2O2-induced damage models of skeletal muscle satellite cells of newborn rats were established and the study was divided into 4 groups：normal control group，model control group (0.1 mmol / L H2O2 solution damaged cells for 50s)，protection group(10，5，2.5，1.25，0.625，0.3125，0.15625 mg/mL ATP solutions treated cells and then 0.1 mmol / L H2O2 solution damaged cells for 50s)，proliferation group (10，5，2.5，1.25，0.625，0.3125，0.15625 mg/mL ATP solutions treated cells). MTT colorimetric analysis，FITC+PI+DAPI fluorescent staining，Giemsa staining and immunocytochemistry fluorescent apoptotic antibody test were applied to detect the survival rate of cells and the protective effect of ATP on skeletal muscle satellite cells damaged by H2O2. Results The survival rate of skeletal muscle satellite cells was decreased and the apoptosis rate was increased after damaged. The survival rate of muscle satellite cells protected by the ATP solution could be increased (P<0.01), and the apoptosis rate was decreased. Immunocytochemistry staining analysis results showed that ATP solution could enhance Bcl-2 proteins (P<0.01) and inhibite Bax proteins (P<0.01). Different doses of ATP solutions had different effects on skeletal muscle satellite cells induced by H2O2: the survival rate of muscle satellite cells protected by the adequate doses of ATP solutions （2.5、1.25、0.625 mg/mL）could be increased. The protective effect on cells treated by 1.25 mg/mL ATP solution was the most obvious. Cells treated by 10，5mg/mL ATP solutions could even cause obvious apoptosis. Conclusion The ATP solution had obvious protective effect on skeletal muscle satellite cells damaged by H2O2 in new-born rats. The main mechanism of ATP solution was related to strengthen the expression of Bcl-2 and inhibit that of Bax through JAK -STAT signal transduction, thus inhibiting apoptosis of cells.
陶丽君、黄宝珠、张红红、费飞、朱道立
基础医学细胞生物学生理学
P过氧化氢损伤保护骨骼肌卫星细胞Bcl-2Bax
denosine-triphosphateHydrogen peroxideDamage and protectionSkeletal muscle satellite cellsBcl-2Bax
陶丽君,黄宝珠,张红红,费飞,朱道立.P对大鼠骨骼肌卫星细胞化学损伤的保护作用[EB/OL].(2013-04-27)[2025-08-10].http://www.paper.edu.cn/releasepaper/content/201304-550.点此复制
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