碱性成纤维生长因子对大鼠成肌细胞氧化应激损伤的作用及其机制

Effects and mechanism of bFGF on rat myoblast oxidative injury induced by hydrogen peroxide

目的:探讨碱性成纤维生长因子(bFGF)下调氧自由基水平在大鼠成肌细胞氧化应激中的保护作用。方法:采用对数生长期大鼠成肌细胞,随机分为四组(n=6):正常对照组(control),单纯bFGF组(bFGF),氧化应激组(H_2O_2),干预组(bFGF+H_2O_2)。氧化应激组采用含100μmol/L H_2O_2的培养基继续孵育4 h。免疫组化检测B淋巴细胞瘤-2基因(Bcl-2)阳性沉积颗粒;免疫荧光观察活性氧自由基(ROS)水平、Bcl-2、B淋巴细胞瘤-2基因相关X蛋白(Bax)及细胞色素C(Cyt.C)表达;Western blot检测Cyt.C、聚腺苷二磷酸核糖聚合酶(PARP)蛋白表达情况。结果:与正常组比较,氧化应激组成肌细胞Bcl-2表达降低,ROS、Cyt.C表达显著增加(P<0.05)。与氧化应激组比较,bFGF干预后的成肌细胞Bcl-2表达增加,ROS及Cyt.C表达减少(P<0.05)。结论:bFGF对成肌细胞氧化应激具有保护作用,其机制可能与bFGF下调氧自由基水平、Bcl-2蛋白增加、Cyt.C表达减少有关。

Objective： To explore the protective role of basic fibreblast growth factor （bFGF） on attenuating hydrogen peroxide-induced injury in cultured rat myoblasts. Methods： Cultured rat myoblasts at growth phase were randomly divided into four groups （ n = 6） ： control group （control）, bFGF group （bFGF）, model group（H202） and the treatment group （bFGF ＋ H2O2）. Model group was treated with 100 μmol/L hydrogen peroxide for 4h. B-cell lymphoma-2 （Bcl-2） positive particles were detected by immunohistochemistry; Reactive oxygen species （ROS） and expression for Bcl-2 associated X protein （Bax）, Bcl-2 and Cytochrome C （Cyt. C） fluorescence were observed under the invented microscope; Cyt. C and Poly ADP-ribose polymerase（PARP）protein were assessed by Western blot. Results： Compared with control group, the myoblats in the model group showed low expression of Bcl-2 positive particles, accompanied by high expression of ROS level and Cyt. C fuorescence （ P 〈 0.05） ; Compared with medel group, bFGF enhanced Bcl-2 activity of the myoblasts, and significantly downregulated Cyt. C and PARP expression （ P 〈 0.05）. Conclusion： bFGF could attenuate oxidative injury of rat myoblasts induced by hydrogen peroxide, which mechanism might be related to enhanced Bcl-2 and reduced ROS, Cyt. C levels.