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目的观察在不同应激条件下成肌细胞C2C12中Id2的表达与细胞分布情况,并探讨其表达与分布变化的机制。方法体外培养C2C12细胞,分别采用不同浓度H2O2氧化应激诱导细胞增殖或凋亡,不同浓度的促炎症信号LPS诱导细胞增殖,以及2%马血清诱导细胞分化。利用RT-PCR比较不同应激条件下Id2 mRNA的表达,同时利用免疫荧光检测Id2蛋白的表达强度和细胞分布。结果25μmol/l和50μmol/l的H_2O_2诱导下,Id2 mRNA表达比对照组分别增高80.5%和55.5%,荧光显著增强,Id2呈现细胞核与细胞质均匀分布。100 ng/ml和500 ng/ml的LPS亦诱导Id2 mRNA表达,较对照组增高21.7%和40.2%,荧光增强,但此时Id2以细胞核分布为主,细胞质少量分布。在2%马血清诱导成肌细胞分化后,Id2 mRNA表达显著降低,荧光显著减弱,并以细胞质分布为主。结论不同应激条件下Id2在成肌细胞中呈现不同的表达和分布,与其对骨骼肌损伤后再生的调控作用密切相关,表明Id2是骨骼肌损伤后再生的重要调控分子。
Objective To observe the expression and distribution of Id2 in C2C12 myoblasts under different stress conditions and to explore the mechanism of its expression and distribution changes. Methods C2C12 cells were cultured in vitro. Oxidative stress of H2O2 was used to induce cell proliferation or apoptosis. Different concentrations of proinflammatory signal LPS induced cell proliferation, and 2% horse serum induced cell differentiation. The expression of Id2 mRNA under different stress conditions was compared by RT-PCR, and the expression intensity and cell distribution of Id2 protein were detected by immunofluorescence. Results The expression of Id2 mRNA increased by 80.5% and 55.5%, respectively, under the induction of 25μmol / L and 50μmol / L H 2 O 2. The fluorescence intensity of Id2 mRNA increased significantly. Id2 showed uniform distribution of nucleus and cytoplasm. The expression of Id2 mRNA was also induced by 100 ng / ml and 500 ng / ml LPS, which was increased by 21.7% and 40.2% compared with the control group, and the fluorescence was enhanced. However, Id2 mainly distributed in nucleus and little cytoplasm. After 2% horse serum induced myoblast differentiation, Id2 mRNA expression was significantly reduced, the fluorescence decreased significantly, and mainly cytoplasmic distribution. Conclusion Id2 shows different expression and distribution in myoblasts under different stress conditions, which is closely related to its regulation on the regeneration of skeletal muscle after injury, indicating that Id2 is an important regulator of regeneration after skeletal muscle injury.