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目的:探讨Wnt/β-catenin信号转导通路在H9c2心肌细胞慢性缺氧适应中的作用及可能机制。方法:将37℃、5%O_2、5%CO_2、90%N_2条件下培养的H9c2细胞作为实验组,低氧培养6、12、24、36、48、72 h时收集细胞;将37℃、21%O_2、5%CO_2条件下培养的细胞作为对照组。采用Western blot和real-time PCR技术检测β-catenin、抗凋亡因子bcl-2和survivin在不同培养时间点的表达变化。结果:5%O_2培养的H9c2细胞与对照组相比,β-catenin蛋白在缺氧6、12、24、36 h时表达明显增加(P=0.000,P=0.018,P=0.001,P=0.000),bcl-2及survivin表达较对照组明显增加(P=0.000,P=0.000)。结论:Wnt/β-catenin信号通路激活可能是H9c2心肌细胞慢性缺氧的适应机制之一,其部分可能通过上调抗凋亡因子bcl-2及survivin表达起作用。
Objective: To investigate the role of Wnt / β-catenin signal transduction pathway in H9c2 cardiomyocyte chronic hypoxia adaptation and its possible mechanism. Methods: H9c2 cells cultured in 37 ℃, 5% O 2, 5% CO 2 and 90% N 2 were used as experimental group. Cells were harvested at 6, 12, 24, 36, 21% O2, 5% CO2 under the conditions of cells cultured as a control group. The expression of β-catenin, anti-apoptotic factors bcl-2 and survivin at different culture time points were detected by Western blot and real-time PCR. Results: Compared with the control group, the expression of β-catenin in H9c2 cells cultured in 5% O_2 increased significantly at 6, 12, 24 and 36 h (P = 0.000, P = 0.001, P = ), the expression of bcl-2 and survivin increased significantly compared with the control group (P = 0.000, P = 0.000). Conclusion: Activation of Wnt / β-catenin signaling pathway may be one of the mechanisms of H9c2 cardiomyocyte chronic hypoxia, which may partly by up-regulating the expression of anti-apoptotic factors bcl-2 and survivin.