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目的研究随机回转(RPM)模拟失重条件对微管微丝交联因子1(MACF1)表达及β-catenin信号通路的影响,为失重性骨丢失机制研究提供实验依据。方法 RPM条件培养成骨细胞,PCR检测MACF1mRNA表达变化,Western blot检测MACF1以及不同磷酸化位点β-catenin蛋白表达变化,荧光素酶报告基因检测细胞β-catenin荧光素酶活性变化。结果 RPM模拟失重条件下,成骨细胞MACF1表达下降,β-catenin mRNA和不同位点磷酸化蛋白表达量发生改变;MACF1 shRNA抑制了成骨细胞中β-catenin表达,影响了β-catenin蛋白稳定性及活性。结论 RPM模拟失重抑制了MACF1表达和β-catenin表达及活性;MACF1 shRNA影响了β-catenin表达及活性。MACF1可能是一种力敏感分子,通过调控β-catenin参与成骨细胞对力学信号的感知和响应。
Objective To investigate the effects of simulated weightlessness (RPM) on the expression of microtubule cross-linking factor 1 (MACF1) and β-catenin signaling in microtubules and provide experimental evidence for the mechanism of loss-of-mass bone loss. Methods Osteoblasts were cultured with RPM. The expression of MACF1 mRNA was detected by PCR. The expression of MACF1 and β-catenin at different phosphorylation sites were detected by Western blot. The changes of β-catenin luciferase activity were detected by luciferase reporter assay. Results Under simulated RPM conditions, the expression of MACF1 decreased and the expression of β-catenin mRNA and phosphorylated protein at different sites changed. MACF1 shRNA inhibited the expression of β-catenin in osteoblasts and affected the stability of β-catenin protein Sex and activity. Conclusion RPM simulated weight loss inhibited the expression of MACF1 and β-catenin, and the activity of MMP-1 shRNA influenced the expression of β-catenin. MACF1 may be a force-sensitive molecule that participates in the perception and response of mechanical signals to osteoblasts by regulating β-catenin.