不同加载时间流体剪切力对成骨细胞中piezo1机械敏感型蛋白表达的影响

Effects of loading time of fluid shear stress on expression of piezo1 mechanosensitive protein in osteoblasts

目的:研究加载不同时间流体剪切力(FSS)对MC3T3-E1成骨细胞piezo1机械敏感型离子蛋白表达的影响。方法:利用自行设计的平行平板FSS加载装置,对MC3T3-E1成骨细胞施加12 dyn/cm^2FSS 0、15、30、45、60、90 min,采用免疫荧光染色实验检测piezo1机械敏感型离子蛋白的表达水平。结果:piezo1明显表达于成骨细胞细胞质及细胞核,细胞质尤为明显。对体外培养的MC3T3-E1细胞加载12 dyn/cm^2FSS,随着加载时间的延长,piezo1蛋白表达上调,在45 min左右达到高峰。结论:加载不同时间的12 dyn/cm^2FSS能够上调MC3T3-E1成骨细胞piezo1机械敏感型离子蛋白,加载45 min最合适。

Objective To discuss the variations of the expression of piezo1 mechanosensitive ion protein in MC3T3-E1 osteoblasts with the increasing loading time of fluid shear stress（FSS）. Methods With the use of a self-designed parallel plate flow system,FSS（12 dyn/cm^2） was acted on MC3T3-E1 osteoblasts for 0, 15, 30, 45, 60, 90 min. The expression level of piezo1 mechanosensitive ion protein was detected by immunofluorescence. Results Piezo1 was obviously expressed in the cytoplasm and nucleus of osteoblasts, especially in the cytoplasm. FSS of 12 dyn/cm^2 was acted on MC3T3-E1 cells cultured in vitro. With the increase of loading time, the expression of piezo1 protein was upregulated. At 45 minutes after loading, the expression of piezo1 protein reached the maximum. Conclusion Loading 12 dyn/cm^2 FSS can upgrade the expression of piezo1 mechanosensitive ion protein in MC3 T3-E1 osteoblasts, and the optimal loading time of FSS is 45 minutes.