| Piezo1离子通道介导的流体剪切应力抑制MLO-Y4骨细胞凋亡 |
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| 引用本文： | 刘众成,何良志,移穷,刘雪宁,张坤,王力夫,耿彬,夏亚一,姜金.Piezo1离子通道介导的流体剪切应力抑制MLO-Y4骨细胞凋亡[J].中国医学物理学杂志,2022,0(10):1287-1292. |
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| 作者姓名： | 刘众成 何良志 移穷 刘雪宁 张坤 王力夫 耿彬 夏亚一 姜金 |
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| 作者单位： | 兰州大学第二医院骨科/甘肃省骨与关节疾病重点实验室， 甘肃 兰州 730000 |
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| 摘 要： | 目的:研究Piezo1机械敏感性离子通道对骨细胞的调控作用。方法:应用课题组研发的流体剪切力（FSS）加载装置，使用免疫荧光探究FSS对Piezo1通道在MLO-Y4中影响的最佳强度；对Piezo1使用FSS、激动剂Yoda1或阻滞剂GsMTx4进行干预。使用Western Blot探究Cleaved Caspase-3、Bcl-2、Bax凋亡相关蛋白的表达量；使用Hoechst-33258以及流式细胞凋亡检测MLO-Y4骨细胞的凋亡比例。结果:免疫荧光染色结果表明FSS对MLO-Y4骨细胞中Piezo1的表达呈现先增加后降低的单峰趋势，其中以9 dyne/cm2加载30 min为最佳刺激条件。使用Yoda1抑制MLO-Y4细胞凋亡（P<0.05），使用GsMTx4促进MLO-Y4细胞凋亡（P<0.05）。Piezo1介导的FSS抑制MLO-Y4细胞凋亡（P<0.05）。结论:Piezo1机械敏感性离子通道介导的FSS在9 dyne/cm2条件下抑制MLO-Y4骨细胞凋亡，且FSS在该条件下可以有效逆转Piezo1阻滞剂GsMTx4在4 μmol/L作用0.5 h下促进MLO-Y4细胞凋亡作用。
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| 关 键 词： | 流体剪切力 Piezo1 骨细胞 凋亡 |
Piezo1 ion channel-mediated fluid shear stress inhibits the apoptosis of MLO-Y4 osteocytes |
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| LIU Zhongcheng,HE Liangzhi,YI Qiong,LIU Xuening,ZHANG Kun,WANG Lifu,GENG Bin,XIA Yayi,JIANG Jin.Piezo1 ion channel-mediated fluid shear stress inhibits the apoptosis of MLO-Y4 osteocytes[J].Chinese Journal of Medical Physics,2022,0(10):1287-1292. |
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| Authors: | LIU Zhongcheng HE Liangzhi YI Qiong LIU Xuening ZHANG Kun WANG Lifu GENG Bin XIA Yayi JIANG Jin |
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| Affiliation: | Gansu Provincial Key Laboratory of Orthopaedics/Department of Orthopaedics, Lanzhou University Second Hospital, Lanzhou 730000, China |
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| Abstract: | Abstract: Objective To investigate the regulation of mechanosensitive ion channel Piezo1 on osteocytes. Methods Immunofluorescence staining was used to investigate the best condition for Piezo1 channel-mediated FSS loaded by the self-developed fluid shear stress (FSS) loading device on MLO-Y4 osteocytes. Piezo1 was intervened with FSS, Piezo1 activator Yoda1, or Piezo1 blocker GsMTx4. The expression levels of apoptosis-related proteins (Cleaved Caspase-3, Bcl-2, and Bax) were analyzed using Western Blot analysis, and the apoptosis rates of MLO-Y4 osteocytes was detected by Hoechst-33258 and flow cytometry. Results The result of immunofluorescence staining indicated that in MLO-Y4 osteocytes exposed to FSS, the expression of Piezo1 had a unimodal tendency of increasing firstly and then decreasing, and that the optimal stimulation condition was 9 dyne/cm2 loading for 30 minutes. Yoda1 inhibited the apoptosis of MLO-Y4 osteocytes (P<0.05), and GsMTx4 promoted the apoptosis of MLO-Y4 osteocytes (P<0.05). Piezo1-mediated FSS inhibited the apoptosis of MLO-Y4 osteocytes (P<0.05). Conclusion When loading FSS at 9 dyne/cm2, the FSS mediated by mechanosensitive ion channel Piezo1 can inhibit the apoptosis of MLO-Y4 osteocytes, and FSS can effectively reverse the apoptosis of MLO-Y4 osteocytes promoted with Piezo1 blocker GsMTx4 at 4 μmol/L for 0.5 h. |
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| Keywords: | Keywords: fluid shear stress Piezo1 osteocytes apoptosis |
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