[1]刘众成,何良志,移穷,等.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,2022,39(10):1287-1292.[doi:DOI:10.3969/j.issn.1005-202X.2022.10.018]
　LIU Zhongcheng,HE Liangzhi,YI Qiong,et al.Piezo1 ion channel-mediated fluid shear stress inhibits the apoptosis of MLO-Y4 osteocytes[J].Chinese Journal of Medical Physics,2022,39(10):1287-1292.[doi:DOI:10.3969/j.issn.1005-202X.2022.10.018]
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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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《中国医学物理学杂志》[ISSN:1005-202X/CN:44-1351/R]

卷:
39卷
期数:
2022年第10期
页码:
1287-1292
栏目:
医学生物物理
出版日期:
2022-11-02

文章信息/Info

Title:
Piezo1 ion channel-mediated fluid shear stress inhibits the apoptosis of MLO-Y4 osteocytes
文章编号:
1005-202X（2022）10-1287-06
作者:
刘众成; 何良志; 移穷; 刘雪宁; 张坤; 王力夫; 耿彬; 夏亚一; 姜金
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
Author(s):
LIU Zhongcheng;  HE Liangzhi;  YI Qiong;  LIU Xuening;  ZHANG Kun;  WANG Lifu;  GENG Bin;  XIA Yayi;  JIANG Jin
Gansu Provincial Key Laboratory of Orthopaedics/Department of Orthopaedics, Lanzhou University Second Hospital, Lanzhou 730000, China
关键词:
流体剪切力; Piezo1; 骨细胞; 凋亡
Keywords:
Keywords: fluid shear stress Piezo1 osteocytes apoptosis
分类号:
R318；R329.2
DOI:
DOI:10.3969/j.issn.1005-202X.2022.10.018
文献标志码:
A
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。
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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备注/Memo

备注/Memo:
【收稿日期】2022-05-04 【基金项目】国家自然科学基金（81874017, 81960403, 82060405, 82160422）；甘肃省中医药管理局科研项目（TCM-IPC-2020-03）；兰州市科技计划项目（2021-RC-102，20JR5RA320）；兰州大学第二医院“萃英科技创新”计划（CY2017-ZD02，CY2021-MS-A07，CY2021-MS-B02） 【作者简介】刘众成，硕士，研究方向：机械应力与骨质疏松、运动医学，E-mail: liuzhch_14@163.com；何良志，硕士，研究方向：机械应力与骨质疏松、运动医学，E-mail: 976620652@qq.com（刘众成和何良志为共同第一作者） 【通信作者】姜金，博士，研究方向：细胞生物力学、运动医学，E-mail: jiangjin2007@163.com；夏亚一，博士，主任医师，教授，博士生导师，研究方向：关节外科、生物材料、细胞生物力学，E-mail: xiayylzu@126.com
更新日期/Last Update: 2022-10-27