过度机械拉伸应力通过Piezo1介导肌腱细胞凋亡的作用及机制研究

陈墨龙; 陈万; 唐康来

doi:10.16016/j.2097-0927.202302064

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过度机械拉伸应力通过Piezo1介导肌腱细胞凋亡的作用及机制研究

基础医学 | 更新时间：2026-01-09

- 过度机械拉伸应力通过Piezo1介导肌腱细胞凋亡的作用及机制研究
- Excessive mechanical stretch stress induces tendon cell apoptosis via mechanosensitive ion channel protein Piezo1
- 陆军军医大学学报 2023年45卷第10期页码：1040-1049
- 作者机构：
  
  400038 重庆，陆军军医大学(第三军医大学)第一附属医院运动医学中心
- 作者简介：
  
  [唐康来，E-mail：tangkanglai@hotmail.comTANG Kanglai, E-mail: tangkanglai@hotmail.com
- 基金信息：
- DOI：10.16016/j.2097-0927.202302064
  中图分类号： R322.74;R363.11;R363.21
- 收稿：2023-02-12，
  
  修回：2023-03-17，
  
  纸质出版：2023-05-30
- 稿件说明：
移动端阅览
陈墨龙, 陈万, 唐康来. 过度机械拉伸应力通过Piezo1介导肌腱细胞凋亡的作用及机制研究[J]. 陆军军医大学学报, 2023,45(10):1040-1049.

Molong CHEN, Wan CHEN, Kanglai TANG. Excessive mechanical stretch stress induces tendon cell apoptosis via mechanosensitive ion channel protein Piezo1[J]. Journal of Army Medical University, 2023, 45(10): 1040-1049.
陈墨龙, 陈万, 唐康来. 过度机械拉伸应力通过Piezo1介导肌腱细胞凋亡的作用及机制研究[J]. 陆军军医大学学报, 2023,45(10):1040-1049. DOI： 10.16016/j.2097-0927.202302064.

Molong CHEN, Wan CHEN, Kanglai TANG. Excessive mechanical stretch stress induces tendon cell apoptosis via mechanosensitive ion channel protein Piezo1[J]. Journal of Army Medical University, 2023, 45(10): 1040-1049. DOI： 10.16016/j.2097-0927.202302064.

摘要

目的

探索机械敏感离子通道蛋白Piezo1在过度机械拉伸应力诱导肌腱细胞凋亡中的作用机制。

方法

分离并培养8周龄雄性C57小鼠(

=20

体质量22~26 g)的肌腱细胞，采用Flexcell系统建立机械拉伸诱导肌腱细胞凋亡的细胞模型。将肌腱细胞分为对照组、20%拉伸组、20%拉伸+Yoda1组、20%拉伸+GsMTx4组、20%拉伸+Piezo1敲低慢病毒(Lv-Piezo1)组及20%拉伸+对照慢病毒(Lv-Ctrl)组，采用线粒体膜电位、流式细胞仪、Western blot和钙离子荧光探针检测Piezo1在肌腱细胞凋亡中的作用。将肌腱细胞分为20%拉伸组、20%拉伸+siRNA介导Calpain2敲低(si-Calpain2)组、20%拉伸+对照siRNA(si-Ctrl)组，Western blot检测Piezo1下游信号Calpain2/BAX/Caspase3轴的激活情况。

结果

过度机械拉伸应力作用下，20%拉伸组细胞凋亡水平显著高于对照组(

0.05)，Yoda1进一步促进了机械拉伸应力诱导的肌腱细胞凋亡(

0.05)，而GsMTx4和Lv-Piezo1表现出相反的作用(

0.05)。过度机械拉伸应力作用下，肌腱细胞Calpain2、BAX和cleaved-Caspase3的表达增强(

0.05)。20%拉伸+si-Calpain2组BAX和cleaved-Caspase3的表达水平低于20%拉伸组、20%拉伸+对照siRNA组(

0.05)，敲低Calpain2可抑制BAX和cleaved-Caspase3的表达(

0.05)，减轻肌腱细胞凋亡(

0.05)。

结论

过度机械牵拉应力通过激活Piezo1和下游Calpain2/BAX/Caspase3通路诱导肌腱细胞凋亡，Piezo1有望成为肌腱病的潜在治疗靶点。

Abstract

Objective

To investigate the role and mechanism of Piezo1

a mechanically-sensitive ion channel protein

in excessive mechanical stretch induced apoptosis of tendon cells.

Methods

Tendon cells of 8-week-old male C57 mice(

=20

body mass: 22~26 g) were isolated and cultured

and a cell model of mechanical stretch induced apoptosis of tendon cells was established using the Flexcell system. Tendon cells were divided into the control group

the 20% elongation group

the 20% elongation+Yoda1 group

the 20% elongation+GsMTx4 group

the 20% elongation+Piezo1 knockdown lentivirus gro

up (Lv-Piezo1) and 20% elongation+control lentivirus group (Lv-Ctrl). Mitochondrial membrane potential assay

flow cytometry assay

Western blot assay and calcium fluorescence probe were used to detect the role of Piezo1 in tendon cell apoptosis. Tendon cells were divided into the 20% elongation group

the 20% elongation+siRNA-mediated Calpain2 knockdown group (si-Calpain2)

and the 20% elongation+control siRNA group (si-Ctrl). Western blot assay was used to detect the activation of Piezo1 downstream signal Calpain2/BAX/Caspase3 axis.

Results

The level of apoptosis with excessive mechanical tensile stress was higher in the 20% elongation group than those in the control group (

0.05)

and Yoda1 promoted mechanical tensile stress induced apoptosis in tendon cells (

0.05)

while GsMTx4 and Lv-Piezo1 showed the opposite effect (

0.05). The expression of Calpain2

BAX and cleaved-Caspase3 in tendon cells was enhanced by excessive mechanical stretch stress (

0.05). The 20% elongation+si-Calpain2 group had lower expression levels of BAX and cleaved-Caspase3 than those in the 20% elongation and the 20% elongation+control siRNA group (

0.05). Knockdown of Calpain2 inhibited the expression of BAX and cleaved-Caspase3 (

0.05) and attenuated apoptosis in tendon cells (

0.05).

Conclusion

Excessive mechanical stretch stress can induce apoptosis of tendon cells

via

the activation of Piezo1 and the downstream Calpain2/BAX/Caspase3 pathways. Piezo1 holds promise as a potential therapeutic target for tendinopathy.

关键词

Keywords

references

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