过度机械应力调控Piezo1介导成软骨细胞铁死亡的机制探讨

doi:10.11958/20241259

天津医药 ›› 2025, Vol. 53 ›› Issue (1): 14-18.doi: 10.11958/20241259

过度机械应力调控Piezo1介导成软骨细胞铁死亡的机制探讨

吴斌¹(), 刘兆祥², 张月红³, 王昌耀⁴^,^△()

1 青岛大学临沂市人民医院骨科（邮编276000）
2 沂水县第二人民医院骨科
3 东明县人民医院骨二科
4 青岛大学医学部附属医院关节外科

收稿日期:2024-09-03 修回日期:2024-11-03 出版日期:2025-01-15 发布日期:2025-02-06
通讯作者: ^△E-mail:swxg.1@163.com
作者简介:吴斌（1982），男，主治医师，主要从事创伤骨科方面研究。E-mail:jackay_wu@163.com
基金资助:
国家自然科学基金资助项目(81772329)

The mechanism of excessive mechanical stress modulates Piezo1-mediated ferroptosis in chondrocytes

WU Bin¹(), LIU Zhaoxiang², ZHANG Yuehong³, WANG Changyao⁴^,^△()

1 Department of Orthopedics, Qingdao University Linyi People’s Hospital, Linyi 276000, China
2 Department of Orthopedics, Yishui Second People’s Hospital
3 Department of Second Orthopedics, Dongming People’s Hospital
4 Department of Joint Surgery, the Affiliated Hospital of Qingdao University Medical College

Received:2024-09-03 Revised:2024-11-03 Published:2025-01-15 Online:2025-02-06
Contact: ^△E-mail:swxg.1@163.com

吴斌, 刘兆祥, 张月红, 王昌耀. 过度机械应力调控Piezo1介导成软骨细胞铁死亡的机制探讨[J]. 天津医药, 2025, 53(1): 14-18.

WU Bin, LIU Zhaoxiang, ZHANG Yuehong, WANG Changyao. The mechanism of excessive mechanical stress modulates Piezo1-mediated ferroptosis in chondrocytes[J]. Tianjin Medical Journal, 2025, 53(1): 14-18.

摘要/Abstract

摘要：

目的探讨过度机械应力调控Piezo1通道诱发小鼠成软骨细胞铁死亡的作用机制。方法选取小鼠ATDC5成软骨细胞系进行实验，使用siRNA-Piezo1干扰质粒和Piezo1过表达质粒转染细胞，给予机械应力刺激（MS），构建对照组（Control组）、MS组、MS+siRNA-Piezo1组（MS+sh组）和MS+Piezo1过表达组（MS+OV组）。用细胞增殖和毒性检测法（CCK-8）检测各组细胞增殖活力，透射电镜观察线粒体的形态和结构，生化检测氧化应激和亚铁离子（Fe²⁺）水平，流式细胞术检测脂质活性氧（ROS）水平，蛋白免疫印迹法（Western blot）检测溶质载体家族7成员11（SLC7A11）、谷胱甘肽过氧化物酶4（GPX4）、Ⅱ型胶原蛋白（Collagen Ⅱ）、基质金属蛋白酶（MMP）-13、蛋白聚糖（Aggrecan）及p53蛋白的表达水平。结果与Control组比较，MS组细胞活力下降，Fe²⁺、ROS、丙二醛（MDA）水平升高，还原型谷胱甘肽（GSH）、超氧化物歧化酶（SOD）水平降低（均P<0.05），电镜示线粒体嵴减少，SLC7A11、GPX4、Collagen Ⅱ、Aggrecan蛋白水平下降，p53和MMP-13蛋白水平则升高（均P<0.05）。与MS组比较，MS+sh组的Fe²⁺、ROS、MDA水平下降，GSH、SOD水平升高（均P<0.05），SLC7A11、GPX4、Collagen Ⅱ、Aggrecan蛋白水平升高，MMP-13和p53蛋白水平下降（均P<0.05）。与MS组比较，MS+OV组的细胞活力下降，Fe²⁺、ROS、MDA水平升高，GSH、SOD水平降低，SLC7A11、GPX4、Collagen Ⅱ、Aggrecan蛋白水平下降，MMP-13和p53蛋白水平上升（均P<0.05）。结论过度机械应力能够通过Piezo1通道蛋白介导成软骨细胞铁死亡，促进细胞外基质降解。

关键词: 骨关节炎, 软骨细胞, 应力, 物理, 铁死亡, Piezo1

Abstract:

Objective To explore the mechanism of excessive mechanical stress regulated ferroptosis induced by Piezo1 channel in mouse chondrocytes.Methods The experiment was performed on mouse ATDC5 chondrocytes. siRNA-Piezo1 interference plasmid and Piezo1 overexpression plasmid were used to transfect chondrocytes, and mechanical stress stimulation was given. The control group, the mechanical stress stimulation group (MS group), the MS+siRNA-Piezo1 group (MS+sh group) and the MS+Piezo1 overexpression group (MS+OV group) were constructed, respectively. The cell viability, Fe²⁺, ROS levels, the expression of ferroptosis-related proteins SLC7A11 and GPX4, and the expression of Collagen Ⅱ, MMP-13, Aggrecan and p53 proteins were detected in each group.Results Compared with the control group, the cell viability was decreased in the MS group (P<0.05). Levels of Fe²⁺, reactive oxygen species (ROS) and malondialdehyde (MDA) were increased (P<0.05). Levels of reduced glutathione (GSH) and superoxide dismutase (SOD) were decreased (P<0.05), and the mitochondrial ridge was decreased detected by transmission electron microscopy. Protein levels of SLC7A11, GPX4, Collagen Ⅱ and Aggrecan were decreased (P<0.05), while protein levels of p53 and MMP-13 were increased (P<0.05). Compared with the MS group, Fe²⁺, ROS and MDA levels were decreased in the MS+sh group (P<0.05), GSH and SOD levels were increased (P<0.05), and protein levels of SLC7A11, Collagen Ⅱ, GPX4 and Aggrecan were increased (P<0.05). The protein levels of MMP-13 and p53 were decreased (P<0.05). Compared with the MS group, cell viability was decreased (P<0.05), Fe²⁺, ROS and MDA levels were increased (P<0.05), GSH and SOD levels were decreased (P<0.05), and protein levels of SLC7A11, Collagen Ⅱ, GPX4 and Aggrecan were decreased in the MS+OV group (P<0.05). Levels of MMP-13 and p53 protein were increased (P<0.05).Conclusion Excessive mechanical stress can induce chondrocyte ferroptosis and promote extracellular matrix degradation via Piezo1 channel protein.

Key words: osteoarthritis, chondrocytes, stress, mechanical, ferroptosis, Piezo1

中图分类号:

R684.3

图/表 5

参考文献 24

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组别	24 h	48 h	72 h	F
Control组	100.00±4.96	120.54±5.07^A	131.24±3.88^AB	106.312^＊＊
MS组	70.75±5.11^a	74.64±5.26^aA	80.94±6.42^aAB	1.665^＊＊
MS+sh组	83.13±4.37^ab	94.47±4.83^abA	99.40±7.85^abAB	56.011^＊＊
MS+OV组	61.24±5.77^ab	62.49±5.45^ab	66.32±5.80^abAB	2.461^＊
F	31.933^＊＊	28.122^＊＊	30.851^＊＊

组别	24 h	48 h	72 h	F
Control组	100.00±4.96	120.54±5.07^A	131.24±3.88^AB	106.312^＊＊
MS组	70.75±5.11^a	74.64±5.26^aA	80.94±6.42^aAB	1.665^＊＊
MS+sh组	83.13±4.37^ab	94.47±4.83^abA	99.40±7.85^abAB	56.011^＊＊
MS+OV组	61.24±5.77^ab	62.49±5.45^ab	66.32±5.80^abAB	2.461^＊
F	31.933^＊＊	28.122^＊＊	30.851^＊＊

组别	Fe²⁺/（μmol/kg）		GSH/（mmol/g）		SOD/（U/mg）
组别		ROS/%		MDA/（μmol/g）
Control组		2.46±0.22		3.15±0.71
MS组		25.76±2.63^a		8.32±2.19^a
MS+sh组		15.61±1.29^b		5.54±1.33^b
MS+OV组		36.97±3.25^b		12.05±2.92^b
F		34.510^＊＊		62.071^＊＊
Control组	57.23±15.76		762.45±82.39		686.45±27.33
MS组	152.49±23.19^a		349.89±48.57^a		293.38±14.81^a
MS+sh组	92.62±18.72^b		568.11±66.12^b		505.03±22.17^b
MS+OV组	261.83±27.35^b		210.43±30.72^b		186.53±9.29^b
F	48.822^＊＊		31.743^＊＊		25.511^＊＊

组别	Fe²⁺/（μmol/kg）		GSH/（mmol/g）		SOD/（U/mg）
组别		ROS/%		MDA/（μmol/g）
Control组		2.46±0.22		3.15±0.71
MS组		25.76±2.63^a		8.32±2.19^a
MS+sh组		15.61±1.29^b		5.54±1.33^b
MS+OV组		36.97±3.25^b		12.05±2.92^b
F		34.510^＊＊		62.071^＊＊
Control组	57.23±15.76		762.45±82.39		686.45±27.33
MS组	152.49±23.19^a		349.89±48.57^a		293.38±14.81^a
MS+sh组	92.62±18.72^b		568.11±66.12^b		505.03±22.17^b
MS+OV组	261.83±27.35^b		210.43±30.72^b		186.53±9.29^b
F	48.822^＊＊		31.743^＊＊		25.511^＊＊

组别	Collagen Ⅱ	Aggrecan	MMP-13
组别	p53	SLC7A11	GPX4
Control组	0.12±0.03	1.06±0.15	0.95±0.13
MS组	0.75±0.12^a	0.63±0.09^a	0.51±0.08^a
MS+sh组	0.18±0.05^b	0.88±0.11^b	0.72±0.10^b
MS+OV组	1.13±0.16^b	0.31±0.06^b	0.26±0.05^b
F	110.134^＊＊	32.893^＊＊	23.031^＊＊
Control组	1.01±0.14	0.90±0.08	0.23±0.06
MS组	0.46±0.11^a	0.39±0.07^a	0.56±0.09^a
MS+sh组	0.67±0.13^b	0.58±0.09^b	0.39±0.06^b
MS+OV组	0.29±0.08^b	0.14±0.04^b	0.95±0.13^b
F	19.624^＊＊	35.932^＊＊	39.824^＊＊

过度机械应力调控Piezo1介导成软骨细胞铁死亡的机制探讨

The mechanism of excessive mechanical stress modulates Piezo1-mediated ferroptosis in chondrocytes

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