中国组织工程研究 ›› 2024, Vol. 28 ›› Issue (20): 3202-3208.doi: 10.12307/2024.334

• 骨组织构建 bone tissue construction • 上一篇    下一篇

氧化应激促进成骨细胞铁死亡介导激素性股骨头坏死的病理过程

章家皓1,刘予豪2,周  驰2,莫  亮1,方汉军2,陈镇秋2   

  1. 1广州中医药大学,广东省广州市  510405;2 广州中医药大学第一附属医院,广东省广州市  510405
  • 收稿日期:2023-04-23 接受日期:2023-06-06 出版日期:2024-07-18 发布日期:2023-09-11
  • 通讯作者: 陈镇秋,博士,主任中医师,广州中医药大学第一附属医院,广东省广州市 510405 刘予豪,博士,主治中医师,广州中医药大学第一附属医院,广东省广州市 510405
  • 作者简介:章家皓,男,1996年生,广东省揭阳市人,汉族,广州中医药大学在读硕士,医师,主要从事中医药防治骨关节疾病的研究。
  • 基金资助:
    广州中医药大学“双一流”与高水平大学学科协同创新团队项目 (2021xk46),项目负责人:陈镇秋;国家自然科学基金项目(82104883),项目负责人:刘予豪;广东省中医药局科研项目(20231162),项目负责人:陈镇秋;国家中医药传承创新中心科研专项(2022QN07),项目负责人:方汉军

The pathological progression of steroid-induced osteonecrosis of the femoral head caused by oxidative stress-induced osteoblast ferroptosis

Zhang Jiahao1, Liu Yuhao2, Zhou Chi2, Mo Liang1, Fang Hanjun2, Chen Zhenqiu2   

  1. 1Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; 2First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • Received:2023-04-23 Accepted:2023-06-06 Online:2024-07-18 Published:2023-09-11
  • Contact: Chen Zhenqiu, MD, Chief physician, First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China Liu Yuhao, MD, Attending physician, First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • About author:Zhang Jiahao, Master, Physician, Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • Supported by:
    “Double First-class” and High-level University Discipline Collaborative Innovation Team Project of Guangzhou University of Chinese Medicine, No. 2021xk46 (to CZQ); National Natural Science Foundation of China, No. 82104883 (to LYH); Guangdong Provincial Bureau of Chinese Medicine Research Project, No. 20231162 (to CZQ); National Center for Traditional Chinese Medicine Inheritance and Innovation Research Special Project, No. 2022QN07 (to FHJ)

摘要:


文题释义:

氧化应激:指体内氧化与抗氧化作用失衡的一种状态,倾向于氧化,导致中性粒细胞炎性浸润,蛋白酶分泌增加,产生大量氧化中间产物。
铁死亡:由于铁代谢功能障碍,细胞内铁过量导致脂质活性氧自由基生产过多,进而发生一系列的酶促反应,大量铁依赖性的脂质氢过氧化物积累,破坏细胞膜结构与功能,进而引发细胞死亡的细胞死亡形式。


背景:研究表明,糖皮质激素诱发股骨头坏死过程中骨代谢失衡与氧化应激密切相关。

目的:探究氧化应激诱导成骨细胞铁死亡、促进成骨细胞凋亡、参与激素性股骨头坏死的病理机制。
方法:收集47例激素性股骨头坏死患者的一般资料、血清标本和6例股骨头标本,根据ARCO分期标准将血清标本分为ARCOⅡ期、Ⅲ期及Ⅳ期,股骨头标本分为ARCO Ⅲ期及Ⅳ期。检测血清中丙二醛、超氧化物歧化酶1水平;通过DIA蛋白定量测序、免疫蛋白印迹试验及碱性磷酸检测并验证患者股骨头中超氧化物歧化酶1、谷胱甘肽过氧化物酶4、Bcl-2蛋白的表达。

结果与结论:①激素性股骨头坏死患者的ARCO分期与年龄、性别、侧别无关;②ARCO Ⅲ期患者血清中丙二醛、超氧化物歧化酶1水平较Ⅱ期、Ⅳ期高;③在坏死区域与正常区域对比中,DIA蛋白定量测序结果显示,差异蛋白的功能主要与氧化还原相关,其中坏死区域中超氧化物歧化酶1、谷胱甘肽过氧化物酶4、Bcl-2蛋白表达较正常区域低,且ARCO Ⅳ期低于Ⅲ期;Western blot验证了DIA蛋白定量结果,且坏死区域中碱性磷酸酶活性低于正常区域,Ⅳ期低于Ⅲ期;④在坏死区域与硬化带区域中,差异蛋白的功能同样与氧化还原相关,且坏死区域中超氧化物歧化酶1、谷胱甘肽过氧化物酶4、Bcl-2蛋白表达及碱性磷酸酶活性较硬化带区域低,且Ⅳ期低于Ⅲ期;⑤结果说明,糖皮质激素可通过上调氧化应激水平,诱导成骨细胞铁死亡的发生,抑制成骨功能,影响激素性股骨头坏死的进展。

https://orcid.org/0009-0002-5701-7770(章家皓)

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程

关键词: 氧化应激, 铁死亡, 成骨细胞, 激素性股骨头坏死, SOD1, Gpx4

Abstract: BACKGROUND: Studies have shown that imbalance of bone metabolism during glucocorticoid-induced osteonecrosis of the femoral head necrosis is closely related to oxidative stress.
OBJECTIVE: To investigate the pathological mechanism by which oxidative stress-induced ferroptosis promote apoptosis in osteoblasts involved in steroid-induced osteonecrosis of the femoral head.
METHODS: General data and serum specimens were collected from 47 patients with steroid-induced osteonecrosis of the femoral head. In addition, six femoral head specimens were collected from these patients. According to the Association Research Circulation Osseous (ARCO) staging system, serum specimens were grouped into ARCO II, III, and IV, while femoral head specimens were classified into ARCO III and IV. Serum levels of malondialdehyde and superoxide dismutase 1 were measured. The protein expression of superoxide dismutase 1, glutathione peroxidase 4, Bcl-2 in the femoral head was detected and verified by Data independent acquisition (DIA) for quantitative sequencing, western blot and alkaline phosphate detection.
RESULTS AND CONCLUSION: The ARCO stage of patients with steroid-induced osteonecrosis of the femoral head was independent of age, sex and necrotic side. The serum levels of malondialdehyde and superoxide dismutase 1 were higher in patients with ARCO stage III compared with those with ARCO stage II and IV. The results of DIA protein quantification showed that the function of differential proteins was mainly related to redox. The levels of superoxide dismutase 1, glutathione peroxidase 4, and Bcl-2 in the necrotic region were lower than in the normal region, as well as lower in ARCO stage IV than in ARCO stage III. Western blot verified the results of DIA protein quantification. The alkaline phosphatase activity was lower in the necrotic region than in the normal region, as well as lower in ARCO stage IV than in ARCO stage III. In the necrotic and sclerotic regions, the function of differential proteins was also related to redox, and superoxide dismutase 1, glutathione peroxidase 4, Bcl-2 protein expression and alkaline phosphatase activity were lower in the necrotic area than in the sclerotic region, as well as lower in ARCO stage IV than in ARCO stage III. To conclude, glucocorticoids can influence the progression of steroid-induced osteonecrosis of the femoral head by upregulating oxidative stress levels, inducing osteoblast ferroptosis, and inhibiting osteogenic function.

Key words: oxidative stress, ferroptosis, osteoblast, steroid-induced osteonecrosis of the femoral head, SOD1, Gpx4

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