叔丁基氢过氧化物可诱导髓核细胞发生铁死亡
tert-Butyl hydroperoxide can induce ferroptosis in nucleus pulposus cells
摘要
Abstract
BACKGROUND:Degeneration of nucleus pulposus cells is a key component of intervertebral disc degeneration.Ferroptosis,a novel form of programmed cell death,is closely associated with the onset and progression of intervertebral disc degeneration;however,its precise mechanisms remain unclear. OBJECTIVE:To establish an oxidative stress model in vitro by inducing ferroptosis in nucleus pulposus cells using tert-butyl hydroperoxide and to investigate the mechanisms of tert-butyl hydroperoxide-induced ferroptosis in nucleus pulposus cells,thereby elucidating the role of ferroptosis in the pathogenesis of intervertebral disc degeneration. METHODS:Nucleus pulposus cells were treated with varying concentrations of tert-butyl hydroperoxide(0,25,50,100,and 200 μmol/L),and cell morphology and viability were assessed using fluorescence microscopy and the cell counting kit-8 assay.Interventions with 100 μmol/L tert-butyl hydroperoxide,10 μmol/L RSL3,or dimethylsulfoxide were applied to nucleus pulposus cells,and cell proliferation was evaluated using the EdU assay.The expression levels of ferroptosis-related proteins(glutathione peroxidase 4,ferritin heavy chain 1,PTGS2,and ACSL4)and intervertebral disc degeneration marker proteins(matrix metalloproteinase 13 and Col2A)were analyzed via western blot and immunofluorescence.Additionally,reactive oxygen species and lipid peroxidation levels were quantified using the reactive oxygen species detection kit and C11-BODIPY probe.Mitochondrial morphological changes were observed under transmission electron microscopy. RESULTS AND CONCLUSION:(1)tert-Butyl hydroperoxide treatment significantly reduced the viability and proliferation of nucleus pulposus cells.(2)tert-Butyl hydroperoxide induced typical ferroptosis-related morphological changes in nucleus pulposus cells.(3)tert-Butyl hydroperoxide exposure led to a decrease in the expression of ferroptosis-suppressing proteins glutathione peroxidase 4 and ferritin heavy chain 1,while increasing the expression of ferroptosis-promoting factors ACSL4 and PTGS2.(4)tert-Butyl hydroperoxide elevated intracellular reactive oxygen species production and lipid peroxidation levels in nucleus pulposus cells.(5)Transmission electron microscopy revealed ferroptosis-specific mitochondrial changes in nucleus pulposus cells treated with tert-butyl hydroperoxide,including contraction,reduced cristae,and increased membrane density.(6)tert-Butyl hydroperoxide treatment also resulted in the increased expression of matrix metalloproteinase 13 and decreased expression of Col2A in nucleus pulposus cells.In conclusion,tert-butyl hydroperoxide induces ferroptosis in nucleus pulposus cells,contributing to the development of intervertebral disc degeneration.This process may represent a key pathological mechanism in intervertebral disc degeneration and offers potential targets for developing novel therapeutic strategies.关键词
TBHP/铁死亡/髓核细胞/氧化应激/椎间盘退变Key words
tert-butyl hydroperoxide/ferroptosis/nucleus pulposus cells/oxidative stress/intervertebral disc degeneration分类
医药卫生引用本文复制引用
谌超,张帆,胡耀全,吕正品,何其聪,杨阳子久,罗皓严,吴贵帅,左乾林,王学楠..叔丁基氢过氧化物可诱导髓核细胞发生铁死亡[J].中国组织工程研究,2025,29(32):6858-6865,8.基金项目
国家自然科学基金项目(82160428),项目负责人:张帆 （82160428）
昆明医科大学第一附属医院535人才项目(2022535D10),项目负责人:张帆 （2022535D10）
云南省"兴滇英才支持计划"医疗卫生人才培养计划项目,项目负责人:张帆 （）
昆明医科大学中青年学科带头人及后备人选-"乘风人才"培养计划项目(J13397034),项目负责人:张帆 （J13397034）
昆明医科大学一流学科团队(2024XKTDYS05),项目参与人:张帆 the National Natural Science Foundation of China,No.82160428(to ZF) （2024XKTDYS05）
535 Talent Project of First Affiliated Hospital of Kunming Medical University,No.2022535D10(to ZF) （to ZF）
Yunnan Province"Xingdian Talent Support Plan"Project for Medical and Health Talent Training(No project number available)(to ZF) （No project number available）
The"Rising Star"Talent Training Program for Young and Middle-Aged Discipline Leaders and Reserve Candidates at Kunming Medical University,No.J13397034(to ZF) （to ZF）
First-class Academic Team of Kunming Medical University,No.2024XKTDYS05(to ZF[project participant]) （to ZF[project participant]）
