Establishment of Ferroptosis Model in Human Aortic Vascular Smooth Muscle Cells
10.3870/j.issn.1672-0741.24.04.002
- VernacularTitle:人主动脉血管平滑肌细胞铁死亡模型的建立
- Author:
Jiannan YE
1
;
Xiang WEI
;
Xin FENG
Author Information
1. 华中科技大学同济医学院附属同济医院心脏大血管外科,武汉 430030
- Keywords:
aorta;
vascular smooth muscle cell;
ferroptosis
- From:
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
2024;53(4):427-432
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish and validate a ferroptosis model in human aortic vascular smooth muscle cells(HAVSMCs).Methods HAVSMCs were obtained from aortic arch tissues of donors who underwent heart transplantation at the Cardiothoracic and Vascular Surgery Department,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology.Ferroptosis was induced through imidazole ketone erastin(IKE)and cystine-deficient medium(CD)in HAVSMCs.Cell counting kit-8(CCK-8),human lactate dehydrogenase(LDH)assay kit,flow cytometry and cell immunofluores-cence staining were used to evaluate whether ferroptosis was successfully induced in HAVSMCs.Results After the induction of ferroptosis in HAVSMCs for a certain period of time,obvious morphological changes of HAVSMCs were observed under the optical microscope,and the flow cytometry analysis showed that the proportion of propidium iodide(PI)positive stained cells was significantly increased under the induction of IKE/CD.The results of CCK-8 and LDH assays showed that the cell viability of HAVSMCs was significantly reduced,and the level of cell damage was markedly increased under IKE/CD induction.Ferrostatin-1,an inhibitor of ferroptosis,was able to reverse the toxic effects of ferroptosis induction on the cells.The flow cytometry analysis of BODIPY-C11 and immunofluorescence staining of 4-hydroxynonenal(4-HNE)showed a notable increase in lipid peroxidation levels in IKE/CD-induced HAVSMCs.Conclusion In this study,a ferroptosis model was successfully established in human aor-tic vascular smooth muscle cells with IKE and cystine-deficient medium,which provided an experimental basis for subsequent researches.
