Research on the mechanism of cytochrome P4502C regulating ferroptosis in hu-man retinal microvascular endothelial cells under high-glucose conditions through the GPX4/ACSL4 axis
10.13389/j.cnki.rao.2025.0119
- VernacularTitle:细胞色素P4502C通过GPX4/ACSL4轴调控高糖环境下人视网膜微血管内皮细胞铁死亡的机制研究
- Author:
Ting LUO
1
;
Yan LI
;
Like XIE
;
Hongbin LÜ
Author Information
1. 646000 四川省泸州市,西南医科大学附属医院眼科;641499 四川省成都市,简阳市人民医院眼科
- Publication Type:Journal Article
- Keywords:
diabetic retinopathy;
cytochrome P4502C;
rifampicin;
fenofibrate;
ferroptosis;
GPX4/ACSL4 pathway
- From:
Recent Advances in Ophthalmology
2025;45(9):691-695
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the molecular mechanisms by which cytochrome P4502C regulates ferroptosis in hu-man retinal microvascular endothelial cells(hRMECs)under high-glucose conditions.Methods Human retinal microvas-cular endothelial cells(hRMECs)were cultured in vitro and divided into six groups:the NG group(cells cultured in ser-um-free DMEM medium containing 5.5 mmol·L-1 glucose);the NG+fenofibrate group(10 μmol·L-1 fenofibrate added to the NG group);the NG+rifampicin group(10 μmol·L-1 rifampicin added to the NG group);the HG group(cells cul-tured in serum-free DMEM medium containing 25.5 mmol·L-1 glucose);the HG+fenofibrate group(10 μmol·L-1 feno-fibrate added to the HG group);and the HG+rifampicin group(10 μmol·L-1 rifampicin added to the HG group).After 48 hours of culture,subsequent experiments were conducted.Cell viability was assessed using the CCK-8 assay.The levels of malondialdehyde(MDA)and iron content in hRMECs were measured using commercial assay kits.The relative fluores-cence intensity of Liperfluo was quantified by flow cytometry.The mRNA expression levels of glutathione peroxidase 4(GPX4),cytochrome P4502C,and acyl-CoA synthetase long-chain family member 4(ACSL4)in hRMECs were analyzed using real-time quantitative PCR.The protein expression levels of GPX4,cytochrome P4502C,and ACSL4 in hRMECs were evaluated using Western blot analysis.Results Optimal concentration analysis showed that the 10 μmnol·L-1 fenofibrate group had the highest cell viability compared to the HG group(P＜0.05),while the 10 μmol·L-1 rifampicin group had the lowest cell viability(P＜0.05).Thus,10 μmol·L-1 was selected as the optimal concentration for both fenofibrate and rif-ampicin for subsequent experiments.Cell viability in the NG,NG+fenofibrate,NG+rifampicin,HG,HG+fenofibrate,and HG+rifampicin groups was(100.00±7.85)％,(102.15±9.07)％,(69.97±4.22)％,(61.74±6.13)％,(83.64±7.58)％,and(56.96±5.34)％,respectively.Compared with the NG group,the HG group had significantly lower cell via-bility(P＜0.05).Compared with the HG group,the HG+fenofibrate group had significantly higher cell viability(P＜0.05).Compared with the NG group,the HG group had higher levels of MDA,iron content,and Liperfluo relative fluores-cence intensity(all P＜0.05).Compared with the HG group,the HG+fenofibrate group had lower levels of MDA,iron content,and Liperfluo relative fluorescence intensity(all P＜0.05);the HG+rifampicin group had higher levels of these parameters(all P＜0.05).Compared with the NG group,the HG group had higher mRNA and protein expression levels of cytochrome P4502C and ACSL4,but lower levels of GPX4 mRNA and protein(all P＜0.05).Compared with the HG group,the HG+fenofibrate group had lower mRNA and protein expression levels of cytochrome P4502C and ACSL4,but higher lev-els of GPX4 mRNA and protein(all P＜0.05);the HG+rifampicin group had higher mRNA and protein expression levels of cytochrome P4502C and ACSL4,but lower levels of GPX4 mRNA and protein(all P＜0.05).Conclusion Cytochrome P4502C can promote ferroptosis of hRMECs under high-glucose conditions by regulating the GPX4/ACSL4 signaling axis;in-hibition of cytochrome P4502C expression can exert a protective effect on DR;modulation of the expression levels or bal-ance of the key factors GPX4 and ACSL4 in this pathway may hold promise as a potential strategy to slow the progression of DR.
