Investigation of Possible Protective Effects of Resveratrol on Oxidative Stress and Ferroptosis in PFOA Exposure in HepG-2 Cells

Year 2024, Volume: 16 Issue: 3, 283 - 292, 29.10.2024

Didem Oral , Ceyhan Hacıoğlu

https://doi.org/10.18521/ktd.1464261

Abstract

Objective: Exposure to perfluorooctanoic acid (PFOA) is linked to adverse health effects, including cancer and hepatic diseases. PFOA induces reactive oxygen species generation in human hepatic cells, causing oxidative stress and cell death. Resveratrol (RSV) has garnered attention for its protective effects against xenobiotic-induced damage, yet its impact on PFOA-induced oxidative stress and ferroptosis in the liver remains understudied. This study investigates RSV's protective mechanisms against oxidative stress and ferroptosis in HepG2 cells exposed to PFOA.
Method: HepG2 cells were cultured in DMEM with 10% FBS and 1% penicillin/streptomycin in a 5% CO2 incubator at 37°C. PFOA was added to the cells at concentrations ranging from 0 to 450 µM and incubated at 37°C for 24 hours. The IC50 was determined to be 457 μM. To examine RSV's protective effects, cells were treated with 60 μM RSV. Following treatment with PFOA, RSV, and the combination of PFOA+RSV, cell lysates were prepared for analysis. Oxidative stress and ferroptosis parameters were measured spectrophotometrically using ELISA.
Results: In the PFOA+RSV group, antioxidant capacity increased, and ferroptosis was suppressed compared to the control. Conversely, the PFOA group showed decreased antioxidant capacity, increased oxidant capacity, and induced ferroptosis compared to the control and RSV-treated groups.
Conclusion: PFOA exposure heightens oxidative stress and ferroptosis, whereas RSV treatment significantly reduces hepatic oxidative stress and protects against ferroptosis during PFOA exposure.

Keywords

PFOA, Resveratrol, Oxidative stress, HepG2, Ferroptosis

Ethical Statement

Etik kurul onayı gerekli değildir

Supporting Institution

Duzce University,he University of Duzce Scientific Research Unit Rapid Support

Project Number

The funding support for this study was provided by the University of Duzce Scientific Research Unit Rapid Support (Project No: 2022.14.02.1358).

PFOA Maruziyetinde Resveratrol’ün Oksidatif Stres ve Ferroptosis Üzerindeki Olası Koruyucu Etkilerinin HepG2 Hücrelerinde Araştırılması

Abstract

Amaç: Perflorooktanoik asit (PFOA) maruziyeti, kanser ve karaciğer hastalıkları dahil olmak üzere çeşitli olumsuz sağlık etkileri ile ilişkilendirilmiştir. PFOA, insan karaciğer hücrelerinde reaktif oksijen türlerinin oluşumunu indükleyerek oksidatif strese ve hücre ölümüne neden olur. Son yıllarda, Resveratrolün (RSV) ksenobiyotiklerin neden olduğu hasarlara karşı koruyucu etkisi üzerine çalışmalar önem kazanmıştır. Ancak, RSV'nin karaciğerde PFOA'nın neden olduğu oksidatif stres ve ferroptozis üzerindeki koruyucu etkileri hakkında yeterli çalışma bulunmamaktadır. Bu çalışmada, PFOA maruziyeti sonucu HepG2 hücrelerinde oluşabilecek oksidatif stres ve ferroptozis üzerine RSV'nin koruyucu etki mekanizmalarını araştırmayı amaçladık.
Metod: HepG2 hücreleri, %5 CO2 inkübatöründe 37°C'de %10 FBS ve %1 penisilin/streptomisin içeren DMEM'de kültürlendi. PFOA, 0-450 µM konsantrasyonlarında hücrelere eklendi ve 37°C'de 24 saat inkübe edildi. IC50, 457 μM olarak belirlendi. RSV'nin koruyucu etkilerini değerlendirmek için hücreler 60 μM RSV ile muamele edildi. PFOA, RSV ve PFOA+RSV ile muamele sonrasında, hücre lizatı hazırlandı ve analizler için kullanıldı. Oksidatif stres ve ferroptozis parametreleri, ELISA yöntemiyle spektrofotometrik olarak belirlendi.
Bulgular: PFOA+RSV grubunda, kontrol grubuna kıyasla antioksidan kapasite artmış ve ferroptozis baskılanmıştır. Buna karşılık, PFOA grubunda antioksidan kapasite azalmış, oksidan kapasite artmış ve kontrol ve RSV ile muamele edilen gruplara kıyasla ferroptozis indüklenmiştir.
Sonuç: PFOA maruziyeti oksidatif stresi ve ferroptozisi artırırken, RSV tedavisi karaciğer oksidatif stresini önemli ölçüde azaltır ve PFOA maruziyeti sırasında ferroptozise karşı korur.

Keywords

PFOA, Resveratrol, Oksidatif Stres, HepG2, Ferroptoz

Project Number

The funding support for this study was provided by the University of Duzce Scientific Research Unit Rapid Support (Project No: 2022.14.02.1358).

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