Effect of Benzimidazole Compound on Endoplasmic Reticulum Stress and Unfolded Protein Response (UPR) Signaling Pathway in Hydrogen Peroxide (H2O2)-Induced BEAS-2B Cells

Aysel Eraslan Şakar; Meral Urhan Küçük; Ronak Haj Ersan; Öztekin Algül

Makale detayları

Effect of Benzimidazole Compound on Endoplasmic Reticulum Stress and Unfolded Protein Response (UPR) Signaling Pathway in Hydrogen Peroxide (H2O2)-Induced BEAS-2B Cells

Yazarlar:

Aysel Eraslan Şakar | Meral Urhan Küçük | Ronak Haj Ersan | Öztekin Algül

Yayıncı:

Mugla Sitki Kocman University

Ana Başlıklar:

Klinik Tıp Bilimleri (Diğer)

Dergi:

Muğla Sıtkı Koçman Üniversitesi Tıp Dergisi Vol.12 Issue 2, pp.127-134

Belge Tipi:

Makale

Yayın Tarihi:

8/2025

Belge Dili:

İngilizce

Document ID (Org No.):

2001357191

pISSN:

2148-8118

Anahtar kelimeler:

Özet:

 The primary objective of this work was to elucidate the impact of endoplasmic reticulum stress on both mRNA level and protein expression on BEAS-2B cells under oxidative stress conditions. The secondary objective was to assess the effect of RHE 231, a benzimidazole compound, on unfolded protein response signaling pathway. To identify non-toxic doses, cell viability under H2O2 and RHE 231 exposure was evaluated via the MTT assay, while cellular lipid oxidation due to H2O2 was measured using the MDA assay. Quantitative real-time PCR was employed to analyze ATF6, PERK, IRE1α, and GRP78 gene expression levels, while IRE1α and ATF4 protein levels were assessed through western blot method. Malondialdehyde levels were notably higher than the control group at ten and twenty µM concentrations at 24 hours. Significant increases in PERK and IRE1α mRNA levels were detected in the groups in which benzimidazole compound and hydrogen peroxide were applied combined than control group. No significant differences in GRP78 and ATF6 mRNA expression levels were detected in any group relative to the control group. Western blot analysis revealed that ATF4 and IRE1 protein expression was higher in the H2O2 alone and combined with benzimidazole compound groups than control group. In BEAS-2B cells, H2O2-induced oxidative damage led to the activation of ER stress and launch of the unfolded protein response pathway. Although protein levels diminished in certain groups treated with the benzimidazole compound, it was observed that this decrease did not exhibit a dose-dependent pattern.

 The primary objective of this work was to elucidate the impact of endoplasmic reticulum stress on both mRNA level and protein expression on BEAS-2B cells under oxidative stress conditions. The secondary objective was to assess the effect of RHE 231, a benzimidazole compound, on unfolded protein response signaling pathway. To identify non-toxic doses, cell viability under H2O2 and RHE 231 exposure was evaluated via the MTT assay, while cellular lipid oxidation due to H2O2 was measured using the MDA assay. Quantitative real-time PCR was employed to analyze ATF6, PERK, IRE1α, and GRP78 gene expression levels, while IRE1α and ATF4 protein levels were assessed through western blot method. Malondialdehyde levels were notably higher than the control group at ten and twenty µM concentrations at 24 hours. Significant increases in PERK and IRE1α mRNA levels were detected in the groups in which benzimidazole compound and hydrogen peroxide were applied combined than control group. No significant differences in GRP78 and ATF6 mRNA expression levels were detected in any group relative to the control group. Western blot analysis revealed that ATF4 and IRE1 protein expression was higher in the H2O2 alone and combined with benzimidazole compound groups than control group. In BEAS-2B cells, H2O2-induced oxidative damage led to the activation of ER stress and launch of the unfolded protein response pathway. Although protein levels diminished in certain groups treated with the benzimidazole compound, it was observed that this decrease did not exhibit a dose-dependent pattern.

 The primary objective of this work was to elucidate the impact of endoplasmic reticulum stress on both mRNA level and protein expression on BEAS-2B cells under oxidative stress conditions. The secondary objective was to assess the effect of RHE 231, a benzimidazole compound, on unfolded protein response signaling pathway. To identify non-toxic doses, cell viability under H2O2 and RHE 231 exposure was evaluated via the MTT assay, while cellular lipid oxidation due to H2O2 was measured using the MDA assay. Quantitative real-time PCR was employed to analyze ATF6, PERK, IRE1α, and GRP78 gene expression levels, while IRE1α and ATF4 protein levels were assessed through western blot method. Malondialdehyde levels were notably higher than the control group at ten and twenty µM concentrations at 24 hours. Significant increases in PERK and IRE1α mRNA levels were detected in the groups in which benzimidazole compound and hydrogen peroxide were applied combined than control group. No significant differences in GRP78 and ATF6 mRNA expression levels were detected in any group relative to the control group. Western blot analysis revealed that ATF4 and IRE1 protein expression was higher in the H2O2 alone and combined with benzimidazole compound groups than control group. In BEAS-2B cells, H2O2-induced oxidative damage led to the activation of ER stress and launch of the unfolded protein response pathway. Although protein levels diminished in certain groups treated with the benzimidazole compound, it was observed that this decrease did not exhibit a dose-dependent pattern.

 Bu çalışmanın birincil amacı, oksidatif stres koşulları altında endoplazmik retikulum stresinin BEAS-2B hücrelerinde mRNA düzeyleri ile protein ekspresyonu üzerindeki etkilerini aydınlatmaktı. İkincil amacı ise, benzimidazol türevi bileşik olan RHE 231in katlanmamış protein yanıtı sinyal yolağı üzerindeki etkisini değerlendirmekti. Toksik olmayan dozları belirlemek için, hidrojen peroksit H2O2 ve RHE 231 maruziyeti altındaki hücre canlılığı MTT testi ile değerlendirilirken, H2O2nun neden olduğu hücresel lipid oksidasyonu MDA testi ile ölçüldü. ATF6, PERK, IRE1α ve GRP78 gen ekspresyon düzeylerini analiz etmek için kantitatif gerçek zamanlı PCR kullanıldı, IRE1α ve ATF4 protein düzeyleri ise Western blot yöntemiyle değerlendirildi. Malondialdehit MDA seviyeleri, 24 saat sonunda kontrol grubuna kıyasla 10 ve 20 µM konsantrasyonlarında belirgin şekilde daha yüksekti. Benzimidazol türevi bileşik ve H2O2 uygulanan gruplarda, kontrol grubuna kıyasla PERK ve IRE1α mRNA düzeylerinde anlamlı artışlar tespit edildi. GRP78 ve ATF6 mRNA ekspresyon düzeylerinde ise herhangi bir grupta kontrol grubuna göre anlamlı bir fark gözlemlenmedi. Western blot analizleri, ATF4 ve IRE1α protein ekspresyonunun, sadece H2O2 ve benzimidazol bileşiği ile kombinasyon halinde uygulanan gruplarda kontrol grubuna kıyasla daha yüksek olduğunu ortaya koydu. BEAS-2B hücrelerinde, H2O2ye bağlı oksidatif hasar, ER stresinin aktivasyonuna ve katlanmamış protein yanıt yolağının başlatılmasına yol açtı. Benzimidazol türevi bileşik uygulanan belirli gruplarda protein düzeylerinin azaldığı gözlemlense de, bu azalmanın doz bağımlı bir model sergilemediği gözlendi.

Makale detayları

Effect of Benzimidazole Compound on Endoplasmic Reticulum Stress and Unfolded Protein Response (UPR) Signaling Pathway in Hydrogen Peroxide (H2O2)-Induced BEAS-2B Cells

Yazarlar:

Aysel Eraslan Şakar | Meral Urhan Küçük | Ronak Haj Ersan | Öztekin Algül

Yayıncı:

Mugla Sitki Kocman University

Ana Başlıklar:

Klinik Tıp Bilimleri (Diğer)

Dergi:

Muğla Sıtkı Koçman Üniversitesi Tıp Dergisi Vol.12 Issue 2, pp.127-134

Belge Tipi:

Makale

Yayın Tarihi:

8/2025

Belge Dili:

İngilizce

Document ID (Org No.):

2001357191

pISSN:

2148-8118

BEAS-2B

Benzimidazole

ER Stress

Hydrogen Peroxide

Oxidative StressBEAS-2B

Benzimidazol

ER Stresi

Hidrojen Peroksit

Oksidatif Stres

Sayı

Makale detayları

Yazarlar: Aysel Eraslan Şakar | Meral Urhan Küçük | Ronak Haj Ersan | Öztekin Algül

Yayıncı: Mugla Sitki Kocman University

Ana Başlıklar: Klinik Tıp Bilimleri (Diğer)

Dergi: Muğla Sıtkı Koçman Üniversitesi Tıp Dergisi Vol.12 Issue 2, pp.127-134

Belge Tipi: Makale

Yayın Tarihi: 8/2025

Belge Dili: İngilizce

Document ID (Org No.): 2001357191

pISSN: 2148-8118

Sayı

Yazarlar:

Aysel Eraslan Şakar | Meral Urhan Küçük | Ronak Haj Ersan | Öztekin Algül

Yayıncı:

Mugla Sitki Kocman University

Ana Başlıklar:

Klinik Tıp Bilimleri (Diğer)

Dergi:

Muğla Sıtkı Koçman Üniversitesi Tıp Dergisi Vol.12 Issue 2, pp.127-134

Belge Tipi:

Makale

Yayın Tarihi:

8/2025

Belge Dili:

İngilizce

Document ID (Org No.):

2001357191

pISSN:

2148-8118