Investigation of The Effects of Carvacrol on Lambda-Cyhalothrin-Induced Gastrointestinal Injury in Rats: Modulation of Apoptosis and Endoplasmic Reticulum Stress Pathways

Öz

This study investigated the mechanisms associated with oxidative stress, apoptosis, and endoplasmic reticulum (ER) stress in gastrointestinal damage induced by lambda-cyhalothrin (λ-CHL) in rats and evaluated the protective effects of carvacrol (CRV) against this toxicity. Rats were divided into five groups: control, CRV, λ-CHL, λ-CHL+CRV25, and λ-CHL+CRV50, and treatments were administered orally for 21 days. In the λ-CHL group, malondialdehyde (MDA) levels significantly increased, whereas glutathione peroxidase (GPx), superoxide dismutase (SOD), and glutathione (GSH) levels decreased (p<0.001). Additionally, apoptotic signaling pathways were activated by a pro-apoptotic shift in the Bcl2-associated X protein (Bax)/ B-cell lymphoma 2 (Bcl2) ratio and increased Casp3 activation (p<0.001). Moreover, endoplasmic reticulum (ER) stress markers, including activating transcription factor 6 (Atf6), C/EBP homologous protein (Chop), and protein kinase RNA-like endoplasmic reticulum kinase (Perk), were significantly elevated (p<0.001). In the CRV-treated groups, particularly in the λ-CHL+CRV50 group, MDA levels were significantly suppressed (p<0.001), while antioxidant defense parameters such as GPx, SOD activity, and GSH levels were significantly increased (p<0.001). Furthermore, Bcl2 was elevated, whereas Bax, Casp3, and ER stress markers were significantly reduced (p<0.001). CRV alone showed patterns similar to the control group. In conclusion, λ-CHL induces gastrointestinal damage through multiple molecular mechanisms involving oxidative stress, apoptosis, and ER stress, whereas CRV exhibits a dose-dependent protective effect against this toxicity.

Anahtar Kelimeler

• Carvacrol
• Endoplasmic reticulum stress
• Gastrointestinal injury
• Lambda-cyhalothrin
• Oxidative stress

Etik Beyan

Bu çalışma, Atatürk Üniversitesi Hayvan Deneyleri Yerel Etik Kurulu’nun 25.02.2026 tarih ve 2026/2 sayılı, 8 nolu kararı ile onaylanmış olup, tüm deneysel uygulamalar ulusal ve uluslararası etik kurallar çerçevesinde gerçekleştirilmiştir.

Sıçanlarda Lambda-Sihalotrin Kaynaklı Gastrointestinal Hasar Üzerine Karvakrol’ün Etkilerinin İncelenmesi: Apoptoz ve Endoplazmik Retikulum Stresi Yolaklarının Düzenlenmesi

Öz

Bu çalışmada, sentetik bir Tip II piretroid olan lambda-sihalotrinin (λ-SHL) sıçanlarda oluşturduğu gastrointestinal hasarın oksidatif stres, apoptoz ve endoplazmik retikulum (ER) stresi ile ilişkili mekanizmaları araştırılmış ve karvakrolün (KRV) bu toksisiteye karşı koruyucu etkileri değerlendirilmiştir. Erkek Sprague-Dawley sıçanlar; sağlıklı kontrol, KRV, λ-SHL, λ-SHL+KRV25 ve λ-SHL+KRV50 olmak üzere beş gruba ayrılarak 21 gün süreyle oral yolla uygulanmıştır. λ-SHL grubunda mide ve bağırsak dokularında malondialdehit (MDA) düzeylerinin anlamlı şekilde arttığı, glutatyon peroksidaz (GPx), süperoksit dismutaz (SOD) aktivitesi ve glutatyon (GSH) düzeylerinin ise belirgin olarak azaldığı saptanmıştır (p<0.001). Aynı grupta apoptotik sinyal yolaklarında Bcl2 ile ilişkili X proteini (Bax)/ B-hücreli lenfoma 2 (Bcl2) oranının pro-apoptotik yönde değiştiği ve Casp3 aktivasyonunun yükseldiği belirlenmiştir (p<0.001). Ayrıca endoplazmik retikulum (ER) stresi belirteçleri olan aktive edici transkripsiyon faktörü 6 (Atf6), C/EBP homolog proteini (Chop) ve protein kinaz RNA benzeri endoplazmik retikulum kinazı (Perk) gen ekspresyonlarının anlamlı düzeyde arttığı gözlenmiştir (p<0.001). KRV uygulanan gruplarda, özellikle yüksek doz KRV verilen λ-SHL+KRV50 grubunda, oksidatif stres parametrelerinin bir göstergesi olan MDA seviyelerinin baskılandığı (p<0.001), antioksidan savunma parametrelerinden olan GPx, SOD aktiviteleri ve GSH seviyelerinin ise arttığı tespit edilmiştir (p<0.001). Apoptoz belirteçlerinden olan Bcl2 gen ekspresyon düzeyinin arttığı, Bax ve Casp3 gen ekspresyon seviyeleri ile ER stresi belirteçlerinin anlamlı şekilde gerilediği tespit edilmiştir (p<0.001). Sadece KRV verilen gruplarda ise incelenen parametreler kontrol ve tedavi gruplarına paralel şekilde seyretmiştir. Sonuç olarak, λ-SHL’nin gastrointestinal dokuda çoklu moleküler mekanizmalar üzerinden hasara yol açtığı, KRV’nin ise bu toksisiteye karşı belirgin ve doza bağlı koruyucu etki gösterdiği ortaya konmuştur.

Anahtar Kelimeler

• Endoplazmik retikulum stresi
• Gastrointestinal hasar
• Karvakrol
• Lambda-sihalotrin
• Oksidatif stres

Etik Beyan

Bu çalışma, Atatürk Üniversitesi Hayvan Deneyleri Yerel Etik Kurulu’nun 25.02.2026 tarih ve 2026/2 sayılı, 8 nolu kararı ile onaylanmış olup, tüm deneysel uygulamalar ulusal ve uluslararası etik kurallar çerçevesinde gerçekleştirilmiştir.

Kaynakça

1. Adhikari S, Sarkar B, Chattopadhyay A et al. (2006). Effect of cypermethrin on breeding performance of a freshwater fish, Labeo rohita (Hamilton). Chem Ecol, 22, 211–218.
2. Aksu EH, Kandemir FM, Altun S et al. (2016). Ameliorative effect of carvacrol on cisplatin-induced reproductive damage in male rats. J Biochem Mol Toxicol, 30, 513–520.
3. Bolat M, Çelebi F, Şengül E et al. (2025). Investigation of the protective effect of beta-caryophyllene against indomethacin-induced gastric ulcer in rats: In vivo and in vitro study. Naunyn Schmiedebergs Arch Pharmacol, 1–19.
4. Bozkurt I, Halici Z, Zirh EB, Palabiyik-Yucelik SS (2024). EGCG alleviates ochratoxin A-induced pyroptosis in rat’s kidney by inhibiting NLRP3/Caspase-1/GSDMD signaling pathway. Food Chem Toxicol, 182, 115006.
5. Chen X, Shi C, He M, Xiong S, Xia X (2023). Endoplasmic reticulum stress: molecular mechanism and therapeutic targets. Signal Transduct Target Ther, 8, 402.
6. Da Silva Lima M, Quintans-Júnior LJ, de Santana WA et al. (2013). Anti-inflammatory effects of carvacrol: evidence for a key role of interleukin-10. Eur J Pharmacol, 699, 112–117.
7. Dhouibi, R, Affes, H, Salem, MB et al. (2021). Protective effect of Urtica dioica in induced neurobehavioral changes, nephrotoxicity and hepatotoxicity after chronic exposure to potassium bromate in rats’, Environ. Pollut., 287, p. 117657.
8. Dogan T, Onay E, Yildirim BA (2023). Investigation of the effect of Polygonum cognatum Meissn.ethanol extract on Bax, Caspase-3, Bcl-2, NF-κB and NRF-2/HO-1 pathways in streptozotocin-induced diabetic rats. Diabetes, 5(7), 8.

9. El-Sayed Em, Abd-Allah Ar, Mansour Am, El-Arabey Aa (2015). Thymol and carvacrol prevent cisplatin-induced nephrotoxicity by abrogation of oxidative stress, inflammation, and apoptosis in rats. J Biochem Mol Toxicol, 29, 165–172.
10. Fang T, Tian G, Chen D et al. (2024). Endoplasmic reticulum stress contributes to intestinal injury in intrauterine growth restriction newborn piglets. Animals, 14(18), 2677.
11. Fetoui H, Garoui EM, Zeghal N (2009). Lambda-cyhalothrin-induced biochemical and histopathological changes in the liver of rats: ameliorative effect of ascorbic acid. Exp Toxicol Pathol, 61, 189–196.
12. Garud A, Pawar S, Patil MS, Kale SR, Patil S (2024). A scientific review of pesticides: Classification, toxicity, health effects, sustainability, and environmental impact. Cureus, 16(8), e67945.
13. Hotta M, Nakata R, Katsukawa M et al. (2010). Carvacrol, a component of thyme oil, activates PPARα and γ and suppresses COX-2 expression. J Lipid Res, 51, 132–139.
14. Hsu MC, Wang HT ve Chen CY (2025). Carvacrol protects intestinal epithelial cells from oxidative stress. Int J Mol Sci, 26(8), 3495.
15. Ileriturk M, Kandemir FM (2023). Carvacrol protects against λ-cyhalothrin-induced hepatotoxicity and nephrotoxicity by modulating oxidative stress, inflammation, apoptosis, endoplasmic reticulum stress, and autophagy. Environ Toxicol, 38(7), 1535–1547.
16. Ileriturk M, Kandemir O, Akaras N et al. (2023). Hesperidin has a protective effect on paclitaxel-induced testicular toxicity through regulating oxidative stress, apoptosis, inflammation and endoplasmic reticulum stress. Reprod Toxicol, 108369.
17. Kale M, Rathore N, John S, Bhatnagar D (1999). Lipid peroxidative damage on pyrethroid exposure and alterations in antioxidant status in rat erythrocytes: a possible involvement of reactive oxygen species. Toxicol Lett, 105, 197–205.
18. Kandemir Ö, İleritürk M, Gür C et al. (2025). Carvacrol coadministration ameliorates lambda-cyhalothrin-induced toxicity. J Biochem Mol Toxicol, e70400.
19. Kankılıç NA, Şimşek H, Akaras N et al. (2024). Chrysin ameliorates bortezomib-induced nephrotoxicity in rats by reducing oxidative stress, endoplasmic reticulum stress, inflammatory injury, apoptotic and autophagic cell death. Food Chem Toxicol, 190, 114791.
20. Karaarslan T, Alat O, Can I et al. (2026). Protocatechuic acid mitigates diazinon-induced lung injury in rats through modulation of oxidative stress, inflammatory, Keap-1/Nrf-2/HO-1 and ER stress-mediated apoptotic pathways. Iran J Basic Med Sci, 29(2), 261–270.
21. Livak KJ, Schmittgen TD (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods, 25(4), 402–408.
22. Lopez-Torres B, Ares I, Martínez M et al. (2022). Lambda-cyhalothrin-induced neurotoxicity: alterations in monoaminergic systems and dopaminergic and serotonergic pathways in rat brain. Food Chem Toxicol, 169, 113434.
23. Nafees S, Ahmad ST, Arjumand W et al. (2013). Carvacrol ameliorates thioacetamide-induced hepatotoxicity by abrogation of oxidative stress, inflammation, and apoptosis in liver of Wistar rats. Hum Exp Toxicol, 32, 1292–1304.
24. Oliveira IS, da Silva FV, Viana AFS et al. (2012). Gastroprotective activity of carvacrol on experimentally induced gastric lesions in rodents. Naunyn Schmiedebergs Arch Pharmacol, 385(9), 899–908.
25. Prasanthi K, Rajini P (2005). Fenvalerate-induced oxidative damage in rat tissues and its attenuation by dietary sesame oil. Food Chem Toxicol, 43, 299–306.
26. Sakr S, Rashad WA (2023). Lambda-cyhalothrin-induced pancreatic toxicity in adult albino rats. Sci Rep, 13(1), 11562.
27. Sedlak J, Lindsay RH (1968). Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman’s reagent. Anal Biochem, 25, 192–205.
28. Sharifi-Rad M, Varoni EM, Iriti M et al. (2018). Carvacrol and human health: a comprehensive review. Phytother Res, 32, 1675–1687.
29. Shoorei H, Khaki A, Khaki AA et al. (2019). The ameliorative effect of carvacrol on oxidative stress and germ cell apoptosis in testicular tissue of adult diabetic rats. Biomed Pharmacother, 111, 568–578.
30. Soderlund DM, Clark JM, Sheets LP et al. (2002). Mechanisms of pyrethroid neurotoxicity: implications for cumulative risk assessment. Toxicology, 171, 3–59.
31. Somensi N, Rabelo TK, Guimaraes AG et al. (2019). Carvacrol suppresses LPS-induced pro-inflammatory activation in RAW 264.7 macrophages through ERK1/2 and NF-κB pathway. Int Immunopharmacol, 75, 105743.
32. Sun Y, Oberley LW, Li Y (1988). A simple method for clinical assay of superoxide dismutase. Clin Chem, 34, 497–500.
33. Suntres ZE, Coccimiglio J, Alipour M (2015). The bioactivity and toxicological actions of carvacrol. Crit Rev Food Sci Nutr, 55, 304–318.
34. Şimşek H, Akaras N, Gür C, Küçükler S, Kandemir FM (2023). Beneficial effects of chrysin on cadmium-induced nephrotoxicity in rats: modulating the levels of Nrf2/HO-1, RAGE/NLRP3, and Caspase-3/Bax/Bcl-2 signaling pathways. Gene, 979, 147502.
35. Taslimi P, Caglayan C, Gulcin İ (2017). The impact of some natural phenolic compounds on carbonic anhydrase, acetylcholinesterase, butyrylcholinesterase, and α-glycosidase enzymes: an antidiabetic, anticholinergic, and antiepileptic study. J Biochem Mol Toxicol, 31, e21995.
36. Turgut C, Ornek H, Cutright TJ (2011). Determination of pesticide residues in Turkey’s table grapes: the effect of integrated pest management, organic farming, and conventional farming. Environ Monit Assess, 173, 315–323.
37. Xu J, Zhou F, Ji BP, Pei RS, Xu N (2008). The antibacterial mechanism of carvacrol and thymol against Escherichia coli. Lett Appl Microbiol, 47, 174–179.
38. Xu X, Yu Y, Ling M et al. (2023). Oxidative stress and mitochondrial damage in lambda-cyhalothrin toxicity: A comprehensive review of antioxidant mechanisms. Environ Pollut, 122694.
39. Yang C, Lim W, Song G (2020). Mediation of oxidative stress toxicity induced by pyrethroid pesticides in fish. Gen Comp Endocrinol, 296, 113541.
40. Zhang X, Zhang T, Ren X et al. (2021). Pyrethroid toxicity to the male reproductive system and offspring mediated by oxidative stress: evidence from rodent studies. Front Endocrinol, 12, 656106.
41. Zheng W, Xu L, Jin M, Wang J, Rietjens IMCM (2024). Effects of lambda-cyhalothrin on gut microbiota and related bile acid metabolism in mice. J Hazard Mater, 480, 136417.
42. Zotti M, Colaianna M, Morgese MG et al. (2013). Carvacrol: from ancient flavoring to neuromodulatory agent. Molecules, 18, 6161–6172.