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

Articles in Press

Document Type : Original Article

Authors

1 Department of Biochemistry, Faculty of Veterinary Medicine, Ataturk University, Erzurum, Türkiye

2 Department of Histology-Embryology, Faculty of Medicine, Kafkas University, Kars, Türkiye

3 Horasan Vocational College, Ataturk University, Erzurum, Türkiye

4 Department of Medical Laboratory Techniques, Vocational School of Health Services, Atatürk University, Erzurum, Türkiye

5 Department of Pharmacy Services, Nihat Delibalta Göle Vocational High School, Ardahan University, Ardahan, 75700, Türkiye

6 Department of Nursing, Faculty of Health Sciences, Mardin Artuklu University, Mardin, Türkiye

10.22038/ijbms.2025.90102.19424

Abstract

Objective(s): Diazinon (DZN), a widely used organophosphate pesticide, induces pulmonary toxicity through oxidative stress, inflammation, endoplasmic reticulum (ER) stress, and apoptosis. This study investigated the potential protective effects of protocatechuic acid (PCA) against DZN-induced lung injury in rats.
Materials and Methods: Thirty-five adult rats were randomly assigned to five groups (n = 7): Control, DZN (20 mg/kg), PCA100 (100 mg/kg), DZN + PCA50, and DZN + PCA100. Lung tissues were evaluated histopathologically, and oxidative stress markers (GSH, SOD, CAT, and GPx) and inflammatory mediators (TNF-α, IL-1β, IL-6, NF-κB, COX-2, and iNOS) were measured by ELISA. The protein levels of Keap-1, Nrf2, and HO-1 were assessed via Western blotting. Expression of ER stress–related genes (XBP-1, eIF2, ATF4, CHOP) and apoptotic markers (Bax, Bcl-2, caspase-3, -6, -9) was analyzed by qRT-PCR.
Results: DZN exposure caused severe histopathological damage and significantly increased oxidative, inflammatory, ER stress, and apoptotic responses. PCA administration, particularly at 100 mg/kg, markedly improved lung morphology, normalized antioxidant enzyme levels, reduced cytokine production and NF-κB activation, and downregulated ER stress and apoptosis-related genes. PCA also enhanced Bcl-2 expression and activated the Nrf2/HO-1 signaling pathway.
Conclusion: PCA exerts dose-dependent protective effects against DZN-induced pulmonary toxicity by modulating oxidative stress, inflammation, ER stress, and apoptosis. These findings suggest that PCA may serve as a promising therapeutic candidate for mitigating pesticide-related lung injury.

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References

1. Zeinali M, Shafaei A, Rafatpanah H, Mosavat A, Tayebi-Meybodi N, Hosseinzadeh H, et al. Potential protective effects of chrysin against immunotoxicity induced by diazinon. Sci Rep 2022; 12:1–13. 
2. Safaeian L, Haghighatian Z, Zamani M. Cardioprotective potential of protocatechuic acid against doxorubicin-induced cardiotoxicity in rats. Res Pharm Sci 2025; 20:55–64. 
3. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 2001; 25:402–408. 
4. Bass JJ, Wilkinson DJ, Rankin D, Phillips BE, Szewczyk NJ, Smith K, et al. An overview of technical considerations for Western blotting applications to physiological research. Scand J Med Sci Sports 2017; 27:4–25. 
5. Dogan T, Yildirim BA, and Kapakin KAT. Investigation of the effects of crocin on inflammation, oxidative stress, apoptosis, NF-κB, TLR-4 and Nrf-2/HO-1 pathways in gentamicin-induced nephrotoxicity in rats. Environ Toxicol Pharmacol 2024; 106:104374. 
6. Wu X, Li J, Zhou Z, Lin Z, Pang S, Bhatt P, et al. Environmental occurrence, toxicity concerns, and degradation of diazinon using a microbial system. Front Microbiol 2021; 12:717286. 
7. Esfahani M and Mehri F. Homeostatic changes of trace elements in diazinon toxicity in rat model: The beneficial role of resveratrol. Toxicol Rep 2024; 13:101719. 
8. Hernández-Moreno D, Míguez MP, Soler F, Pérez-López M. Influence of sex on biomarkers of oxidative stress in the kidney, lungs, and liver of rabbits after exposure to diazinon. Environ Sci Pollut Res Int  2018; 25:32458–32465. 
9. Ivanović SR, Borozan N, Miladinović DĆ, Živković I, Borozan S. The relationship between the cholinergic mechanism of toxicity and oxidative stress in rats during subacute diazinon poisoning. Toxicol Appl Pharmacol 2023; 473:116598. 
10. Yu J. Inflammatory mechanisms in the lung. J Inflamm Res 2009; 2:1-11.
11. Niazi R, Peeri M, and Azarbayjani MA. The interactive effect of berberine chloride and exercise rehabilitation on the lung tissue apoptosis and oxidative stress biomarkers in rats exposed to diazinon. Physiol Pharmacol 2022; 26:60–69. 
12. Rady MI. Effects of exposure to Diazinon on the lung and small intestine of Guinea pig, histological and some histochemical changes. Braz Arch Biol Technol 2009; 52:317–326. 
13. Kelidari M, Abedi F, Hayes AW, Jomehzadeh V, and Karimi G. The protective effects of protocatechuic acid against natural and chemical toxicants: Cellular and molecular mechanisms. Naunyn Schmiedebergs Arch Pharmacol 2024; 397:5597–5616. 
14. Salama A, Elgohary R, Amin MM, and Elwahab SA. Immunomodulatory effect of protocatechuic acid on cyclophosphamide induced brain injury in rat: Modulation of inflammosomes NLRP3 and SIRT1. Eur J Pharmacol 2022; 932:175217. 
15. Abdelmageed ME, Nader MA, and Zaghloul MS. Targeting HMGB1/TLR4/NF-κB signaling pathway by protocatechuic acid protects against l-arginine induced acute pancreatitis and multiple organs injury in rats. Eur J Pharmacol 2021; 906:174279. 
16. Li L, Ma H, Zhang Y, Jiang H, Xia B, Sberi H Al, et al. Protocatechuic acid reverses myocardial infarction mediated by β-adrenergic agonist via regulation of Nrf2/HO-1 pathway, inflammatory, apoptotic, and fibrotic events. J Biochem Mol Toxicol 2023; 37:e23270. 
17. Yang MH, Baek SH, Chinnathambi A, Alharbi SA, Ahn KS. Identification of protocatechuic acid as a novel blocker of epithelial-to-mesenchymal transition in lung tumor cells. Phytother Res 2021; 35:1953–1966. 
18. Hashemi SS, Janfeshan S, and Karimi Z. Acute lung injury induced by acute uremia and renal ischemic-reperfusion injury: The role of toll-like receptors 2 and 4, and oxidative stress. Iran J Basic Med Sci 2022; 25:643–651. 
19. Karimi Z, Ketabchi F, Alebrahimdehkordi N, Fatemikia H, Owji SM, Moosavi SMS. Renal ischemia/reperfusion against nephrectomy for induction of acute lung injury in rats. Ren Fail 2016; 38:1503–1515. 
20. Abdel-Daim MM, Samak DH, El-Sayed YS, Aleya L, Alarifi S, Alkahtani S. Curcumin and quercetin synergistically attenuate subacute diazinon-induced inflammation and oxidative neurohepatic damage, and acetylcholinesterase inhibition in albino rats. Environ Sci Pollut Res Int 2019; 26:3659–3665. 
21. Kim J, Shin SH, Kang JK, and Kim JW. HX-1171 attenuates pancreatic β-cell apoptosis and hyperglycemia-mediated oxidative stress via Nrf2 activation in streptozotocin-induced diabetic model. Oncotarget 2018; 9:24260. 
22. Zhang X, Li C, Li J, Xu Y, Guan S, Zhao M. Protective effects of protocatechuic acid on acute lung injury induced by lipopolysaccharide in mice via p38MAPK and NF-κB signal pathways. Int Immunopharmacol 2015; 26:229–236. 
23. Anbarkeh FR, Nikravesh MR, Jalali M, Sadeghnia HR, Sargazi Z. The effect of diazinon on cell proliferation and apoptosis in testicular tissue of rats and the protective effect of vitamin E. Int J Fertil Steril 2019; 13:154-160. 
24. Sargazi Z, Nikravesh MR, Jalali M, Sadeghnia HR, Rahimi Anbarkeh F. Apoptotic effect of organophosphorus ınsecticide diazinon on rat ovary and protective effect of vitamin E. Iran J Toxicol 2016; 10:37–44. 
25. Razavi BM, Hosseinzadeh H, Movassaghi AR, Imenshahidi M, Abnous K. Protective effect of crocin on diazinon induced cardiotoxicity in rats in subchronic exposure. Chem Biol Interact 2013; 203:547–555. 
26. Karaca O, Şimşek H, Akaras N, Gür C, İleritürk M, Kandemir Ö, et al. Potent ameliorative effects of rosmarinic acid on tramadol-ınduced neurotoxicity in the brain and hippocampus; by suppressing oxidative stress, apoptosis, er stress, and regulating cognitive functions. Mol Neurobiol 2025; 62: 10245-10263.
27. Thakur S, Dhiman M, Mantha AK. APE1 modulates cellular responses to organophosphate pesticide-induced oxidative damage in non-small cell lung carcinoma A549 cells. Mol Cell Biochem 2018; 441:201–216. 
28. Boussabbeh M, Ben Salem I, Hamdi M, Ben Fradj S, Abid-Essefi S, Bacha H. Diazinon, an organophosphate pesticide, induces oxidative stress and genotoxicity in cells deriving from large intestine. Environ Sci Pollut Res Int 2016; 23:2882–2889. 
29. Lee WJ and Lee SH. Protocatechuic acid protects hepatocytes against hydrogen peroxide-induced oxidative stress. Curr Res Food Sci 2022; 5:222–227. 
 
Iranian Journal of Basic Medical Sciences

Articles in Press, Accepted Manuscript
Available Online from 28 December 2025

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