Cannabidiol attenuates lung ischemia-reperfusion injury by modulating RIPK1/RIPK3-mediated necroptosis and HIF-1α/VEGF signaling

Articles in Press

Document Type : Original Article

Authors

1 Department of Thoracic Surgery, Faculty of Medicine, Suleyman Demirel University, Isparta, Türkiye

2 Department of Pharmacology, Faculty of Medicine, Suleyman Demirel University, Isparta 32200, Türkiye

3 Department of Biochemistry, Faculty of Medicine, Suleyman Demirel University, Isparta 32200, Türkiye

4 Department of Genetic, Faculty of Medicine, Suleyman Demirel University, Isparta 32200, Türkiye

5 Department of Pathology, Faculty of Veterinary, Burdur Mehmet Akif Ersoy University, Burdur 15000, Türkiye

10.22038/ijbms.2026.90024.19406

Abstract

Objective(s): Lung ischemia-reperfusion (IR) injury is a critical clinical condition characterized by oxidative stress, inflammation, and necroptosis, often leading to severe complications. Cannabidiol (CBD), a non-psychoactive cannabinoid, has demonstrated anti-oxidant and anti-inflammatory properties, but its role in modulating lung IR injury remains incompletely understood. This study investigated the protective effects of CBD on lung IR injury in rats, focusing on the RIPK1/RIPK3 necroptosis pathway and the HIF-1α/VEGF/eNOS signaling axis.
Materials and Methods: Forty male Wistar albino rats were randomized into four groups: control, IR, IR+CBD (5 mg/kg), and CBD-only. Histopathological, immunohistochemical (TNF-α, Caspase-3), biochemical (TOS, TAS, OSI), and gene expression (RIPK1, RIPK3, HIF-1α, VEGF, eNOS) analyses were performed. The IR group exhibited significant oxidative stress, inflammation, and tissue damage, with elevated TNF-α, Caspase-3, TOS, OSI, and necroptosis/apoptosis markers.
Results: CBD treatment markedly attenuated these effects, reducing oxidative stress (↑TAS, ↓TOS/OSI), suppressing inflammation (↓TNF-α), and inhibiting both apoptotic (↓Caspase-3) and necroptotic (↓RIPK1/RIPK3) pathways. Additionally, CBD down-regulated HIF-1α/VEGF/eNOS expression, suggesting modulation of hypoxia-responsive signaling. 
Conclusion: These findings demonstrate that CBD mitigates lung IR injury by targeting oxidative stress, inflammation, and cell death mechanisms, highlighting its potential as a therapeutic agent. Further preclinical and clinical studies are warranted to validate these results.

Keywords

Main Subjects

References

1. Kandilci HB, Gümüşel B. Akciğerlerde i̇skemi-reperfüzyon hasarı ve i̇skemik önkoşullama. Hacettepe Univ J Fac Pharm 2005; 1:35–49. 
2. Alaçam B, Ek RO, Yıldız Y, Serter M, Boylu NT, Temoçın S. The effect of carnosine on lung injury induced by abdominal aortic ischemia-reperfusion. Meandros Med Dent J 2010;11:41–47. 
3. Tang H, Sun L, Huang J, Yang Z, Li C, Zhou X. The mechanism and biomarker function of Cavin-2 in lung ischemia-reperfusion injury. Comput Biol Med 2022;151:106234. 
4.    Kim EK, Jang M, Song MJ, Kim D, Kim Y, Jang HH. Redox-mediated mechanism of chemoresistance in cancer cells. Antioxidants 2019;8:471. 
5.    Atalay S, Jarocka-Karpowicz I, Skrzydlewska E. Antioxidative and anti-inflammatory properties of cannabidiol. Antioxidants 2019;9:21. 
6.    Zhang Z, Yao L, Yang J, Wang Z, Du G. PI3K/Akt and HIF‑1 signaling pathway in hypoxia‑ischemia. Mol Med Rep 2018;18:3547–3554. 
7. Khosropoor S, Alavi MS, Etemad L, Roohbakhsh A. Cannabidiol goes nuclear: The role of PPARγ. Phytomedicine 2023;114:154771.
8.    Liu C, Pei S, Dai H, Liu Z, Ye M, Liu H, et al. Downregulation of SIRT3 aggravates lung ischemia reperfusion injury by increasing mitochondrial fission and oxidative stress through HIF‐1α‐Dependent mechanisms. Oxid Med Cell Longev 2022;2022:9041914. 
9.    Zhou J, Schmid T, Brune B. Tumor necrosis factor-α causes accumulation of a ubiquitinated form of hypoxia inducible factor-1α through a nuclear factor-κB-dependent pathway. Mol Biol Cell 2003;14:2216–2225. 
10.    Syukri A, Hatta M, Amir M, Rohman MS, Mappangara I,  Kaelan C, et al. Doxorubicin induced immune abnormalities  and inflammatory responses via HMGB1, HIF1-α and VEGF pathway in progressive of cardiovascular damage.  Ann Med Surg 2022;76:103501.
11.    Qiu Y, Chen C, Zhang J, Chen M, Gong H, Gong L, et al. VEGF attenuates lung injury by inducing homing of CD133+ progenitors via VEGFR1. Biochem Biophys Res Commun 2019;511:650–657. 
12.    Savitskaya MA, Onishchenko GE. Mechanisms of apoptosis. Biochem 2015;80:1393–1405. 
13.    Coşkun G, Özgür H. Apoptoz ve nekrozun moleküler mekanizması. Arşiv Kaynak Tarama Derg 2011;20:145–158. 
14.    Wani AK, Akhtar N, Mir T ul G, Singh R, Jha PK, Mallik SK, et al. Targeting apoptotic pathway of cancer cells with phytochemicals and plant-based nanomaterials. Biomolecules 2023;13:194. 
15.    Obeng E. Apoptosis (programmed cell death) and its signals-A review. Brazilian J Biol 2020;81:1133–1143. 
16.    Delhalle S, Duvoix A, Schnekenburger M, Morceau F, Dicato M, Diederich M. An introduction to the molecular mechanisms of apoptosis. Ann N Y Acad Sci 2003;1010:1–8. 
17.    Mizumura K, Maruoka S, Gon Y, Choi AMK, Hashimoto S. The role of necroptosis in pulmonary diseases. Respir Investig 2016;54:407–412. 
18.    Shan B, Pan H, Najafov A, Yuan J. Necroptosis in development and diseases. Genes Dev. 2018;32:327–340. 
19.    Jun W, Benjanuwattra J, Chattipakorn SC, Chattipakorn N. Necroptosis in renal ischemia/reperfusion injury: a major mode of cell death? Arch Biochem Biophys 2020;689:108433. 
20.    Yu Z, Jiang N, Su W, Zhuo Y. Necroptosis: a novel pathway in neuroinflammation. Front Pharmacol 2021;12:701564. 
21.    Arruza L, Pazos MR, Mohammed N, Escribano N, Lafuente H, Santos M, et al. Cannabidiol reduces lung injury induced by hypoxic–ischemic brain damage in newborn piglets. Pediatr Res 2017;82:79–86. 
22.    Erdem S, Gonca E, Başoğlu G, Aydemir E. İskemi Sonrası Verilen Kannabidiol Reperfüzyon ile Uyarılan Aritmilere Karşı Koruyucu Etkilidir. Batı Karadeniz Tıp Derg 2022;6:16–23. 
23.    Kicman A, Toczek M. The effects of cannabidiol, a non-intoxicating compound of cannabis, on the cardiovascular system in health and disease. Int J Mol Sci 2020;21:6740. 
24.    Etemad L, Karimi G, Alavi MS, Roohbakhsh A. Pharmacological effects of cannabidiol by  transient receptor potential channels. Life Sci 2022;300:120582.
25.    Pereira SR, Hackett B, O’Driscoll DN, Sun MC, Downer EJ. Cannabidiol modulation of oxidative stress and signalling. Neuronal Signal 2021;5:NS20200080. 
26.    Abu‐Amara M, Yang SY, Tapuria N, Fuller B, Davidson B, Seifalian A. Liver ischemia/reperfusion injury: Processes in inflammatory networks—a review. Liver Transplant 2010;16:1016–1032. 
27.    Akin SE, Asci H, Tepebasi MY, Ilhan İ, Ozmen Ö, Comlekci S, et al. Prophylactic effects of radiofrequency electromagnetic field on pulmonary ischemia‐reperfusion via HIF‐1α/eNOS pathway and BCL2/BAX signaling. Pulm Circ 2025;15:e70051. 
28.    İlhan İ, Ascı H, Buyukbayram Hİ, Imeci OB, Sevuk MA,  Erol Z, et al. The impact of the high-fructose corn syrup  on cardiac damage via SIRT1/PGC1-α pathway: Potential  ameliorative effect of selenium. Biol Trace Elem Res 2024;202: 5166-5176.
29.    Tveden-nyborg P, Lykkesfeldt J. BCPT 2023 policy for experimental and clinical studies. Basic Clin Pharmacol Toxicol 2023;133:391–396. 
30.    Ozmen O, Milletsever A, Tasan S, Selcuk E, Savran M. The effects of cannabidiol against Methotrexate-induced lung damage. Basic Clin Pharmacol Toxicol 2024;134:695–703. 
31.    Topsakal S, Ozmen O, Asci H, Gulal A, Ozcan KN, Aydin B. Dapagliflozin prevents reproductive damage caused by acute systemic inflammation through antioxidant, anti‐inflammatory, and antiapoptotic mechanisms. Basic Clin Pharmacol Toxicol 2024;135:561–574. 
32.    Halladin NL. Oxidative and inflammatory biomarkers of ischemia and reperfusion injuries. Dan Med J 2015;62:B5054. 
33.    Akkoç H, Kelle İ, Kale E. Sıçan akciğerinde oluşturulan iskemik önkoşullamanın kalpteki iskemi reperfüzyon hasarı üzerine etkileri. Dicle Tıp Derg 2008;35:102–109. 
34.    Solinas M, Massi P, Cinquina V, Valenti M, Bolognini D, Gariboldi M, et al. Cannabidiol, a non-psychoactive cannabinoid compound, inhibits proliferation and invasion in U87-MG and T98G glioma cells through a multitarget effect. PLoS One 2013;8:e76918. 
35. He H, Venema VJ, Gu X, Venema RC, Marrero MB, Caldwell RB. Vascular endothelial growth factor signals endothelial cell production of nitric oxide and prostacyclin through flk-1/KDR activation of c-Src. J Biol Chem 1999;274:25130–25135.
36. Fukumura D, Gohongi T, Kadambi A, Izumi Y, Ang J, Yun CO, et al. Predominant role of endothelial nitric oxide synthase in vascular endothelial growth factor-induced angiogenesis and vascular permeability. Proc Natl Acad Sci 2001;98:2604–2609.
37.    Esposito G, De Filippis D, Maiuri MC, De Stefano D, Carnuccio R, Iuvone T. Cannabidiol inhibits inducible nitric oxide synthase protein expression and nitric oxide production in β-amyloid stimulated PC12 neurons through p38 MAP kinase and NF-κB involvement. Neurosci Lett 2006;399:91–95. 
38.    Lundberg JO, Weitzberg E. Nitric oxide signaling in health and disease. Cell 2022;185:2853–2878. 
39.    Khaksar S, Bigdeli M, Samiee A, Shirazi-Zand Z. Antioxidant and anti-apoptotic effects of cannabidiol in model of ischemic stroke in rats. Brain Res Bull 2022;180:118–130. 
40.    Mukhopadhyay P, Rajesh M, Horváth B, Bátkai S, Park O, Tanchian G, et al. Cannabidiol protects against hepatic ischemia/reperfusion injury by attenuating inflammatory signaling and response, oxidative/nitrative stress, and cell death. Free Radic Biol Med 2011;50:1368–1381. 
41.    Corpetti C, Del Re A, Seguella L, Palenca I, Rurgo S, De Conno B, et al. Cannabidiol inhibits SARS‐Cov‐2 spike (S) protein‐induced cytotoxicity and inflammation through a PPARγ‐dependent TLR4/NLRP3/Caspase‐1 signaling suppression in Caco‐2 cell line. Phyther Res 2021;35:6893–6903. 
42.    Fouad AA, Al-Mulhim AS, Jresat I. Cannabidiol treatment ameliorates ischemia/reperfusion renal injury in rats. Life Sci 2012;91:284–292. 
Iranian Journal of Basic Medical Sciences

Articles in Press, Accepted Manuscript
Available Online from 21 April 2026

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