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Effects of Cannabidiol on the In Vitro Lymphocyte Pro-Inflammatory Cytokine Production of Senior Horses.
Abstract: Cannabis sativa L. contains cannabidiol (CBD), a compound that has many anti-inflammatory properties. In this study, 99.9% CBD powder was used to determine its in vitro efficacy as an anti-inflammatory agent. Heparinized blood was collected via jugular venipuncture from senior horses. PBMCs were isolated then incubated for 24 hours with increasing dilutions of CBD dissolved in DMSO. PBMCs were stimulated the last 4 hours of incubation with PMA/IO and Brefeldin A. A Vicell counter was used to evaluate viability after incubation. PBMCs were stained intracellularly for IFNγ and TNFα then analyzed via flow cytometry. RT-PCR was used to analyze samples for gene expression. Five equine-specific intron-spanning primers/probes used are: CB1, CB2, TNFα, IFNγ, IL-10, and Beta-glucuronidase. Data was analyzed using RM One-way ANOVA (significance P < .05). Viability of PBMCs with CBD was completed to determine cytotoxicity. The dilution of CBD that did not affect cell viability was 4 µg/mL (P<0.05). CBD at 4 µg/mL significantly reduced production of IFN-γ and TNF-α (P < .05). RT-PCR results for TNFα and IFNγ at 4 µg/mL showed a reduction compared with the positive control and IL-10 showed a similar reduction at 2 µg/mL and 4 µg/mL. RT-PCR gene expression results showed significance for 10 μg/mL CBD in CB1 and CB2. CBD at 4 µg/mL reduced in vitro production of inflammatory cytokines from senior horses. This in vitro study supports further investigation of CBD to determine if it may be effective as an anti-inflammatory treatment for chronic inflammation in the horse.
Copyright © 2021. Published by Elsevier Inc.
Publication Date: 2021-05-24 PubMed ID: 34281647DOI: 10.1016/j.jevs.2021.103668Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The study examines whether Cannabidiol (CBD), a compound found in Cannabis sativa L., can inhibit the production of pro-inflammatory cytokines in elderly horses, which could signal its potential use as an anti-inflammatory treatment.
Methodology and Procedures
- The researchers used a 99.9% CBD powder for the study to test its efficacy as an anti-inflammatory agent.
- Heparinized blood was collected from elderly horses through jugular venipuncture.
- Peripheral Blood Mononuclear Cells (PBMCs) were extracted and then incubated for 24 hours with gradually increasing concentrations of the CBD solution.
- In the final four hours of incubation, the cells were stimulated with PMA/IO and Brefeldin A.
- A Vicell counter was employed to check the viability of the cells following the incubation period.
- Flow cytometry was subsequently used to analyze the PBMCs that had been internally stained for IFNγ and TNFα, two types of inflammatory cytokines.
- Gene expression of the samples was examined using RT-PCR, specifically focusing on five equine-specific primers — CB1, CB2, TNFα, IFNγ, IL-10, as well as Beta-glucuronidase.
Results of the Study
- The study revealed that a dilution of 4 µg/mL of CBD did not affect the viability of the PBMCs, indicating it wasn’t cytotoxic at this dosage.
- Furthermore, CBD at this concentration notably reduced the production of IFN-γ and TNF-α, both of which are pro-inflammatory cytokines.
- The RT-PCR analysis for TNFα and IFNγ at the same concentration revealed a similar decrease, while IL-10 showed a similar downturn at both 2 µg/mL and 4 µg/mL.
- Significant changes in gene expression were seen for 10 μg/mL CBD in CB1 and CB2.
Significance and Implications
- The findings suggest that CBD can reduce the in vitro production of inflammatory cytokines for elderly horses.
- This supports the premise that CBD could potentially be utilized as an anti-inflammatory treatment for chronic inflammation in horses.
- However, further investigations are necessary to confirm these findings and to explore if they apply in vivo (in the organism).
Cite This Article
APA
Turner S, Barker VD, Adams AA.
(2021).
Effects of Cannabidiol on the In Vitro Lymphocyte Pro-Inflammatory Cytokine Production of Senior Horses.
J Equine Vet Sci, 103, 103668.
https://doi.org/10.1016/j.jevs.2021.103668 Publication
Researcher Affiliations
- Department of Veterinary Science, M. H. Gluck Equine Research Center, University of Kentucky, Lexington, KY. Electronic address: Shelley.turner@uky.edu.
- Department of Veterinary Science, M. H. Gluck Equine Research Center, University of Kentucky, Lexington, KY.
- MARS Equestrian™ Research Fellow, Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington, KY, USA.
MeSH Terms
- Animals
- Anti-Inflammatory Agents / pharmacology
- Cannabidiol / pharmacology
- Cytokines / genetics
- Gene Expression
- Horses
- Lymphocytes
Citations
This article has been cited 9 times.- Leise JM, Leatherwood JL, Paris BL, Walter KW, George JM, Martinez RE, Glass KP, Lo CP, Mays TP, Wickersham TA. Evaluation of an Oral Supplemental Cannabidiol Product for Acceptability and Performance in Mature Horses. Animals (Basel) 2023 Jan 10;13(2).
- Filipiuc LE, Ababei DC, Alexa-Stratulat T, Pricope CV, Bild V, Stefanescu R, Stanciu GD, Tamba BI. Major Phytocannabinoids and Their Related Compounds: Should We Only Search for Drugs That Act on Cannabinoid Receptors?. Pharmaceutics 2021 Nov 1;13(11).
- da Costa Kamura B, de Oliveira Ferreira LV, Chimenes ND, de Oliveira JPM, Rodriguez-Sanchez DN, de Carvalho M, Amorim RM. Cannabidiol-rich extract suppresses the activation of proinflammatory genes IL-1β and IL-6 in equine mesenchymal stem cells stimulated with lipopolysaccharide. Vet Res Commun 2026 Feb 24;50(3).
- Zamith Cunha R, Gobbo F, Morini M, Salamanca G, Zanoni A, Bernardini C, Gramenzi A, Chiocchetti R. Cannabinoid and cannabinoid related receptors in fibroblasts, inflammatory and endothelial cells of the equine hoof with and without laminitis: novel pharmacological target. Front Vet Sci 2025;12:1723160.
- Battistin L, Moya LFA, Ferreira LVO, Braz AMM, Carvalho M, Golim MA, Amorim RM. In Vitro Immunomodulatory Effects of Equine Adipose Tissue-Derived Mesenchymal Stem Cells Primed with a Cannabidiol-Rich Extract. Int J Mol Sci 2025 Apr 29;26(9).
- Kosukwatthana P, Rungsuriyawiboon O, Rattanasrisomporn J, Kimram K, Tansakul N. Cytotoxicity and Immunomodulatory Effects of Cannabidiol on Canine PBMCs: A Study in LPS-Stimulated and Epileptic Dogs. Animals (Basel) 2024 Dec 20;14(24).
- Aragona F, Tabbì M, Gugliandolo E, Giannetto C, D'Angelo F, Fazio F, Interlandi C. Role of cannabidiolic acid or the combination of cannabigerol/cannabidiol in pain modulation and welfare improvement in horses with chronic osteoarthritis. Front Vet Sci 2024;11:1496473.
- Lyons C, McEwan K, Munn-Patterson M, Vuong S, Alcorn J, Chicoine A. Pharmacokinetic of two oral doses of a 1:20 THC:CBD cannabis herbal extract in cats. Front Vet Sci 2024;11:1352495.
- Interlandi C, Tabbì M, Di Pietro S, D'Angelo F, Costa GL, Arfuso F, Giudice E, Licata P, Macrì D, Crupi R, Gugliandolo E. Improved quality of life and pain relief in mature horses with osteoarthritis after oral transmucosal cannabidiol oil administration as part of an analgesic regimen. Front Vet Sci 2024;11:1341396.
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