The effects of N-acetyl tryptophan and omega-3 feeding during the transition period on relative expression genes related to endocannabinoid system in adipose tissue of Holstein cows

Articles in Press

Document Type : Original Articles

Authors

1 Department of Animal Science, Sari Agricultural Science and Natural Resources University, Sari, Iran.

2 Department of Clinical Science, Faculty of Veterinary Medicine, University of Tabriz, 5166616471 Tabriz, Iran

3 University of Florida

10.22092/ari.2025.369612.3682

Abstract

The transition period in dairy cows is marked by significant metabolic, endocrine, and immunological transformations, often contributing to negative energy balance and enhanced susceptibility to metabolic disease. The endocannabinoid system (ECS) contributes to regulating energy homeostasis and inflammation and its activation during this period has been implicated in insulin resistance, and reproductive dysfunctions. This research investigated the impact of dietary supplementation with omega-3 fatty acids (100 gr/cow/day) and tryptophan (TRP) amino acid (100 gr/cow/day), and interaction if these components, upon ECS-related gene expression in Holstein dairy cow adipose tissue at day 21 and 42 postpartum. Cows underwent presynchronization with two injections of prostaglandin F2α (PGF2α) administered 14 days apart. Following the second PGF2α injection, an Ovsynch protocol started with a GnRH injection, all cows artificially inseminated, and pregnancy accessed via ultrasonography on day 33 post-insemination. A designated area of 25 cm² (5×5 cm) was shaved and disinfected scalpel was used to collect of the subcutaneous adipose tissue (AT) sample. The obtained sample rinsed with distilled water, and placed into a microtube and stored at -80°C for measuring the relative expression of genes related to ECS: cannabinoid receptor 1 (CNR1), cannabinoid receptor 2 (CNR2), fatty acid amide hydrolase (FAAH), monoglyceride lipase (MGLL), N-acyl phosphatidylethanolamine phospholipase D (NAPEPLD), N-acylethanolamine acid amidase (NAAA), Cyclooxygenase 2 (COX2), and N-arachidonoyl phosphatidylethanolamine (NAPEPLD). The findings showed that interaction of omega-3 and TRP supplementation upregulated the relative expression of ECS-related genes, such as NAAA, FAAH, MGLL, and downregulated the expression of CNR2 and NAPEPLD, with no significant effects of CNR1 and COX2 on day 21 and 42 of postpartum. These results demonstrate a synergistic effect between omega-3 and TRP in modulating ECS activity, suppressing inflammation, and enhancing lipid metabolism. Downregulation of lipogenic and pro-inflammatory genes found in these results supports that targeted nutritional changes can modulate ECS activity, enhance negative energy balance, and increase metabolic conditions in transition dairy cows.

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Archives of Razi Institute

Articles in Press, Accepted Manuscript
Available Online from 09 August 2025

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