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Background/objectives: The porcine intestinal epithelium plays a fundamental role as a defence interface against pathogens. Its alteration can cause severe inflammatory conditions and diseases. Hyperosmotic stress under physiological conditions and upon pathogen challenge can cause malabsorption. Different cell types counteract the osmolarity increase by accumulating organic osmolytes such as betaine, taurine, and myo-inositol through specific transporters. Betaine is known for protecting cells from hyperosmotic stress and has positive effects when fed to pigs. The aim of this study is to demonstrate the modulation of osmolyte transporters gene expression in IPEC-J2 during osmolarity changes and assess the effects of betaine. Methods: IPEC-J2 were seeded in transwells, where differentiate as a polarized monolayer. Epithelial cell integrity (TEER), oxidative stress (NO) and gene expression of osmolyte transporters, tight junction proteins (TJp) and pro-inflammatory cytokines were evaluated. Results: Cells treated with NaCl hyperosmolar medium (500 mOsm/L) showed a TEER decrease at 3 h and detachment within 24 h, associated with an osmolyte transporters reduction. IPEC-J2 treated with mannitol hyperosmolar medium (500 mOsm/L) upregulated taurine (TauT), myo-inositol (SMIT) and betaine (BGT1) transporters expression. A decrease in TJp expression was associated with a TEER decrease and an increase in TNFα, IL6, and IL8. Betaine could attenuate the hyperosmolarity-induced reduction in TEER and TJp expression, the NO increase and cytokines upregulation. Conclusions: This study demonstrates the expression of osmolyte transporters in IPEC-J2, which was upregulated upon hyperosmotic treatment. Betaine counteracts changes in intracellular osmolarity by contributing to maintaining the epithelial barrier function and reducing the inflammatory condition. Compatible osmolytes may provide beneficial effects in therapies for diseases characterized by inflammation and TJp-related dysfunctions. [ABSTRACT FROM AUTHOR]

Bovine mastitis is among the most serious disease in the dairy industry and brings huge economic losses due to the decrease in milk quality and quantity. Betaine, a naturally occurring compound, possesses several pharmacological activities including anti-inflammatory and anti-oxidant ability, but whether betaine has protective effects on bovine mastitis is unknown. The aim of this study is to investigate the effect of betaine on mastitis and further discover its feasible molecular mechanism in lipopolysaccharide (LPS)-stimulated bovine mammary epithelial cells (BMECs). BMECs were pre-treated with or without betaine or LPS. Cell viability was measured with CCK-8 to examine the cytotoxicity. The levels of pro-inflammatory cytokines were measured by ELISA kits. Western blotting was used to explore the regulation of genes associated with inflammatory and oxidative stress genes. The results showed that LPS treatment significantly increased the production of pro-inflammation (IL-1β, IL-6 and TNFα), enhanced malondialdehyde (MDA) content, reduced superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity, and markedly up-regulated the protein expression of COX2 and iNOS (p < 0.05). However, betaine pre-treatment remarkably restored the above phenomenon compared with the LPS group. Additionally, we also observed that betaine exposure significantly restricted LPS-induced the phosphorylation of IκB and NF-κB p65 (p < 0.05). Moreover, pre-treatment of BMECs with betaine abolished LPS-induced the increase of Nrf2 and HO-1 protein levels (p < 0.05). These results confirm that betaine can alleviate LPS-induced inflammatory response and oxidative damage by modulating NF-κB and Nrf2 signalling pathways. Betaine alleviated the production of pro-inflammation cytokines (IL-1β, IL-6 and TNFα) in BMECs after LPS stimulation. Betaine restored LPS-elicited nuclear factor kappa-B (NF-κB) signalling pathway activity. Betaine recovered LPS-induced the activity of SOD and GSH-Px and MDA content. Betaine inhibited the increase of protein expression of Nrf2 and HO-1 challenged with LPS. [ABSTRACT FROM AUTHOR]