Abstract

Microsporidia are obligate intracellular parasites associated with gastrointestinal disorders, including inflammatory bowel disease (IBD). Short-chain fatty acids (SCFAs), key microbial metabolites, play a crucial role in maintaining gut epithelial integrity and modulating immune responses. This study examines the effects of SCFAs on Microsporidia-infected CACO-2 cells grown on transwell filters, focusing on infection severity, barrier function, and cell viability.

CACO-2 cells were cultured on transwell inserts to establish a differentiated intestinal epithelial monolayer. Infection severity was assessed by collecting media from the apical and basolateral compartments and spore quantification using a hemocytometer. Transepithelial electrical resistance (TEER) was measured to evaluate barrier integrity, while paracellular permeability was assessed using fluorescent tracer molecules. Cell viability was determined to assess cytotoxic effects of infection and SCFA treatment.

Our results indicate that SCFAs significantly mitigate the impact of microsporidia infection on CACO-2 cells. SCFA-treated cells exhibited higher TEER values, indicating improved barrier integrity, and reduced paracellular permeability, suggesting enhanced tight junction function. Additionally, SCFAs increased cell viability and decreased spore burden compared to untreated infected controls. These findings suggest that SCFAs may play a protective role in reducing Microsporidia-induced epithelial damage, potentially influencing disease progression in IBD patients. Further research is needed to elucidate the molecular mechanisms by which SCFAs modulate host-pathogen interactions and to explore their therapeutic potential in microsporidia-associated infections.