The immunomodulatory effect of oral NaHCO 3 is mediated by the splenic nerve: multivariate impact revealed by artificial neural networks.

Stimulation of the inflammatory reflex (IR) is a promising strategy for treating systemic inflammatory disorders. Recent studies suggest oral sodium bicarbonate (NaHCO 3 ) as a potential activator of the IR, offering a safe and cost-effective treatment approach. However, the mechanisms underlying NaHCO 3 -induced anti-inflammatory effects remain unclear. We investigated whether oral NaHCO 3 's immunomodulatory effects are mediated by the splenic nerve. Female rats received NaHCO 3 or water (H 2 O) for four days, and splenic immune markers were assessed using flow cytometry. NaHCO 3 led to a significant increase (p < 0.05, and/or partial eta squared > 0.06) in anti-inflammatory markers, including CD11bc + CD206 + (M2-like) macrophages, CD3 + CD4 + FoxP3 + cells (Tregs), and Tregs/M1-like ratio. Conversely, proinflammatory markers, such as CD11bc + CD38 + TNFα + (M1-like) macrophages, M1-like/M2-like ratio, and SSC high /SSC low ratio of FSC high CD11bc + cells, decreased in the spleen following NaHCO 3 administration. These effects were abolished in spleen-denervated rats, suggesting the necessity of the splenic nerve in mediating NaHCO 3 -induced immunomodulation. Artificial neural networks accurately classified NaHCO 3 and H 2 O treatment in sham rats but failed in spleen-denervated rats, highlighting the splenic nerve's critical role. Additionally, spleen denervation independently influenced Tregs, M2-like macrophages, Tregs/M1-like ratio, and CD11bc + CD38 + cells, indicating distinct effects from both surgery and treatment. Principal component analysis (PCA) further supported the separate effects. Our findings suggest that the splenic nerve transmits oral NaHCO 3 -induced immunomodulatory changes to the spleen, emphasizing NaHCO 3 's potential as an IR activator with therapeutic implications for a wide spectrum of systemic inflammatory conditions.