Sex-specific modulation of gut microbiota by empagliflozin contributes to renoprotection in diabetic kidney disease mice

Objectives This study aimed to explore the sex-specific regulatory effects of empagliflozin on gut microbiota in male and female diabetic kidney disease (DKD) mice, and to elucidate the underlying renoprotective mechanisms. Methods Four-week-old db/db mice and C57 mice were randomly assigned to six groups. Following 8 weeks of empagliflozin gavage, serum metabolic indices and urinary albumin-to-creatinine ratio (ACR) were measured. Renal pathological alterations were evaluated via hematoxylin-eosin and Masson’s trichrome staining. Gut microbiota diversity and community composition were analyzed using 16S rRNA gene sequencing, and Spearman’s rank correlation analysis was performed to assess associations between dominant microbial taxa and metabolic/renal parameters. Results ACR levels were significantly elevated in db/db mice compared to sex-matched C57 controls, with male db/db mice exhibiting significantly higher ACR than females. Empagliflozin significantly reduced ACR in db/db mice, albeit ACR remained significantly higher in males than females post-treatment. Meanwhile, empagliflozin ameliorated glomerular hypertrophy and mesangial proliferation. Pronounced sexual differences were observed in gut microbiota diversity of db/db mice, with female mice displaying significantly higher microbial richness than males. Empagliflozin effectively reshaped gut microbiota composition and alleviated microbial dysbiosis in db/db mice, with these regulatory effects showing distinct sex specificity. Furthermore, several dominant microbial taxa (e.g., Rikenellaceae_RC9_gut_group , Parabacteroides , Klebsiella ) were identified to be significantly correlated with ACR, and these correlations were sex-dependent. Conclusion Empagliflozin significantly reduces ACR levels and modulates gut microbial richness and diversity in db/db mice, with distinct sex-specific effects on microbiota composition. Ultimately, these findings suggest that empagliflozin may exert renoprotective effects in DKD by reshaping gut microbial community structure.