Related Experiment Video
Updated: Oct 9, 2026
Phenotypic Characterization of Macrophages from Rat Kidney by Flow Cytometry
Published on: October 18, 2016
Empagliflozin reduces kidney fibrosis and improves kidney function by alternative macrophage activation in rats with
Yong-Ping Lu1, Hong-Wei Wu1, Ting Zhu1
1Department of Nephrology, Center of Kidney and Urology, the Seventh Affiliated Hospital, Sun Yat-sen University, Shenzhen, China.
Background:
Sodium glucose cotransporter 2 (SGLT2) inhibitors originally developed for the treatment of type 2 diabetes are clinically very effective drugs halting chronic kidney disease progression. The underlying mechanisms are, however, not fully understood.
Methods:
We generated single-cell transcriptomes of kidneys from rats with 5/6 nephrectomy before and after SGLT2 inhibitors treatment by single-cell RNA sequencing.
Findings:
Empagliflozin treatment decreased BUN, creatinine and urinary albumin excretion compared to placebo by 39.8%, 34.1%, and 55%, respectively (p < 0.01 in all cases). Renal interstitial fibrosis and glomerulosclerosis was likewise decreased by 51% and 66.8%; respectively (p < 0.05 in all cases). 14 distinct kidney cell clusters could be identified by scRNA-seq. The polarization of M2 macrophages from state 1 (CD206-CD68- M2 macrophages) to state 5 (CD206+CD68+ M2 macrophages) was the main pro-fibrotic process, as CD206+CD68+ M2 macrophages highly expressed fibrosis-promoting genes and can convert into fibrocytes. Empagliflozin remarkably inhibited the expression of fibrosis-promoting (IFG1 and TREM2) and polarization-associated genes (GPNMB, LGALS3, PRDX5, and CTSB) in CD206+CD68+ M2 macrophages and attenuated inflammatory signals from CD8+ effector T cells. The inhibitory effect of empagliflozin on CD206+CD68+ M2 macrophages polarization was mainly achieved by affecting mitophagy and mTOR pathways.
Interpretation:
We propose that the beneficial effects of empagliflozin on kidney function and morphology in 5/6 nephrectomyiced rats with established CKD are at least partially due to an inhibition of CD206+CD68+ M2 macrophage polarization by targeting mTOR and mitophagy pathways and attenuating inflammatory signals from CD8+ effector T cells.
Fundings:
A full list of funding bodies that contributed to this study can be found in the Acknowledgements section.
Insights
Sodium glucose cotransporter 2 (SGLT2) inhibitors like empagliflozin halt chronic kidney disease progression by inhibiting M2 macrophage polarization. This mechanism targets mTOR and mitophagy pathways, reducing fibrosis and improving kidney function.
Area of Science:
- Nephrology
- Immunology
- Pharmacology
Background:
- Sodium glucose cotransporter 2 (SGLT2) inhibitors are effective in slowing chronic kidney disease (CKD) progression.
- The precise mechanisms underlying their renoprotective effects remain incompletely understood.
Purpose of the Study:
- To investigate the molecular mechanisms by which SGLT2 inhibitors exert their beneficial effects on kidney disease.
- To analyze the impact of empagliflozin on cellular and molecular changes in a rat model of CKD.
Main Methods:
- Single-cell RNA sequencing was employed to analyze kidney tissue from rats with 5/6 nephrectomy before and after empagliflozin treatment.
- Key cellular populations and gene expression patterns associated with fibrosis and inflammation were identified.
Main Results:
- Empagliflozin significantly reduced markers of kidney injury (BUN, creatinine, albuminuria) and histological damage (fibrosis, glomerulosclerosis).
- Single-cell analysis revealed that empagliflozin inhibited the polarization of pro-fibrotic M2 macrophages (CD206+CD68+) by targeting mitophagy and mTOR pathways.
- Empagliflozin also attenuated inflammatory signals from CD8+ effector T cells.
Conclusions:
- The renoprotective effects of empagliflozin in CKD are, at least partially, mediated by the inhibition of M2 macrophage polarization.
- Targeting mTOR and mitophagy pathways, along with reducing CD8+ T cell inflammation, contributes to empagliflozin's efficacy in established kidney disease.
