Abstract
SGLT2 inhibitors (SGLT2i) slow the progression of chronic kidney disease; however, evidence for the underlying molecular mechanisms is scarce. We investigated SGLT2i-mediated effects on differential gene expression in two independent human proximal tubular cell (HPTC) lines (HK-2 and RPTEC/TERT1) at the mRNA and protein levels under normoglycemic conditions, utilizing IL-1β as a pro-inflammatory mediator. Microarray hybridization identified 259 genes that were uniformly upregulated by IL-1β (10 mg/mL) and downregulated by empagliflozin (Empa) (500 nM) after 24 h of stimulation in two independent HPTC lines (n = 2, each). The functional annotation of these genes identified eight pathway clusters. Among 12 genes annotated to the highest ranked cluster (enrichment score, 3.51), monocyte chemoattractant protein-1/CC-chemokine ligand 2 (MCP-1/CCL2) and endothelin-1 (ET-1) were selected for verification at mRNA and protein levels based on their established involvement in the early pathogenesis of chronic kidney disease: IL-1β upregulated basal MCP-1/CCL2 (15- and 19-fold) and ET-1 (3- and 8-fold) mRNA expression, while Empa downregulated basal MCP-1/CCL2 (0.6- and 0.5-fold) and ET-1 (0.3- and 0.2-fold) mRNA expression as early as 1 h after stimulation and for at least 24 h in HK-2 and RPTEC/TERT1 cells, respectively. The co-administration of Empa inhibited IL-1β-mediated MCP-1/CCL2 (0.2-fold, each) and ET-1 (0.2-fold, each) mRNA expression as early as 1 h after ligand stimulation and for at least 24 h in both HPTC lines, respectively. This inhibitory effect of Empa on basal and IL-1β-mediated MCP-1/CCL2 and ET-1 mRNA expression was corroborated at the protein level. Our study presents novel evidence for the interference of SGLT2 inhibition with tubular inflammatory response mechanisms under normoglycemic conditions that might account for SGLT2i-mediated nephroprotection.
Highlights
Diabetic kidney disease (DKD) is a major cause of chronic kidney disease (CKD) and end-stage renal disease worldwide [1] and markedly influences both the quality and quantity of life [2,3,4]
We investigated SGLT2 inhibitors (SGLT2i)-mediated effects on differential gene expression in two independent human proximal tubular cell (HPTC) lines (HK-2 and RPTEC/TERT1) at the mRNA and protein levels under normoglycemic conditions, utilizing IL-1β as a pro-inflammatory mediator
Among 12 genes annotated to the highest ranked cluster, monocyte chemoattractant protein-1/CC-chemokine ligand 2 (MCP-1/CCL2) and endothelin-1 (ET-1) were selected for verification at mRNA and protein levels based on their established involvement in the early pathogenesis of chronic kidney disease: IL-1β upregulated basal MCP-1/CCL2 (15- and 19-fold) and ET-1 (3- and 8-fold) mRNA expression, while Empa downregulated basal MCP-1/CCL2 (0.6- and 0.5-fold) and ET-1 (0.3- and 0.2-fold) mRNA expression as early as 1 h after stimulation and for at least 24 h in Human kidney 2 (HK-2) and RPTEC/TERT1 cells, respectively
Summary
Diabetic kidney disease (DKD) is a major cause of chronic kidney disease (CKD) and end-stage renal disease worldwide [1] and markedly influences both the quality and quantity of life [2,3,4]. The DAPA-CKD trial recently demonstrated that dapagliflozin reduces the risk of eGFR decline, end-stage renal disease, or death from renal or CV causes in CKD patients regardless of the presence or absence of diabetes [20]. By demonstrating that SGLT2i inhibit the basal and TGF-β1-mediated expression of thrombospondin 1 (THBS1), tenascin-C (TNC) and platelet-derived growth factor-beta (PDGF-B) in two independent human proximal tubular cell (HPTC) lines under normoglycemic conditions, we recently presented non-hemodynamic, nephroprotective mechanisms of this promising class of drugs [22]. By demonstrating an empagliflozin (Empa)-mediated inhibition of basal as well as IL-1β-mediated MCP-1/CCL2 and ET-1 expression under normoglycemic conditions, we present novel evidence for the interference of SGLT2 inhibition with tubular inflammatory response mechanisms that could convey SGLT2i-mediated renoprotection in normoglycemic CKD patients
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