Unraveling reno-protective effects of SGLT2 inhibition in human proximal tubular cells.

Large clinical trials demonstrated that SGLT2 inhibitors (SGLT2i) slow the progression of kidney function decline in type 2 diabetes. Because the underlying molecular mechanisms are largely unknown, we studied the effects of SGLT2i on gene expression in two human proximal tubular (PT) cell lines under normoglycemic conditions, utilizing two SGLT2i, namely empagliflocin and canagliflocin. Genome-wide expression analysis did not reveal substantial differences between these two SGLT2i. Microarray hybridization analysis identified 94 genes that were both upregulated by TGF-ß1 and downregulated by either of the two SGLT2i in HK-2 and RPTEC/TERT1 (renal proximal tubular epithelial cells/telomerase reverse transcriptase 1) cells. Extracellular matrix organization showed the highest significance in pathway enrichment analysis. Differential gene expression of three annotated genes of interest within this pathway was verified on mRNA level in both cell lines. Whereas TGF-ß1 induced mRNA expression of thrombospondin 1 (THBS1; 4.3-fold), tenascin C (TNC; 8-fold), and platelet-derived growth factor subunit B (PDGF-B; 4.2-fold), SGLT2i downregulated basal mRNA expression of THBS1 (0.2-fold), TNC (0.5 fold), and PDGF-B (0.6-fold). Administration of SGLT2i in the presence of TGF-ß1 resulted in a significant inhibition of TGF-ß1-induced THBS1 and TNC mRNA expression and TGF-ß1-induced THBS1, TNC, and PDGF-BB protein expression. We conclude that SGLT2i block basal and TGF-ß1-induced expression of key mediators of renal fibrosis and kidney disease progression in two independent human PT cell lines.

Acute calcium kinetics in haemodialysis patients.

INTRODUCTION: To avoid excessive calcium loading in haemodialysis (HD) patients, current guidelines suggest a dialysate calcium concentration (dCa) of 2·5 mEq/L based on relatively stable intradialytic serum calcium levels. However, the latter do not account for possible calcium storage in acutely accessible pools. A rapidly exchangeable calcium pool located at the bone level has been previously proposed to be involved in acute (minute-to-minute) extracellular calcium regulation. DESIGN: To evaluate the contribution of this pool in the maintenance of serum calcium levels, acute calcium buffer capacity was assessed by measuring intradialytic dialysate-sided ionized calcium mass balance (iCaMB ) and change in extracellular fluid calcium mass in chronic HD patients using a dCa of 3·5 (n = 28) and 2·5 (n = 10) mEq/L. Serum osteocalcin, the most abundant noncollagenous bone protein, was measured before the HD session. RESULTS: iCaMB was invariably positive for both 2·5 and 3·5 mEq/L dCa, with a mean of 434 (±125) and 725 (±162) mg/HD, respectively (P < 0·001). Buffered intradialytic calcium load was 410 (±116) and 565 (±130) mg/HD, and acute calcium buffer capacity was 95 (±8)% and 78 (±7)% (mean values at 2·5 and 3·5 mEq/L dCa, respectively) (P < 0·001). Using 3·5 mEq/L dCa, an independent association of acute calcium buffer capacity with undercarboxylated osteocalcin (ß = 0·512, P = 0·002) was demonstrated. CONCLUSIONS: Our data strongly suggest the existence of a rapidly exchangeable calcium pool that counteracts acute serum calcium deviations in HD patients. This study provides, for the first time, experimental evidence for the involvement of bone in acute extracellular calcium regulation in vivo.