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Int J Mol Sci ; 21(21)2020 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-33139635

RESUMO

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-1ß as a pro-inflammatory mediator. Microarray hybridization identified 259 genes that were uniformly upregulated by IL-1ß (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-1ß 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-1ß-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-1ß-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.


Assuntos
Compostos Benzidrílicos/farmacologia , Quimiocina CCL2/genética , Endotelina-1/genética , Expressão Gênica/efeitos dos fármacos , Glucosídeos/farmacologia , Interleucina-1beta/farmacologia , Túbulos Renais Proximais/efeitos dos fármacos , Linhagem Celular , Quimiocina CCL2/metabolismo , Endotelina-1/metabolismo , Perfilação da Expressão Gênica/métodos , Humanos , Túbulos Renais Proximais/citologia , Túbulos Renais Proximais/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos/métodos
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