RESUMO
This work was designed to further study the mechanism by which sulforaphane (SFN) exerts a renoprotective effect against cisplatin (CIS)-induced damage. It was evaluated whether SFN attenuates the CIS-induced mitochondrial alterations and the impairment in the activity of the cytoprotective enzymes NAD(P)H: quinone oxidoreductase 1 (NQO1) and γ glutamyl cysteine ligase (γGCL). Studies were performed in renal epithelial LLC-PK1 cells and in isolated renal mitochondria from CIS, SFN or CIS+SFN treated rats. SFN effectively prevented the CIS-induced increase in reactive oxygen species (ROS) production and the decrease in NQO1 and γGCL activities and in glutathione (GSH) content. The protective effect of SFN on ROS production and cell viability was prevented by buthionine sulfoximine (BSO), an inhibitor of γGCL, and by dicoumarol, an inhibitor of NQO1. SFN was also able to prevent the CIS-induced mitochondrial alterations both in LLC-PK1 cells (loss of membrane potential) and in isolated mitochondria (inhibition of mitochondrial calcium uptake, release of cytochrome c, and decrease in GSH content, aconitase activity, adenosine triphosphate (ATP) content and oxygen consumption). It is concluded that the protection exerted by SFN on mitochondrial alterations and NQO1 and γGCL enzymes may be involved in the renoprotection of SFN against CIS.
Assuntos
Anticarcinógenos/farmacologia , Antineoplásicos/toxicidade , Cisplatino/toxicidade , Glutamato-Cisteína Ligase/metabolismo , Rim/efeitos dos fármacos , Células LLC-PK1/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Doenças Mitocondriais/prevenção & controle , NAD(P)H Desidrogenase (Quinona)/metabolismo , Tiocianatos/farmacologia , Trifosfato de Adenosina/metabolismo , Animais , Sinalização do Cálcio/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Citocromos c/metabolismo , Inibidores Enzimáticos/farmacologia , Glutamato-Cisteína Ligase/antagonistas & inibidores , Glutationa/metabolismo , Isotiocianatos , Rim/metabolismo , Rim/patologia , Células LLC-PK1/metabolismo , Células LLC-PK1/patologia , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/enzimologia , NAD(P)H Desidrogenase (Quinona)/antagonistas & inibidores , Consumo de Oxigênio , Ratos , Sulfóxidos , SuínosRESUMO
Aminoglycosides are widely used in the treatment of gram-negative bacterial infections. Gentamicin (GE) acts mainly in proximal tubular cells, where it is uptake via organic anion transport system and it induces a high incidence of nephrotoxicity, which is characterized by tubular necrosis [5] leading to acute renal failure in 10 to 50% of patients. Gram-negative bacteria has lipopolysaccharide (LPS) which is an endotoxin that cause renal damage. [1] Moreover, many patients are undergone exams using radiologic contrast, which is a risk factor to induce a hemodynamic change in the kidney and to develop acute renal failure. [6] Intracellular calcium [Ca2+]i is involved in renal cellular injury [7,3] and maybe mediate the effects provoked by these drugs. This study was performed to evaluate necrosis, apoptosis, and intracellular calcium levels ([Ca2+]i) in LLC-PK1 (epithelial cell line from pig kidney) induced by GE associated with LPS and a low-osmolality media, Hexabrix (HE).
Assuntos
Antibacterianos/efeitos adversos , Antibacterianos/farmacologia , Meios de Contraste/efeitos adversos , Meios de Contraste/farmacologia , Gentamicinas/efeitos adversos , Gentamicinas/farmacologia , Ácido Ioxáglico/efeitos adversos , Ácido Ioxáglico/farmacologia , Nefropatias/induzido quimicamente , Túbulos Renais Proximais/efeitos dos fármacos , Células LLC-PK1/efeitos dos fármacos , Lipopolissacarídeos/efeitos adversos , Lipopolissacarídeos/farmacologia , Animais , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Cálcio/análise , Modelos Animais de Doenças , Técnicas In Vitro , Líquido Intracelular/química , Líquido Intracelular/efeitos dos fármacos , Nefropatias/patologia , Nefropatias/fisiopatologia , Túbulos Renais Proximais/patologia , Túbulos Renais Proximais/fisiopatologia , Células LLC-PK1/patologia , Células LLC-PK1/fisiologia , Necrose , Suínos , Fatores de TempoRESUMO
Aminoglycosides are widely used in the treatment of gram-negative bacterial infections. Gentamicin (GE) acts mainly in proximal tubular cells, where it is uptake via organic anion transport system and it induces a high incidence of nephrotoxicity, which is characterized by tubular necrosis leading to acute renal failure in 10 to 50% of patients. Gram-negative bacteria have lipopolysaccharide (LPS) which is an endotoxin that causes renal damage. Moreover, many patients are undergone exams using radiologic contrast, which is a risk factor to induce a hemodynamic change in the kidney and to develop acute renal failure. Intracellular calcium [Ca2+]i is involved in renal cellular injury and maybe mediate the effects provoked by these drugs. This study was performed to evaluate necrosis, apoptosis and intracellular calcium levels ([Ca2+]i) in LLC-PK1 (epithelial cell line from pig kidney) induced by GE associated with LPS and a low-osmolality media, Hexabrix (HE).