Cellular Changes Induced by Kinin B-1 Receptor Deletion: Study of Endothelial Nitric Oxide Metabolism

Autor(es)

Loiola, Rodrigo A. ^[1] ; Torres, Tathiany C. ^[1] ; Landgraf, Richardt G. ^[1] ; Pesquero, Joao Bosco ^[2] ; Fernandes, Liliam ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed Sao Paulo, Lab Inflamacao & Farmacol Vasc, Inst Ciencias Ambientais Quim & Farmaceut, BR-09913030 Diadema, SP - Brazil

^[2] Univ Fed Sao Paulo, Dept Biofis, BR-04039032 Sao Paulo, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: INTERNATIONAL JOURNAL OF PEPTIDE RESEARCH AND THERAPEUTICS; v. 21, n. 3, p. 375-382, SEP 2015.
• Citações Web of Science: 1

Resumo

Kinins are vasoactive peptides involved in endothelial function and vascular tonus. The present study determined the influence of the kinin B-1 receptor subtype on endothelial nitric oxide (NO) metabolism by using primary cultured cells obtained from B-1 knockout (B (1) (-/-) ) and Wild Type (WT) mice. By using specific fluorescent dyes, NO and superoxide anion ( (a (TM))) production was determined in absence or presence of ascorbate or tetrahydrobiopterin (BH4). The activity of the enzyme superoxide dismutase (SOD) was determined by immune enzyme assay, and endothelial NOS (eNOS) activation was analyzed through expression of phospho-eNOS (p-eNOS, Ser1177) by western blot. The NO release {[}quantified by densitometry and expressed as arbitrary units (a.u.)] was markedly reduced in B (1) (-/-) (35.8 +/- A 3.1{*} a.u.) when compared to WT cells (66.9 +/- A 3.2 a.u.); this impaired response was reversed by ascorbate (101.8 +/- A 6.0 a.u.) and BH4 (54.3 +/- A 1.7 a.u.). B (1) (-/-) cells showed a marked increase in (a (TM)) production (77.1 +/- A 2.5{*} a.u.) versus WT (29.3 +/- A 6.9 a.u.), which was reversed by ascorbate (35.3 +/- A 6.4 a.u.), but not by BH4. SOD activity was similar between groups, and B (1) (-/-) cells presented a significant reduction in the expression of p-eNOS. In conclusion, the reduced NO availability detected in B (1) (-/-) cells appears to be related to a recurrent process involving BH4 oxidation, NOS uncoupling and further enhancement of NOS-derived (a (TM)). B-1 receptor deletion also impairs the phosphorylation of eNOS at the Ser1177 residue, contributing to the deficient NO production at the endothelial level. (AU)

• Processo FAPESP: 08/06676-8 - Biologia celular e molecular dos sistemas calicreínas-cininas e renina-angiotensina
• Beneficiário: João Bosco Pesquero
• Linha de fomento: Auxílio à Pesquisa - Temático
• Processo FAPESP: 07/59039-2 - Participação de receptores de cininas em vias de sinalização ativadas por angiotensina: aspectos vasculares, bioquímicos e moleculares
• Beneficiário: Liliam Fernandes
• Linha de fomento: Auxílio à Pesquisa - Apoio a Jovens Pesquisadores
• Processo FAPESP: 10/01404-0 - Estudos in vivo e in vitro da participação da leptina em diferentes modelos de inflamação pulmonar: mediadores inflamatórios e vias de sinalização envolvidas
• Beneficiário: Richardt Gama Landgraf
• Linha de fomento: Auxílio à Pesquisa - Apoio a Jovens Pesquisadores
• Processo FAPESP: 14/18760-4 - Fatores endoteliais venosos: influência de angiotensina II e endotelina-1
• Beneficiário: Liliam Fernandes
• Linha de fomento: Auxílio à Pesquisa - Regular