Activation of purinergic P2Y2 receptors inhibits inducible NO synthase in cultured rat mesangial cells
1998
Cytokine-induced nitric oxide (NO) is produced on glomerular inflammation. Glomerular injury and thrombocyte aggregation result in the release of nucleotides, which may regulate induced NO synthesis in cultured rat mesangial cells (MCs). ATP (10 -3 M) inhibited 24-h nitrite production induced by lipopolysaccharide (LPS, 10 μg/ml)/interferon-γ (IFN-γ, 100 U/ml) by 48.2 ± 6.3%, as well as induction of inducible NOS (iNOS) protein and mRNA. Also, coincubation with either 10 -4 M of UTP, ATP, or ATP γ S inhibited LPS/IFN-γ-induced nitrite production by 29.9 ± 5.8, 36.4 ± 4.3, and 50.3 ± 6.5%, respectively, indicating involvement of purinergic P2Y2 receptors. Correspondingly, cultured MCs expressed P2Y2 receptor mRNA. Agonists for other purinergic receptors [α,β-methylene-ATP, 3'-O-(4-benzoyl)-benzoyl-ATP, 2-methylthio-ATP, ADP, UDP, adenosine] were ineffective. Treatment with the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA, 10 -8 M) reproduced the inhibitory effect of ATP on iNOS protein expression and nitrite inhibition (by 46.6± ± 10.4%). The effect of ATP or PMA was reversed by the PKC inhibitors Ro-31-8220 (10 -8 M) and 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (10 -5 M), indicating that suppression of iNOS is mediated via activation of PKC through stimulated P2Y2 receptors. In conclusion, the release of purine mediators may play a critical role for iNOS expression and synthesis of NO during glomerular inflammatory disorders.
Keywords:
- Correction
- Source
- Cite
- Save
- Machine Reading By IdeaReader
49
References
30
Citations
NaN
KQI