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LPS and hypoxia alters the production/release of catecholamines in Immortalized Human Mesangial Cell
Author(s) -
Zanini Ana Paula,
Arita Danielle Yuri,
Almeida Waldemar Silva,
Casarini Dulce Elena
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.1003.4
Subject(s) - hypoxia (environmental) , intracellular , lipopolysaccharide , endocrinology , medicine , chemistry , mesangial cell , catecholamine , blot , tyrosine hydroxylase , dopamine , biology , kidney , biochemistry , organic chemistry , oxygen , gene
Catecholamines (CA)‐dopamine (DA), L‐Dopa, epinephrine (EP) and norepinephrine‐are involved in filtration and renal vasoconstriction processes, and can be produced by mesangial cells (MC). MC expresses Toll Like Receptor 4, that detects lipopolysaccharide (LPS). LPS and hypoxia, both present in sepsis, can lead to acute renal failure. The aim was to asses the effect of LPS and hypoxia on the production/release of CA in immortalized human mesangial cell (IHMC). IHMC were cultured in DMEM and incubated with LPS (0.1, 1.0, 5.0 and 100 μg/mL) for 2 hours, or exposed to hypoxia for 30 min. CA were extracted from the medium and cell lysates was quantified by HPLC‐ED. Tyrosine hydroxylase (TH) expression ‐ step‐limiting enzyme ‐ was evaluated by Western blotting. LPS at 1 μg/mL induced a decline in intracellular L‐Dopa level as compared with control (5.3±1.6 vs. 10.8±1.0 pg/mg protein). The concentration of 5 μg/mL decreased the intracellular level of L‐Dopa (4.9±1.3 vs. 10.3±1.0 pg/mg protein) and increased EP concentration (8.8±1.9 vs. 2.2±0.5 pg/mg protein). LPS at 100 μg/mL increased the levels of intracellular EP (102.7±8.4 vs. 65.8±10.1 pg/mg), L‐Dopa (48.5±2.4 vs. 22.2±4.1 pg/mg protein) and DA (24.9±2.9 vs. 15.9±1.4 pg/mg protein) and decreased the concentration of EP (125.2±13.7 vs. 205.4±10.4 pg/mg protein) and L‐Dopa (34.6±5.7 vs. 57.4±8.6 pg/mg protein) in extracellular. TH expression was significantly affected by LPS (100 μg/ml), however it did not occurr under hypoxia condition. These results suggest that LPS can influence the production/release of CA in IHMC. Support by CNPq.