Gene expression and synthesis of natriuretic peptides by cultured human glomerular cells.
BACKGROUND
Atrial natriuretic peptide, brain natriuretic peptide and C-type natriuretic peptide belong to a family of hormones that have natriuretic and vasodepressor activity and may play a pathophysiologic role in hypertension, heart failure and renal failure. Whereas immunoreactive human forms of these three natriuretic peptides are found in renal tubules, it is not clear whether they are derived from the systemic circulation or from local production.


OBJECTIVE
To examine the gene expression of natriuretic peptides in cultured human glomerular cells.


METHODS
We sought to determine the presence of messenger RNA encoding for these natriuretic peptides using polymerase chain reaction following reverse transcription. The polymerase chain reaction products were confirmed by direct sequencing. Atrial natriuretic peptide, brain natriuretic peptide and C-type natriuretic peptide in cell-culture supernatants were measured by radioimmunoassays (with detection limits of 2.1, 2.1 and 0.21 pmol/l, respectively).


RESULTS
Atrial natriuretic peptide messenger RNA was not found in mesangial or glomerular epithelial cells (despite stimulation with tumor necrosis factor-alpha) except when the cells were cultured with a high concentration of fetal bovine serum (> 10%). Similarly, this peptide was not detected in supernatant unless the cells were cultured with fetal bovine serum at concentrations of > 10%. Brain natriuretic peptide messenger RNA was readily detected in cultured mesangial and glomerular epithelial cells with a lower concentration in the former. Brain natriuretic peptide was not found in the supernatant of resting mesangial cells but became detectable when incubated with tumor necrosis factor-alpha or fetal bovine serum. C-type natriuretic peptide messenger RNA was detected in mesangial and glomerular epithelial cells with a higher concentration in the latter. C-type natriuretic peptide was detected in the supernatant of resting glomerular epithelial cells and levels rose when incubated with increasing concentrations of tumor necrosis factor-alpha or fetal bovine serum. However, C-type natriuretic peptide was not detected in the supernatant of resting mesangial cells and remained undetectable following incubation with tumor necrosis factor-alpha or fetal bovine serum.


CONCLUSIONS
Our results suggest differences in the synthesis of natriuretic peptides between glomerular mesangial and epithelial cells.
Department of Medicine, University of Hong Kong, Queen Mary Hospital, Hong Kong. knlai@hkucc.hku.hk
K N
KN
Lai
J C
JC
Leung
T G
TG
Yandle
S
S
Fisher
M G
MG
Nicholls
D009320:Atrial Natriuretic Factor / Q000096:biosynthesis* / Q000235:genetics; D002478:Cells, Cultured; D003609:Dactinomycin / Q000494:pharmacology; D004847:Epithelial Cells / Q000378:metabolism / Q000502:physiology; D015870:Gene Expression; D005920:Glomerular Mesangium / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D020097:Natriuretic Peptide, Brain / Q000096:biosynthesis* / Q000235:genetics; D020098:Natriuretic Peptide, C-Type / Q000096:biosynthesis* / Q000235:genetics; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011691:Puromycin / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism
D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D020097:Natriuretic Peptide, Brain; D020098:Natriuretic Peptide, C-Type; D003609:Dactinomycin; D011691:Puromycin; D009320:Atrial Natriuretic Factor