[Effect of aldosterone on the expression of plasminogen activator inhibitor-1 in renal mesangial cells: experiment with rat renal mesangial cells].
OBJECTIVE
To evaluate the effects of aldosterone (ald) on the expression of plasminogen activator inhibitor-1 (PAI-1) in renal mesangial cells (MCs) and the changes after aldosterone was blocked with spironolactone.


METHODS
Rat MCs of the line HBZY-1 were cultured and divided into 5 groups: ald group, treated with aldosterone (1 micromol/L or 100 nmol/L), ald combined with spironolactone 1 nmol/L for 24 hours, and control group. Anotfher cells were cultured and treated with ald 100 nmol/L for 0, 0.5, 1, 2, 4, 6, 8, and 24 h respectively or treated with ald of the concentrations of 10(-5), 10(-6), 10(-7), 10(-8), 10(-9), 10(-10), pr 10(-11) mol/L. RT-PCR and Western blotting were used to detect the mRNA and protein expression of PAI-1. Confocal laser scanning microscopy was used to detect the ROS level in the MCs. The transforming growth factor-beta1 (TGF-beta1) level in the medium was detected by ELISA.


RESULTS
The PAI-1 mRNA and protein levels of the ald 100 nmol/L and 1 micromol/L groups became 2 times and 1.8 times, and 1.7 times and 1.9 times respectively those of the control group (all P < 0.01). The PAI-1 mRNA and protein levels of the ald + spirolactone group were not significantly different from those of the control group (all P > 0.05). The TGF-beta1 levels of the 100 nmol/L and 1 micromol/L aldosterone groups were 147 pg/ml +/- 27 pg/ml and 183 pg/ml +/- 25 pg/ml respectively, both significantly higher than that of the control group (75 pg/ml +/- 23 pg/ml, both P < 0.01). The TGF-beta1 levels of the groups with spirolactone were not significantly different from that of the control group (both P > 0.05). The ROS levels of the 1 micromol/L and 100 nmol/L ald groups were 4.87 times and 4.77 times that of the control group (both P < 0.01), and the ROS levels of the groups with spirolactone were 1.95 time and 1.66 times that of the control group (both P > 0.05). 100 nmol/L ald time-dependently increased the PAI-1 level since 4 hours after exposure to ald (all P < 0.05). Aldosterone dose-dependently increased the PAI-1 level (all P < 0.05).


CONCLUSIONS
Aldosterone upregulates the production of PAI-1 by MCs and increases the expression of TGF-beta1 and cellular ROS in the MCs significantly, which is mediated through mineralocorticoid receptor.
Department of Nephrology, Renmin Hospital, Wuhan University, Wuhan 430060, China.
Jun
J
Yuan
Ru-han
RH
Jia
Yan
Y
Bao
D000450:Aldosterone / Q000494:pharmacology*; D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression / Q000187:drug effects; D050527:Mesangial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018613:Microscopy, Confocal; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013148:Spironolactone / Q000494:pharmacology; D013997:Time Factors; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
D017395:Plasminogen Activator Inhibitor 1; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D053773:Transforming Growth Factor beta1; D013148:Spironolactone; D000450:Aldosterone