B. L'Azou et al., In vitro models to study mechanisms involved in cyclosporine A-mediated glomerular contraction (vol 73, pg 337, 1999), ARCH TOXIC, 73(8-9), 1999, pp. NIL_1
The immunosuppressive drug, cyclosporine A (CsA), which is successfully use
d to prevent rejection in organ transplantation, induces renal side-effects
as shown by a decrease in glomerular filtration rate and ultrafiltration c
oefficient regulated by the tone of mesangial cells.
The aim of the present study was to investigate the effect of CsA on isolat
ed glomeruli and mesangial cells, which constitute appropriate in vitro mod
els for renal vasoreactivity studies. The roles of different intracellular
and extracellular mediators such as calcium, endothelin-1 (ET-I), prostagla
ndins (TXA(2) and PGI(2)) and reactive oxygen intermediates (ROIs) were ana
lysed.
CsA caused a concentration- and time-dependent decrease in the planar cross
-sectional areas of isolated glomeruli and mesangial cells as determined by
image analysis. Intracytosolic free calcium concentration determined by fl
uorimetric analysis was significantly increased after 30 min CsA (10 mu M)
incubation. In the contraction experiment, the calcium antagonist verapamil
inhibited the CsA response. ET-1, TXB2 and keto-PGF(1 alpha) were determin
ed directly, however no changes were found statistically significantly diff
erent from respective controls. In contrast to these results, the ET-1 spec
ific antibody was able to reduce CsA-mediated cell contraction. In the pres
ence of a prostacyclin agonist iloprost, CsA-induced contraction was also m
odified. The role of ROIs using a 2'7'-dichlorofluorescein diacetate (DCFdA
c) fluorimetric method was directly determined by observing, with 10 mu M C
sA, a significant production of hydrogen peroxide (H2O2), which was able al
one to induce mesangial cell contraction. Preincubation with the antioxidan
ts led to a significant inhibition of mesangial cell contraction. These res
ults suggest that CsA caused an imbalance in the normal level of all invest
igated vasoconstrictive and vasodilator mediators, which shifted towards th
e advantage of vasoconstrictive action.