Isolated glomeruli and cultured mesangial cells as in vitro models to study immunosuppressive agents

M. Potier, B. L'Azou, J. Cambar
Cell Biol Toxicol. 1996-12-01; 12(4-6): 263-270
DOI: 10.1007/bf00438156

PubMed

1. Cell Biol Toxicol. 1996 Dec;12(4-6):263-70. doi: 10.1007/BF00438156.

Isolated glomeruli and cultured mesangial cells as in vitro models to study
immunosuppressive agents.

Potier M(1), L’Azou B, Cambar J.

Author information:
(1)Laboratoire de Biologie Cellulaire, UFR des Sciences Pharmaceutiques,
Bordeaux, France.

Immunosuppressive agents, such as cyclosporin A (CsA), by their vasoconstrictive
properties, induce in vivo in patients and rodents a dramatic fall in renal
hemodynamics. The aim of this study is to review the ability of some
physiological and/or pharmacological agents which are supposed to be involved in
the renal physiopathology of CsA to prevent the contraction induced by CsA in
two in vitro glomerular models. Isolated glomeruli are obtained by a sieving
method from male Sprague-Dawley rat superficial cortex. Mesangial cells from
these isolated glomeruli are cultured in RPM1 1640 medium with 20% FCS in 5% CO2
atmosphere. The area of isolated glomeruli and cultured mesangial cells is
assessed by an image analyzer with a video camera. Each glomerulus and cell is
its own control and is photographed before incubation with any drug (T0) and
then during incubation at 5, 10, 20, and 30 min. Incubations are performed
during 30 min with 10(-6) mol/L CsA either with a 10 min pretreatment with the
vasoactive agent or without pretreatment. CsA alone induces a time- and
dose-dependent decrease in glomerular structure area (-4.7% at 10 min, -10.3% at
20 min, and -12.0% at 30 min for isolated glomeruli); Cremophore excipient or
control solute does not induce any significant decrease in surface area. CsA
with 10(-6) mol/L verapamil pretreatment induces only a slight decrease: -1.5%
at 10 min, -3.0% at 20 min, and -4.8% at 30 min. Calcium blockers nifedipine and
felodipine produce similar results. Likewise, with 10(-8) mol/L prostacyclin
analog (iloprost), only a slight area decrease in mesangial cells is noted:
-1.3% at 5 min, -1.8% at 10 min, and -3.3% at 20 min; with 10(-6) mol/L TXA2
synthesis inhibitor (CGS 12970) the results are -2.0% at 10 min, -3.6% at 20
min, and -4.3% at 30 min. Finally, a similar protective effect can be noted with
10(-5) mol/L theophylline: -0.4; -1.5 and -1.9% at 10, 20, and 30 min. In
conclusion, CsA-induced contraction in two in vitro glomerular models can be
partially or even totally prevented by pretreatment with various pharmacological
agents.