Cyclosporin increases the density of angiotensin II subtype 1 (AT1) receptors in mouse medullary thick ascending limb cells.
BACKGROUND
Cyclosporin A (CsA), a potent immunosuppressive agent, can be nephrotoxic. Because clinical studies have suggested that the intrarenal renin-angiotensin system may be involved in the mechanism responsible for CsA nephrotoxicity, we have analysed the effects of CsA on angiotensin II (Ang II) receptors in medullary thick ascending limb (mTAL) cells known to be sensitive to the action of CsA.


METHODS
Experiments were carried out on subcultured mouse mTAL cells. The expression of mRNA of Ang II subtype 1 and 2 (AT(1) and AT(2)) receptors was investigated using reverse transcription-polymerase chain reaction (RT-PCR). [(3)H]Ang II was used for radioligand and binding studies. Fluorimetric recordings using the fluorescent dye fura-2/AM were performed to determine the effect of CsA on the intracellular calcium ([Ca(2+)]i) content of untreated and Ang II-treated mTAL cells.


RESULTS
Subcultured mTAL cells expressed AT(1) and AT(2) Ang II receptor mRNAs, and binding studies revealed that the AT(1) receptors were the predominant Ang II receptor subtype ( approximately 90%) in mTAL cells. CsA (100 ng/ml, 24 h) increased (1.7-fold) the number of Ang II receptors (untreated, 315.8; +CsA, 543.6 fmol/mg protein) without altering the K(D) (untreated, 7.16; +CsA, 7.06 nM). CsA also significantly increased the level of [Ca(2+)]i measured in cultured mTAL cells both in the basal state (-CSA, 72.2; +CsA, 93.4 nM/10(6) cells) and in the presence of Ang II (-CSA, 97.8; +CsA, 206.3 nM/10(6) cells).


CONCLUSIONS
These findings suggest that the increase in Ang II AT(1) receptors and [Ca(2+)]i caused by CsA may be involved in the mechanism(s) responsible for CsA nephrotoxicity.
Wu
Mai-Szu
MS
Division of Nephrology, Chang Gung Memorial Hospital, Taipei, Taiwan. maisuwu@ms9.hinet.net
Yang
Chih-Wei
CW
Chang
Chiz-Tzuang
CT
Bens
Marcelle
M
Vandewalle
Alain
A
D000804:Angiotensin II / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000032:analysis; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D016572:Cyclosporine / Q000494:pharmacology* / Q000633:toxicity; D007166:Immunosuppressive Agents / Q000494:pharmacology* / Q000633:toxicity; D007679:Kidney Medulla / Q000166:cytology*; D008297:Male; D051379:Mice; D044140:Receptor, Angiotensin, Type 1 / Q000187:drug effects*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D007166:Immunosuppressive Agents; D044140:Receptor, Angiotensin, Type 1; D000804:Angiotensin II; D016572:Cyclosporine; D002118:Calcium