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| IJCEM Copyright © 2008-All rights reserved. Published by e-Century Publishing Corporation, Madison, WI 53711 |
Int J Clin Exp Med 2(3):221-232;2009.
Original Article
Sex hormones and leflunomide treatment of human macrophage cultures: effects
on apoptosis
iPaola Montagna, Renata Brizzolara, Stefano Soldano, Carmen Pizzorni, Alberto Sulli, Maurizio Cutolo
Research Laboratories and Clinical Academic Unit of Rheumatology, Department of Internal Medicine, University of Genova Italy.
Received July 31, 2009; accepted August 21, 2009; available online August 31, 2009
Abstract: In order to establish the possible gender influence on the activity of leflunomide (LEF) in the rheumatoid arthritis (RA), we
evaluated the proapoptotic activity of the active LEF metabolite A77 1726 (LEF-M), in combination with sex hormones, on cultures of
human macrophages. In particular, we focussed our investigation on the triggering phase of the apoptosis. Cultures of macrophages
from activated THP-1 cells and from RA synovial tissues were treated with LEF-M alone [30M] or in presence of 17beta-estradiol (E2)
(10-9M) or testosterone (T) (10-8M) for 24 hours. FAS, FAS-L, FADD (Fas-Associated via Death Domain) and FLICE (FADD-Like
Interleukin-1 beta Converting Enzime) were evaluated by immunocytochemistry (ICC), Western blot (WB) and reverse transcriptase-
multiplex polymerase chain reaction (RT-MPCR). Regarding macrophages from THP-1 cells (M), the ICC showed that LEF-M exerted a
significant up-regulation on all investigated apoptotic proteins, when compared to untreated cells (control) (p<0.001). On the contrary,
E2 increased significantly FAS-L positivity (p<0.05) but down-regulated significantly FADD (p<0.01), while others apoptotic proteins
were not modulated when compared to control. Regarding RA synovial macrophages (SM), the ICC showed that LEF-M exerted a
significant up-regulation on FAS and FAS-L when compared to control (p<0.001). On the contrary, E2 down-regulated significantly FAS-L
(p<0.001), while FAS was not modulated respect to control. T increased significantly the apoptotic proteins in all conditions.
The results of the present study might suggest a less efficient therapeutical effect of LEF in female patients, due to the contrasting
action of estrogens on LEF- induced apoptosis. (IJCEM907006).
Key Words: Rheumatoid arthritis, leflunomide, apoptosis, macrophages, sex hormones
Full Text PDF
Address all correspondence to:
Maurizio Cutolo MD
Research Laboratories and Academic Unit of Clinical Rheumatology
University of Genova Italy
Viale Benedetto XV, 6
16132 Genova Italy
Phone +39 010 353 7994
FAX +39 010 353 8885
Mobile (+39) 335 23 36 21
E-mail: mcutolo@unige.it
Original Article
Sex hormones and leflunomide treatment of human macrophage cultures: effects
on apoptosis
iPaola Montagna, Renata Brizzolara, Stefano Soldano, Carmen Pizzorni, Alberto Sulli, Maurizio Cutolo
Research Laboratories and Clinical Academic Unit of Rheumatology, Department of Internal Medicine, University of Genova Italy.
Received July 31, 2009; accepted August 21, 2009; available online August 31, 2009
Abstract: In order to establish the possible gender influence on the activity of leflunomide (LEF) in the rheumatoid arthritis (RA), we
evaluated the proapoptotic activity of the active LEF metabolite A77 1726 (LEF-M), in combination with sex hormones, on cultures of
human macrophages. In particular, we focussed our investigation on the triggering phase of the apoptosis. Cultures of macrophages
from activated THP-1 cells and from RA synovial tissues were treated with LEF-M alone [30M] or in presence of 17beta-estradiol (E2)
(10-9M) or testosterone (T) (10-8M) for 24 hours. FAS, FAS-L, FADD (Fas-Associated via Death Domain) and FLICE (FADD-Like
Interleukin-1 beta Converting Enzime) were evaluated by immunocytochemistry (ICC), Western blot (WB) and reverse transcriptase-
multiplex polymerase chain reaction (RT-MPCR). Regarding macrophages from THP-1 cells (M), the ICC showed that LEF-M exerted a
significant up-regulation on all investigated apoptotic proteins, when compared to untreated cells (control) (p<0.001). On the contrary,
E2 increased significantly FAS-L positivity (p<0.05) but down-regulated significantly FADD (p<0.01), while others apoptotic proteins
were not modulated when compared to control. Regarding RA synovial macrophages (SM), the ICC showed that LEF-M exerted a
significant up-regulation on FAS and FAS-L when compared to control (p<0.001). On the contrary, E2 down-regulated significantly FAS-L
(p<0.001), while FAS was not modulated respect to control. T increased significantly the apoptotic proteins in all conditions.
The results of the present study might suggest a less efficient therapeutical effect of LEF in female patients, due to the contrasting
action of estrogens on LEF- induced apoptosis. (IJCEM907006).
Key Words: Rheumatoid arthritis, leflunomide, apoptosis, macrophages, sex hormones
Full Text PDF
Address all correspondence to:
Maurizio Cutolo MD
Research Laboratories and Academic Unit of Clinical Rheumatology
University of Genova Italy
Viale Benedetto XV, 6
16132 Genova Italy
Phone +39 010 353 7994
FAX +39 010 353 8885
Mobile (+39) 335 23 36 21
E-mail: mcutolo@unige.it
