[Antioxidant effects of mast cell inhibitors in a human conjunctival cell line].
OBJECTIVE
The aim of this study was to evaluate in vitro antioxidant effects of two mast cells inhibitors.


METHODS
Cytotoxicity tests were done on a continuous human conjunctival cell line using microplate cold light cytofluorimetry. Membrane integrity (neutral red test), DNA condensation (Hoechst 33342 test), and reactive oxygen species (ROS) production (dichlorofluoresceine diacetate and hydroethidine tests) were evaluated on living cells treated with sodium cromoglycate and N-acetyl-aspartyl glutamic acid (NAAGA) preserved (benzalkonium chloride: BAC at 0.01%) and unpreserved after 60 minutes of treatment or 60 minutes and 24 hours of cell recovery. They were tested pure and at a 1/10 dilution. ROS production was also evaluated after a 60 minute pretreatment with antiallergic drugs and a 15-minute treatment with BAC, according to previous experiments performed on BAC showing its ROS production properties.


RESULTS
No cytotoxicity was observed with the unpreserved formulations of antiallergic drugs. An apoptotic phenomenom was suggested with preserved drugs after a 1-hour treatment, whereas a necrotic mechanism appeared after a 24-hour cell recovery period. A ROS production decrease was observed with the two preserved and unpreserved drugs tested (p<0.001 compared to BAC) even if it was significantly higher with cromoglycate formulations. A ROS production decrease also was detected after a pretreatment with antiallergic drugs and treatment with BAC (p<0.001 compared to BAC alone).


CONCLUSIONS
In vitro, no cytotoxicity was found with the two unpreserved mast cell inhibitors tested. An antioxidant effect also was observed with these two molecules; sodium cromoglycate appeared to be the best free radical scavenger.
Debbasch
C
C
Unité de Pharmaco-Toxicologie Cellulaire, CHNO des Quinze-Vingts, 28, rue de Charenton, 75012 Paris.
Pisella
P J
PJ
Rat
P
P
Warnet
J M
JM
Baudouin
C
C
D018926:Anti-Allergic Agents / Q000494:pharmacology*; D018927:Anti-Asthmatic Agents / Q000494:pharmacology*; D000975:Antioxidants; D017209:Apoptosis; D001548:Benzalkonium Compounds / Q000494:pharmacology; D002460:Cell Line; D003228:Conjunctiva / Q000166:cytology / Q000187:drug effects*; D004205:Cromolyn Sodium / Q000494:pharmacology*; D003601:Cytotoxicity Tests, Immunologic; D004151:Dipeptides / Q000494:pharmacology; D004847:Epithelial Cells / Q000187:drug effects; D005434:Flow Cytometry; D016166:Free Radical Scavengers; D006801:Humans; D008407:Mast Cells / Q000187:drug effects*; D009336:Necrosis; D017382:Reactive Oxygen Species; D013997:Time Factors
D018926:Anti-Allergic Agents; D018927:Anti-Asthmatic Agents; D000975:Antioxidants; D001548:Benzalkonium Compounds; D004151:Dipeptides; D016166:Free Radical Scavengers; D017382:Reactive Oxygen Species; C027172:isospaglumic acid; D004205:Cromolyn Sodium