摘要
Objective:To investigate the regulation of matrix metalloproteinases(MMPs) and tissue inhibitors of melalloproleinases(TIMPs) in human microvascular endothelium(HMEC-1) exposed to erythrocytes infected by different strains of Plasmodium falciparum(P.falciparum).Methods: HMEC—1 eells were co—incubated for 72 h with erythrocytes infected by late stage trophozoite of D10(chloroquine-sensilive) or W 2(chloroquine-resistant) P.falciparum strains.Cell supernatants were then collected and the levels of pro- or active gelatinases MMP-9 and MMP-2 were evaluated by gelatin zymograpln and densitometry.The release of pro-MMP-9,MMP-3.MMP-1 and TIMP-1 proteins was analyzed by western blotting and densitometry.Results:Infected erythrocytes induced de novo proMMP-9 and MMP-9 release.Neither basal levels of proMMP-2 were altered,nor active MMP-2 was found.MMP-3 and MMP-1 secretion was significant!) enhanced,whereas basal TIMP-1 was unaffected.All effects were similar for both strains. Conclusions:P.falciparum parasites,either chloroquine-sonsitive or -resistant,induce the release of active MMP-9 protein from human microvascular endothelium,by impairing balances between proMMP-9 and its inhibitor,and by enhancing the levels of its activators.This work provides new evidence on MMP involvement in malaria,pointing at MMP-9 as a possible target in adjuvant therapy.
Objective:To investigate the regulation of matrix metalloproteinases(MMPs) and tissue inhibitors of melalloproleinases(TIMPs) in human microvascular endothelium(HMEC-1) exposed to erythrocytes infected by different strains of Plasmodium falciparum(P.falciparum).Methods: HMEC—1 eells were co—incubated for 72 h with erythrocytes infected by late stage trophozoite of D10(chloroquine-sensilive) or W 2(chloroquine-resistant) P.falciparum strains.Cell supernatants were then collected and the levels of pro- or active gelatinases MMP-9 and MMP-2 were evaluated by gelatin zymograpln and densitometry.The release of pro-MMP-9,MMP-3.MMP-1 and TIMP-1 proteins was analyzed by western blotting and densitometry.Results:Infected erythrocytes induced de novo proMMP-9 and MMP-9 release.Neither basal levels of proMMP-2 were altered,nor active MMP-2 was found.MMP-3 and MMP-1 secretion was significant!) enhanced,whereas basal TIMP-1 was unaffected.All effects were similar for both strains. Conclusions:P.falciparum parasites,either chloroquine-sonsitive or -resistant,induce the release of active MMP-9 protein from human microvascular endothelium,by impairing balances between proMMP-9 and its inhibitor,and by enhancing the levels of its activators.This work provides new evidence on MMP involvement in malaria,pointing at MMP-9 as a possible target in adjuvant therapy.
基金
supported by Universita di Milano(PUR.2009) to Nicoletta Basilico and Charity Funds from Mrs.Franca Squazza to Mauro Prato
Mauro Prato holds a professorshipgranted by Universita Torino and Azienda Sanitaria Locale-19(ASL-19)
