Totino Paulo Renato Rivas, Daniel-Ribeiro Cláudio Tadeu, Corte-Real Suzana, de Fátima Ferreira-da-Cruz Maria
Laboratory of Malaria Research, Instituto Oswaldo Cruz, Fiocruz, Pavilhão Leônidas Deane, Manguinhos, 21045-900 Rio de Janeiro, RJ, Brazil.
Exp Parasitol. 2008 Apr;118(4):478-86. doi: 10.1016/j.exppara.2007.10.017. Epub 2007 Dec 23.
It has been reported that an apoptotic cell death process can occur with protozoans, but no consensus on Plasmodium susceptibility to apoptosis was reached till now. Thus, we evaluated if Plasmodium falciparum blood forms undergo apoptosis after in vitro pressure with chloroquine, S-nitroso-N-acetyl-penicillamine (SNAP) or staurosporine. Inhibition of parasite growth and loss of viability were observed in treated cultures by both light microscopy and flow cytometry. When DNA fragmentation was verified, only a small number of TUNEL-positive parasites was detected in treated cultures and pretreatment of parasite with a general caspase inhibitor was not able to prevent parasite death. Considering the lack of apoptotic characteristics and the observation of parasites with cytoplasmatic vacuolization by electron microscopy, we conclude that P. falciparum parasites under chloroquine, SNAP or staurosporine pressures do not die by apoptosis but by a process similar to autophagy. The autophagic pathway could be explored as an alternative target for the development of new antimalarial drugs.
据报道,原生动物可发生凋亡性细胞死亡过程,但迄今为止关于疟原虫对凋亡的易感性尚未达成共识。因此,我们评估了恶性疟原虫血液阶段在体外受到氯喹、S-亚硝基-N-乙酰青霉胺(SNAP)或星形孢菌素作用后是否会发生凋亡。通过光学显微镜和流式细胞术在处理过的培养物中观察到了寄生虫生长的抑制和活力的丧失。当证实存在DNA片段化时,在处理过的培养物中仅检测到少量TUNEL阳性寄生虫,并且用一般的半胱天冬酶抑制剂对寄生虫进行预处理并不能阻止寄生虫死亡。考虑到缺乏凋亡特征以及通过电子显微镜观察到寄生虫出现细胞质空泡化,我们得出结论,在氯喹、SNAP或星形孢菌素压力下的恶性疟原虫寄生虫不是通过凋亡死亡,而是通过类似于自噬的过程死亡。自噬途径可作为开发新型抗疟药物的替代靶点进行探索。