Granular Formation during Apoptosis in Blastocystis sp. Exposed to Metronidazole MTZReportar como inadecuado

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The role and function of the granular life cycle stage in Blastocystis sp, remains uncertain despite suggestions being made that the granules are metabolic, reproductive and lipid in nature. This present study aims to understand granular formation by triggering apoptosis in Blastocystis sp. by treating them with metronidazole MTZ. Blastocystis sp.cultures of 4 sub-types namely 1, 2, 3 and 5 when treated with 0.01 and 0.0001 mg-ml of metronidazole MTZ respectively showed many of the parasites to be both viable and apoptotic VA. Treated subtype 3 isolates exhibited the highest number of granular forms i.e. 88% p<0.001 0.0001 mg-ml and 69% p<0.01 0.01 mg-ml respectively at the 72 h in in vitro culture compared to other subtypes. These VA forms showed distinct granules using acridine orange AO and 4’,6-diamino-2-phenylindole DAPI staining with a mean per cell ranging from 5 in ST 5 to as high as 16 in ST 3. These forms showed intact mitochondria in both viable apoptotic VA and viable non-apoptotic VNA cells with a pattern of accumulation of lipid droplets corresponding to viable cells. Granular VA forms looked ultra-structurally different with prominent presence of mitochondria-like organelle MLO and a changed mitochondrial trans-membrane potential with thicker membrane and a highly convoluted inner membrane than the less dense non-viable apoptotic NVA cells. This suggests that granular formation during apoptosis is a self-regulatory mechanism to produce higher number of viable cells in response to treatment. This study directs the need to search novel chemotherapeutic approaches by incorporating these findings when developing drugs against the emerging Blastocystis sp. infections.

Autor: Devi Balkrishnan Dhurga, Kumar Suresh , Tian Chye Tan



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