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Expression of inducible nitric oxide synthase, caspase-3 and production of reactive oxygen intermediate on endothelial cells culture (HUVECs) treated with <em>P. falciparum</em> infected erythrocytes and tumour necrosis factor-α

By Loeki E. Fitri

Abstract

<p>Cytoadherence of P. falciparum infected erythrocytes on endothelial cells is a key factor in development of severe malaria. This process may associated with the activation of local immune that was enhanced by tumour necrosis factor-α (TNF-α). This study was conducted to see the influence of P.falciparum infected erythrocytes cytoadherence and TNF-α treatment in inducing endothelial cells activation in vitro. inducible nitric oxide synthase (iNOS) and caspase-3 expression, also reactive oxygen intermediate (ROI) production were used as parameters. An Experimental laboratory study had been done to observe endothelial cells activation (HUVECs) after treatment with TNF-α for 20 hours or <em>P. falciparum</em> infected erythrocytes for 1 hour or both of them. Normal endothelial cells culture had been used as a control. Using immunocytochemistry local immune activation of endothelial cells was determined by iNOS and caspase-3 expression. Nitro Blue Tetrazolium reduction-assay was conducted to see the ROI production semi quantitatively. inducible nitric oxide synthase expression only found on endothelial cells culture treated with <em>P. falciparum</em> infected erythrocytes or both <em>P. falciparum</em> infected erythrocytes and TNF-α. Caspase-3 expression found slightly on normal endothelial cells culture. This expression increased significantly on endothelial cells culture treated with both P.falciparum infected erythrocytes and TNF-α (p=0.000). The normal endothelial cells release low level of ROI in the presence of non-specific trigger, PMA. In the presence of <em>P. falciparum</em> infected erythrocytes or TNF-α or both of them, some cells showed medium to high levels of ROI. Cytoadherence of <em>P. falciparum</em> infected erythrocytes and TNF α treatment on endothelial cells can induce activation of local immune marked by increase inducible nitric oxide synthase and release of free radicals that cause cell damage. <em><strong>(Med J Indones 2006; 15:151-6) </strong></em></p><p><strong>Keywords:</strong> <em>P.falciparum ,HUVECs, TNF-α, iNOS, Caspase-3, ROI</em></p

Topics: Medicine (General), R5-920, Medicine, R
Publisher: Faculty of Medicine Universitas Indonesia
Year: 2006
OAI identifier: oai:doaj.org/article:0e1314a8dbff49459e5f54eb0fa84a82
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