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Malarial pigment enhances heat shock protein-27 in THP-1 cells:new perspectives for in vitro studies on monocyte apoptosis prevention

机译:疟疾色素增强THP-1细胞中的热休克蛋白27:预防单核细胞凋亡的体外研究的新观点

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摘要

Objective: To investigate the effect of malarial pigment (hemozoin, HZ) on expression of heat shock proteins (HSPs) and cell viability in human monocytes by using a stable cell line (THP-1 cells). Methods: THP-1 cells were fed with native HZ or treated with pro-apoptotic molecule gliotoxin for 9 h. Thereafter, the protein expression of HSP-27 and HSP-70 was evaluated by western blotting. Alternatively, HZ-fed cells were cultured up to 72 h and cell viability parameters (survival, apoptosis and necrosis rates) were measured by flow cytometric analysis.Results:HZ increased basal protein levels of HSP-27 without altering those of HSP-70 in THP-1 cells, and promoted long-term cell survival without inducing apoptosis. As expected, gliotoxin inhibited HSP-27 protein expression and promoted long-term cell apoptosis. Conclusions:Present data show that HZ prevents cell apoptosis and enhances the expression of anti-apoptotic HSP-27 in THP-1 cells, confirming the previous evidences obtained from HZ-fed immunopurified monocytes. Since the use of a stable cell line is pivotal to perform HSP-27 silencing experiments, monocytic THP-1 cells could be a good candidate line for such an approach, which is heavily required to clarify the role of HSP-27 in survival of impaired HZ-fed monocytes during falciparum malaria.
机译:目的:使用稳定的细胞系(THP-1细胞),研究疟疾色素(hemozoin,HZ)对人单核细胞热休克蛋白(HSPs)表达和细胞活力的影响。方法:THP-1细胞用天然HZ喂养或用促凋亡分子胶体毒素处理9小时。之后,通过蛋白质印迹法评估HSP-27和HSP-70的蛋白表达。或者,将HZ喂养的细胞培养72小时,并通过流式细胞仪分析测量细胞活力参数(存活,凋亡和坏死率)。结果:HZ增加了HSP-27的基础蛋白水平,而没有改变HSP-70的基础蛋白水平。 THP-1细胞,并在不诱导凋亡的情况下促进了长期细胞存活。如预期的那样,胶质毒素抑制HSP-27蛋白表达并促进长期细胞凋亡。结论:目前的数据表明,HZ可以防止细胞凋亡并增强THP-1细胞中抗凋亡的HSP-27的表达,证实了从HZ喂养的免疫纯化单核细胞获得的先前证据。由于使用稳定的细胞系对于执行HSP-27沉默实验至关重要,因此单核细胞THP-1细胞可能是这种方法的良好候选系,这对于阐明HSP-27在受损患者的生存中的作用非常重要。恶性疟疾期间HZ喂养的单核细胞。

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    《亚太热带医药杂志(英文版)》 |2010年第012期|934-938|共5页
  • 作者单位

    Department of Genetics, Biology and Biochemistry, University of Torino Medical School, Torino, Italy;

    Department of Genetics, Biology and Biochemistry, University of Torino Medical School, Torino, Italy;

    Department of Genetics, Biology and Biochemistry, University of Torino Medical School, Torino, Italy;

    Department of Genetics, Biology and Biochemistry, University of Torino Medical School, Torino, Italy;

    Department of Genetics, Biology and Biochemistry, University of Torino Medical School, Torino, Italy;

    Department of Genetics, Biology and Biochemistry, University of Torino Medical School, Torino, Italy;

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