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氯卟啉E6对硕大利什曼原虫和巴西利什曼原虫前鞭毛体的光毒性——体外研究

Chlorin E6 phototoxicity in L. major and L. braziliensis promastigotes-In vitro study.

作者信息

Pinto Juliana Guerra, Pereira André Henrique Correia, de Oliveira Marco Antonio, Kurachi Cristina, Raniero Leandro José, Ferreira-Strixino Juliana

机构信息

Laboratório de Terapia Fotodinâmica, Instituto de Pesquisa e desenvolvimento, Univap-Universidade do Vale do Paraíba, Av. Shishima Hifumi, 2911, 12244-000 São José dos Campos, SP, Brazil.

Laboratório de Terapia Fotodinâmica, Instituto de Pesquisa e desenvolvimento, Univap-Universidade do Vale do Paraíba, Av. Shishima Hifumi, 2911, 12244-000 São José dos Campos, SP, Brazil.

出版信息

Photodiagnosis Photodyn Ther. 2016 Sep;15:19-24. doi: 10.1016/j.pdpdt.2016.04.014. Epub 2016 May 6.

Abstract

BACKGROUND

Cutaneous leishmaniasis is a zoonosis caused by protozoa of the genus Leishmania. Conventional treatments are long and aggressive, and they trigger a diversity of side effects. Photodynamic Therapy was originally proposed as a treatment for cancer, and it now appears to be a promising therapy for local treatment with fewer side effects of infectious diseases.

METHODS

This study aimed to evaluate Chlorin e6 internalization by Leishmania major and Leishmania braziliensis promastigotes and its viability and effects on mitochondrial activity. Control groups were kept in the dark, while PDT groups received fluence of 10J/cm(2) (660nm). Chlorin internalization was evaluated using confocal microscopy after one hour of incubation for both species.

RESULTS

The mitochondrial activity was evaluated by MTT assay, and viability was measured by the Trypan blue exclusion test. Giemsa staining was used to observe morphological changes. PS was internalized in both species and mitochondrial activity changed in all groups. However, the obtained MTT and Trypan results indicated that despite the change in mitochondrial activity in the dark groups, their viability was not affected, whereas the PDT treated groups had significantly reduced viability. Morphology was drastically altered in PDT treated groups, while groups kept in the dark exhibited the standard morphology.

CONCLUSIONS

This study demonstrates that Chlorin has great potential for being used in PDT as a treatment for cutaneous leishmaniasis, although more studies are needed to determine in vivo application protocols.

摘要

背景

皮肤利什曼病是由利什曼原虫属的原生动物引起的一种人畜共患病。传统治疗方法疗程长且具有侵袭性,还会引发多种副作用。光动力疗法最初被提议用于癌症治疗,现在看来它是一种有前景的局部治疗方法,对传染病的副作用较少。

方法

本研究旨在评估氯e6被硕大利什曼原虫和巴西利什曼原虫前鞭毛体摄取的情况及其对线粒体活性的活力和影响。对照组置于黑暗中,而光动力疗法组接受10J/cm²(660nm)的能量密度照射。两种原虫在孵育一小时后,使用共聚焦显微镜评估氯的摄取情况。

结果

通过MTT法评估线粒体活性,用台盼蓝排斥试验测量活力。使用吉姆萨染色观察形态变化。两种原虫均摄取了光敏剂,所有组的线粒体活性均发生了变化。然而,获得的MTT和台盼蓝结果表明,尽管黑暗组的线粒体活性发生了变化,但其活力并未受到影响,而光动力疗法治疗组的活力显著降低。光动力疗法治疗组的形态发生了剧烈改变,而置于黑暗中的组呈现出标准形态。

结论

本研究表明,氯在用于光动力疗法治疗皮肤利什曼病方面具有巨大潜力,尽管还需要更多研究来确定体内应用方案。

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