■法尼醇,源自甾醇生物合成途径中的法尼基焦磷酸,是具有三个不饱和基团和四个可能的异构体的分子。白色念珠菌主要分泌反式,反式法尼醇(t,t-FOH)异构体,以其在调节各种真菌物种的毒力和调节形态转变过程中的作用而闻名。值得注意的是,真菌之间固醇生物合成的进化差异,包括白色念珠菌,锥虫通过麦角烷骨架合成甾醇,不同于胆固醇。本研究旨在评估外源添加反式的影响,反式法尼醇对亚马逊利什曼原虫的增殖能力,并鉴定其在寄生虫脂质分泌组中的存在。
■这项研究涉及添加外源反式,反式法尼醇评估其对亚马逊乳杆菌增殖的干扰。扩散,细胞周期,DNA片段化,线粒体功能被评估为反式效应的指标,反式法尼醇.此外,进行了脂质分泌组分析,专注于反式的检测,反式法尼醇和相关产品衍生自前体,焦磷酸法尼酯.采用计算机模拟分析来鉴定在利什曼原虫基因组中负责产生这些类异戊二烯的法尼烯合酶基因的序列。
■外源添加反式,发现反式法尼醇会干扰亚马逊乳杆菌的增殖,抑制细胞周期而不引起DNA片段化或线粒体功能丧失。尽管没有反式,培养上清液中的反式法尼醇,来自焦磷酸法尼酯的其他产品,特别是α-法尼烯和β-法尼烯,从培养的第四天开始检测到,继续增加,直到第十天。此外,通过计算机分析鉴定利什曼原虫基因组中的法尼烯合酶基因提供了对类异戊二烯生产的酶学基础的见解。
■这些发现共同提供了有关法尼醇对亚马逊乳杆菌的作用机制的初步见解。而反式,在脂质分泌组中未检测到反式法尼醇,α-法尼烯和β-法尼烯的存在提示寄生虫类异戊二烯代谢中的替代途径或修饰。在不诱导DNA片段化或线粒体功能障碍的情况下对增殖和细胞周期的抑制作用引发了关于外源反式影响的特定靶标和途径的问题。反式法尼醇.法尼烯合酶基因的鉴定为了解利什曼原虫中相关类异戊二烯的合成提供了分子基础。对这些机制的进一步探索可能有助于开发针对利什曼原虫感染的新型治疗策略。
Farnesol, derived from farnesyl pyrophosphate in the sterols biosynthetic pathway, is a molecule with three unsaturations and four possible isomers. Candida albicans predominantly secretes the trans, trans-farnesol (t, t-FOH) isomer, known for its role in regulating the virulence of various fungi species and modulating morphological transition processes. Notably, the evolutionary divergence in sterol biosynthesis between fungi, including Candida albicans, and trypanosomatids resulted in the synthesis of sterols with the ergostane skeleton, distinct from cholesterol. This study aims to assess the impact of exogenously added trans, trans-farnesol on the proliferative ability of Leishmania amazonensis and to identify its presence in the lipid secretome of the parasite.
The study involved the addition of exogenous trans, trans-farnesol to evaluate its interference with the proliferation of L. amazonensis promastigotes. Proliferation, cell cycle, DNA fragmentation, and mitochondrial functionality were assessed as indicators of the effects of trans, trans-farnesol. Additionally, lipid secretome analysis was conducted, focusing on the detection of trans, trans-farnesol and related products derived from the precursor, farnesyl pyrophosphate. In silico analysis was employed to identify the sequence for the farnesene synthase gene responsible for producing these isoprenoids in the Leishmania genome.
Exogenously added trans, trans-farnesol was found to interfere with the proliferation of L. amazonensis promastigotes, inhibiting the cell cycle without causing DNA fragmentation or loss of mitochondrial functionality. Despite the absence of trans, trans-farnesol in the culture supernatant, other products derived from farnesyl pyrophosphate, specifically α-farnesene and β-farnesene, were detected starting on the fourth day of culture, continuing to increase until the tenth day. Furthermore, the identification of the farnesene synthase gene in the Leishmania genome through in silico analysis provided insights into the enzymatic basis of
isoprenoid production.
The findings collectively offer the first insights into the mechanism of action of farnesol on L. amazonensis. While trans, trans-farnesol was not detected in the lipid secretome, the presence of α-farnesene and β-farnesene suggests alternative pathways or modifications in the
isoprenoid metabolism of the parasite. The inhibitory effects on proliferation and cell cycle without inducing DNA fragmentation or mitochondrial dysfunction raise questions about the specific targets and pathways affected by exogenous trans, trans-farnesol. The identification of the farnesene synthase gene provides a molecular basis for understanding the synthesis of related isoprenoids in Leishmania. Further exploration of these mechanisms may contribute to the development of novel therapeutic strategies against Leishmania infections.