Laboratory of Molecular Biology, Institute of Biological Sciences, Federal University of Goiás, Goiânia, Goiás, Brazil.
Institute of Physics, Federal University of Goiás, Goiânia, Goiás, Brazil.
J Proteomics. 2022 Aug 30;266:104683. doi: 10.1016/j.jprot.2022.104683. Epub 2022 Jul 12.
Paracoccidioides spp. are the etiological agent of paracoccidioidomycosis, a disease that causes skin lesions and affect the lungs and other organs. The current management of the disease is long and has several side effects that often lead the patient to give up the treatment, sequelae and even death. The search for new forms of treatment that minimize these drawbacks is very important. Thus, natural compounds are targets of great interest. Curcumin is one of the main components of the tubers of Curcuma longa, presenting medicinal effects well described in the literature, including the antifungal effect on Paracocidioides brasiliensis. Nevertheless, the mechanisms related to the antifungal effect of such compound are still unknown, so the objective of the present research is to understand what changes occur in the metabolism of P. brasiliensis after exposure to curcumin and to identify the main targets of the compound. Proteomic analysis as based on nanoUPLC-MS analysis and the functional classification of the identified proteins. The main metabolic processes that were being regulated were biologically validated through assays such as fluorescence microscopy, EPR and phagocytosis. Proteomic analysis revealed that curcumin regulates several metabolic processes of the fungus, including important pathways for energy production, such as the glycolytic pathway, beta oxidation and the glyoxylate cycle. Protein synthesis was down-regulated in fungi exposed to curcumin. The electron transport chain and the tricarboxylic acid cycle were also down-regulated, indicating that both the mitochondrial membrane and the mitochondrial activity were compromised. Plasma membrane and cell wall structure were altered following exposure to the compound. The fungus' ability to survive the phagocytosis process by alveolar macrophages was reduced. Thus, curcumin interferes with several metabolic pathways in the fungus that causes paracoccidioidomycosis. BIOLOGICAL SIGNIFICANCE: The challenges presented by the current treatment of paracoccidioidomycosis often contributing to patients' withdrawal from treatment, leading to sequelae or even death. Thus, the search for new treatment options against this disease is growing. The discovery that curcumin is active against Paracoccidioides was previously reported by our study group. Here, we clarify how the compound acts on the fungus causing its growth inhibition and decreased viability. Understanding the mechanisms of action of curcumin on P. brasiliensis elucidates how we can seek new alternatives and which metabolic pathways and molecular targets we should focus on in this incessant search to bring the patient a treatment with fewer adverse effects.
巴西副球孢子菌是副球孢子菌病的病原体，这种疾病会导致皮肤损伤，并影响肺部和其他器官。目前这种疾病的治疗时间长，且有多种副作用，这常常导致患者放弃治疗，出现后遗症，甚至死亡。因此，寻找副作用最小的新治疗形式非常重要。天然化合物是一个非常有吸引力的目标。姜黄素是姜黄根茎的主要成分之一，其药用功效在文献中有详细描述，包括对巴西副球孢子菌的抗真菌作用。然而，关于该化合物的抗真菌作用机制尚不清楚，因此本研究的目的是了解姜黄素暴露后巴西副球孢子菌的代谢发生了哪些变化，并确定该化合物的主要靶标。基于纳升超高效液相色谱-串联质谱分析（nanoUPLC-MS）的蛋白质组学分析和鉴定蛋白的功能分类。通过荧光显微镜、电子顺磁共振（EPR）和吞噬作用等生物测定方法对主要的代谢过程进行了生物学验证。蛋白质组学分析表明，姜黄素调节真菌的几种代谢过程，包括糖酵解途径、β氧化和乙醛酸循环等重要的能量产生途径。暴露于姜黄素的真菌中蛋白质合成受到下调。电子传递链和三羧酸循环也受到下调，表明线粒体膜和线粒体活性都受到了损害。暴露于该化合物后，细胞膜和细胞壁结构发生改变。真菌逃避肺泡巨噬细胞吞噬作用的能力降低。因此，姜黄素干扰引起副球孢子菌病的真菌中的几种代谢途径。生物学意义：目前副球孢子菌病的治疗方法带来的挑战常常导致患者停止治疗，导致后遗症甚至死亡。因此，针对这种疾病的新治疗方法的研究不断增加。我们研究小组之前曾报道过姜黄素对巴西副球孢子菌具有活性。在这里，我们阐明了该化合物如何作用于真菌，导致其生长抑制和活力降低。了解姜黄素对巴西副球孢子菌的作用机制阐明了我们如何寻求新的替代方法，以及在寻找新的替代方法时，我们应该关注哪些代谢途径和分子靶标。
