Neris Débora Meira, Ortolani Letícia Gonçalves, de Castro Cynthia Aparecida, Correia Ricardo de Oliveira, Rodolpho Joice Margareth de Almeida, Camillo Luciana, Nogueira Camila Tita, de Sousa Cristina Paiva, Anibal Fernanda de Freitas
Laboratory of Inflammation and Infectious Diseases, Department of Morphology and Pathology, Federal University of São Carlos, São Carlos, Brazil.
Laboratory of Microbiology and Biomolecules, Federal University of São Carlos, São Carlos, Brazil.
Int J Microbiol. 2020 Aug 27;2020:8895308. doi: 10.1155/2020/8895308. eCollection 2020.
Leishmaniases are diseases with high epidemiological relevance and wide geographical distribution. In Brazil, is related to the tegumentary form of leishmaniasis. The treatment for those diseases is problematic as the available drugs promote adverse effects in patients. Therefore, it is important to find new therapeutic targets. In this regard, one alternative is the study of biomolecules produced by endophytic microorganisms. In this study, the total extract produced by the endophytic RNC-D was used to evaluate the leishmanicidal, nitric oxide, and cytokines production using RAW 264.7 macrophages. The results showed that, in the leishmanicidal assay with , EC50 values at the periods of 24 and 48 hours were 0.624 mg/mL and 0.547 mg/mL, respectively. Furthermore, the cells treated with the extract presented approximately 25% of infected cells with an average of 3 amastigotes/cell in the periods of 24 and 48 hours. Regarding the production of cytokines in RAW 264.7 macrophages infected/treated with the extract, a significant increase in TNF- was observed at the periods of 24 and 48 hours in the treated cells. The concentrations of IFN- and IL-12 showed significant increase in 48 hours. A significant decrease in IL-4 was observed in all cells treated with the extract in 24 hours. It was observed in the treated cells that the NO production by RAW 264.7 macrophages increased between 48 and 72 hours. The endophytic RNC-D extract modulates the mediators of inflammation produced by RAW 264.7 macrophages promoting death. The immunomodulatory effects might be a promising target to develop new immunotherapeutic and antileishmanial drugs.
利什曼病是具有高度流行病学相关性且地理分布广泛的疾病。在巴西,它与皮肤利什曼病有关。这些疾病的治疗存在问题,因为现有药物会对患者产生不良反应。因此,寻找新的治疗靶点很重要。在这方面,一种选择是研究内生微生物产生的生物分子。在本研究中,内生真菌RNC-D产生的总提取物被用于使用RAW 264.7巨噬细胞评估其杀利什曼原虫、一氧化氮和细胞因子的产生。结果表明,在使用该提取物的杀利什曼原虫试验中,24小时和48小时的半数有效浓度(EC50)值分别为0.624毫克/毫升和0.547毫克/毫升。此外,在24小时和48小时期间,用提取物处理的细胞中约25%的感染细胞平均每个细胞有3个无鞭毛体。关于用提取物感染/处理的RAW 264.7巨噬细胞中细胞因子的产生,在处理后的细胞中,24小时和48小时时肿瘤坏死因子-α(TNF-α)有显著增加。干扰素-γ(IFN-γ)和白细胞介素-12(IL-12)的浓度在48小时时显著增加。在24小时时,用提取物处理的所有细胞中白细胞介素-4(IL-4)有显著下降。在处理后的细胞中观察到,RAW 264.7巨噬细胞产生的一氧化氮在48小时至72小时之间增加。内生真菌RNC-D提取物调节RAW 264.7巨噬细胞产生的炎症介质,促进利什曼原虫死亡。这种免疫调节作用可能是开发新的免疫治疗和抗利什曼病药物的一个有前景的靶点。