背景:利什曼病,由利什曼原虫属的原生动物寄生虫引起,是一种被忽视的热带病,每年有70万至100万全球新病例。与费用相关的不利影响,长期治疗和耐药性使常规治疗不利,鼓励寻找基于植物产品的替代药物。在这项研究中,在体外评估了Calotropisprocera(Asclepiadaceae)提取物对主要利什曼原虫的前鞭毛和amastigotes活力的影响。
方法:使用甲醇浸渍法制备C.procera幼苗叶片的提取物。比色细胞活力3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物(MTT)测定用于确定提取物对前鞭毛虫的生长抑制作用。在使用2''的提取物处理后,确定前精子培养物中的活性氧(ROS)水平,7'-二氯荧光素二乙酸酯(DCFH-DA)方法,并与未处理的培养物(对照)进行比较。暴露于提取物后,肿瘤坏死因子-α(TNF-α)的表达水平,确定了干扰素γ(IFN-γ)和诱导型一氧化氮合酶(iNOS)基因,并将其与感染L.major的外周血单核细胞(PBMC)中的对照进行了比较。
结果:基于MTT测定,C.procera提取物显着降低了主要前鞭毛虫的增殖,24和72小时的IC50值分别为377.28和222.44μg/mL,分别(p<0.01)。用222.44和377.28μg/mLC.procera提取物处理后,与对照相比,主要前鞭毛培养物中的ROS产量增加了1.2至1.65倍和2至4倍,分别为(p<0.05)。C.procera提取物诱导TNF-α基因表达的显着增加(2.76-14.83倍),与对照相比,感染的PBMC中的IFN-γ(25.63-三倍)和iNOS(16.32-3.97倍)(p<0.01)。
结论:根据其抗利什曼原虫活性,C.procera可以被认为是潜在治疗利什曼病的有希望的新植物来源。
BACKGROUND: Leishmaniasis, caused by protozoan parasites of the genus Leishmania, is a neglected tropical disease with 700,000 to 1,000,000 global new cases annually. Adverse effects associated with expense, long-term treatment and drug resistance have made conventional therapies unfavorable, encouraging the search for alternative drugs based on plant products. In this study, the effect of Calotropis procera (Asclepiadaceae) extract against viability of promastigotes and amastigotes of Leishmania major was evaluated in vitro.
METHODS: The extract from the leaves of C. procera seedlings was prepared using a methanol maceration method. The colorimetric cell viability 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the growth-inhibitory effect of the extract on promastigotes. The level of reactive oxygen species (ROS) in promastigote cultures was determined after treatment with the extract using the 2\',7\'-dichlorofluorescein diacetate (DCFH-DA) method and compared with untreated cultures (control). After exposure to the extract the expression levels of tumor necrosis factor-α (TNF-α), interferon gamma (IFN-γ) and inducible nitric oxide synthase (iNOS) genes were determined and compared to control in peripheral blood mononuclear cells (PBMCs) infected with L. major.
RESULTS: Based on the MTT assay, the C. procera extract significantly reduced the proliferation of L. major promastigotes with IC50 values of 377.28 and 222.44 μg/mL for 24 and 72 h, respectively (p < 0.01). After treatment with 222.44 and 377.28 μg/mL of C. procera extract, ROS production in L. major promastigote cultures increased 1.2- to 1.65-fold and 2- to 4-fold compared to the control, respectively (p < 0.05). C. procera extract induced significant increases in gene expression of TNF-α (2.76-14.83 fold), IFN-γ (25.63-threefold) and iNOS (16.32-3.97 fold) in infected PBMCs compared to control (p < 0.01).
CONCLUSIONS: On the basis of its anti-leishmanial activity, C. procera can be considered as a promising new plant source for the potential treatment of leishmaniasis.