在体外条件下,通过增加 ROS 的产生和上调 TNF-α、IFN-γ 和 iNOS mRNA 的表达,提高草蒌提取物的抗利什曼原虫活性。

IF 3.6 Q1 TROPICAL MEDICINE Tropical Medicine and Health Pub Date : 2024-02-01 DOI:10.1186/s41182-024-00578-4
Shahla Amani, Soheila Alinejad, Negar Asadi, Elham Yousefi, Shahram Khademvatan, Gordon Stanley Howarth
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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). 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引用次数: 0

摘要

背景:利什曼病是由利什曼属原生动物寄生虫引起的,是一种被忽视的热带疾病,全球每年新增病例 70 万至 100 万。与费用、长期治疗和耐药性相关的不良反应使传统疗法难以奏效,因此人们开始寻找基于植物产品的替代药物。在这项研究中,我们在体外评估了草木犀(Asclepiadaceae)提取物对大利什曼原虫和非原虫活力的影响:方法:采用甲醇浸泡法从 C. procera 幼苗的叶片中提取提取物。采用比色细胞活力 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑(MTT)测定法确定萃取物对原原体的生长抑制作用。用 2',7'-二氯荧光素二乙酸酯(DCFH-DA)法测定原生动物培养物中活性氧(ROS)的水平,并与未处理的培养物(对照组)进行比较。接触提取物后,测定了感染大肠杆菌的外周血单核细胞(PBMC)中肿瘤坏死因子-α(TNF-α)、γ干扰素(IFN-γ)和诱导型一氧化氮合酶(iNOS)基因的表达水平,并与对照组进行了比较:结果:根据 MTT 试验,C.procera 提取物能显著减少大鼠原虫的增殖,24 小时和 72 小时的 IC50 值分别为 377.28 和 222.44 μg/mL(p 结论:C.procera 提取物能显著减少大鼠原虫的增殖,24 小时和 72 小时的 IC50 值分别为 377.28 和 222.44 μg/mL:基于其抗利什曼病活性,C. procera 可被视为一种有潜力治疗利什曼病的新植物来源。
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Anti-Leishmania major activity of Calotropis procera extract by increasing ROS production and upregulating TNF-α, IFN-γ and iNOS mRNA expression under in vitro conditions.

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.

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来源期刊
Tropical Medicine and Health
Tropical Medicine and Health TROPICAL MEDICINE-
CiteScore
7.00
自引率
2.20%
发文量
90
审稿时长
11 weeks
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