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绒毛苹婆树皮甲醇提取物的抗利什曼原虫效力、毒性及植物化学成分评估

Evaluation of the antileishmanial potency, toxicity and phytochemical constituents of methanol bark extract of Sterculia villosa.

作者信息

Das Antu, Das Manash C, Das Niranjan, Bhattacharjee Surajit

机构信息

a Department of Molecular Biology & Bioinformatics , Tripura University (A Central University) , Suryamaninagar , India.

b Department of Chemistry , Netaji Shubhas Mahavidyalaya , Udaipur , India.

出版信息

Pharm Biol. 2017 Dec;55(1):998-1009. doi: 10.1080/13880209.2017.1285946.

Abstract

CONTEXT

Visceral leishmaniasis is a protozoan disease caused by Leishmania donovani parasite. The genus Sterculia (Malvaceae) possesses ethnobotanical potential against this protozoan infection.

OBJECTIVE

Determining the potential role of methanol bark extracts from Sterculia villosa Roxb (SVE) and its phytoconstituents against Leishmania donovani promastigotes.

MATERIALS AND METHODS

SVE was analysed by TLC, UV-Vis, IR spectroscopy and biochemical assays. Antileishmanial potential of SVE (0.5-130 μg/mL for 72 h) was characterized by MTT assay. Fluorescent microscopy was performed to validate the IC dose. To determine the effect of SVE on promastigotes, reactive oxygen species (ROS) and superoxide generation, lipid peroxidation and DNA fragmentation assays were performed. Molecular aggregation of compounds was determined by atomic force microscopy (AFM). Extent of cytotoxicity of SVE at IC dose was determined against RAW 264.7 macrophages, peritoneal macrophages and murine RBCs. In vivo cytotoxicity of SVE was evaluated in BALB/c mice.

RESULT

SVE exhibited reverse dose dependent antileishmanial activity when 130-0 μg/mL doses were tested against promastigotes. The IC and IC values were found to be 17.5 and 10 μg/mL, respectively. SVE at IC dose demonstrated elevated level of ROS, superoxide, lipid peroxidation and DNA fragmentation against promastigotes with no cytotoxicity. AFM analysis suggested increasing size of molecular aggregation (31.3 nm < 35.2 nm < 2.93 μm) with increase in concentration (10 μg < 17.5 μg < 130 μg).

DISCUSSION AND CONCLUSIONS

The study elucidates the antileishmanial potential of SVE against Leishmania donovani promastigotes by exerting oxidative stress and DNA damage. In sum, SVE can be explored as an immunotherapeutic candidate against leishmaniasis and other infectious diseases.

摘要

背景

内脏利什曼病是由杜氏利什曼原虫寄生虫引起的原生动物疾病。苹婆属(锦葵科)具有针对这种原生动物感染的民族植物学潜力。

目的

确定绒毛苹婆(SVE)树皮甲醇提取物及其植物成分对杜氏利什曼原虫前鞭毛体的潜在作用。

材料与方法

通过薄层色谱法、紫外可见光谱法、红外光谱法和生化分析对SVE进行分析。通过MTT法表征SVE(0.5 - 130μg/mL,作用72小时)的抗利什曼原虫潜力。进行荧光显微镜检查以验证半数抑制浓度(IC)剂量。为了确定SVE对前鞭毛体的影响,进行了活性氧(ROS)和超氧化物生成、脂质过氧化和DNA片段化测定。通过原子力显微镜(AFM)确定化合物的分子聚集情况。在IC剂量下,针对RAW 264.7巨噬细胞、腹腔巨噬细胞和小鼠红细胞测定SVE的细胞毒性程度。在BALB/c小鼠中评估SVE的体内细胞毒性。

结果

当以130 - 0μg/mL剂量对前鞭毛体进行测试时,SVE表现出反向剂量依赖性抗利什曼原虫活性。发现IC和IC值分别为17.5和10μg/mL。在IC剂量下,SVE对前鞭毛体表现出升高的ROS、超氧化物、脂质过氧化和DNA片段化水平,且无细胞毒性。AFM分析表明,随着浓度增加(10μg < 17.5μg < 130μg),分子聚集尺寸增大(31.3nm < 35.2nm < 2.93μm)。

讨论与结论

该研究阐明了SVE通过施加氧化应激和DNA损伤对杜氏利什曼原虫前鞭毛体的抗利什曼原虫潜力。总之,SVE可作为抗利什曼病和其他传染病的免疫治疗候选药物进行探索。

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