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白花菜甲醇种子提取物的毒理学特征分析:体外和体内分析。

Toxicological profiling of methanolic seed extract of Abutilon indicum (L.) Sweet: in-vitro and in-vivo analysis.

机构信息

Immunology and Microbiology Laboratory, Department of Zoology, University of North Bengal, Darjeeling, West Bengal, 734013, India.

Genetics and Moleular Biology Labratoty, Department of Zoology, University of North Bengal, Darjeeling, West Bengal, 734013, India.

出版信息

J Ethnopharmacol. 2024 Dec 5;335:118655. doi: 10.1016/j.jep.2024.118655. Epub 2024 Aug 2.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Abutilon indicum, a shrub of the Malvaceae family, is found abundantly in tropical countries like India. A. indicum is widely used for its high medicinal properties. Traditionally, A. indicum seed powder is consumed to treat piles, constipation, chronic cystitis, gonorrhea, gleet, and pregnancy-related problems. Despite having numerous medicinal properties and widespread traditional use of A. indicum seeds, scientific validation, and toxicity studies have yet to be documented.

AIMS OF THE STUDY

The primary objective of this study is to conduct a comprehensive study on phytochemical profiling, in-vitro cytotoxicity, mutagenicity, and in-vivo acute and sub-acute toxicity, and genotoxicity on animal models of methanolic extract of A. indicum seed (MAS).

MATERIALS AND METHODS

The qualitative analysis of MAS was explored through FTIR and HR LC-MS. For in-vitro cytotoxicity, the HEK-293 cell line was used, and the TA100 (Staphylococcus typhimurium) bacterial strain was used for the Ames mutagenicity test. A single oral dose of 250, 500, 1000, or 2000 mg/kg body weight of MAS was given to each male and female rat for acute toxicity study and observed for 14 days for any toxicity signs. In the sub-acute toxicity study, 250, 500, or 1000 mg/kg body weight of MAS was administered orally to each rat for 28 days. The experimental animals were weighed weekly, and general behavior was monitored regularly. After 28 days of the experiment, the rats were sacrificed, and different serum biochemical, hematological, and histological analyses were performed. The blood samples of different doses of MAS were used for genotoxicity study through comet assay.

RESULTS

FTIR analysis found different functional groups, which indicated the presence of phenolics, flavonoids, and alkaloids. HR LC-MS analysis depicts several components with different biological functions. The cell cytotoxicity and Ames mutagenicity results showed minimal toxicity and mutagenicity up to a certain dose. The acute toxicity study conducted in Wistar albino rats demonstrated zero mortality among the animals, and the LD value for seed extract was determined to be 2000 mg/kg body weight. Sub-acute toxicity assessments indicated that the administration of seed extract resulted in no adverse effects at dosages of 250 and 500 mg/kg body weight. However, at higher doses, specifically 1000 mg/kg body weight, the liver of the experimental rats exhibited some toxic effects. In the genotoxicity study, minimal DNA damage was found in 250 and 500 mg/kg doses, respectively, but slightly greater DNA damage was found in 1000 mg/kg doses in both male and female rats.

CONCLUSIONS

The consumption of A. indicum seed powder is deemed safe; however, doses exceeding 500 mg/kg body weight may raise concerns regarding use. These findings pave the path for the creation of innovative medicines with improved efficacy and safety profiles.

摘要

民族药理学相关性

锦葵科灌木苘麻在印度等热带国家大量存在。苘麻因其具有很高的药用特性而被广泛使用。传统上,苘麻子粉被用于治疗痔疮、便秘、慢性膀胱炎、淋病、淋菌性尿道炎和与妊娠相关的问题。尽管苘麻具有许多药用特性和广泛的传统用途,但尚未对其进行科学验证和毒性研究。

研究目的

本研究的主要目的是对苘麻种子甲醇提取物(MAS)进行全面的植物化学特征分析、体外细胞毒性、致突变性、体内急性和亚急性毒性以及动物模型的遗传毒性研究。

材料和方法

采用傅里叶变换红外光谱(FTIR)和高分辨液质联用(HR LC-MS)对 MAS 进行定性分析。采用 HEK-293 细胞系进行体外细胞毒性试验,采用伤寒沙门氏菌(Staphylococcus typhimurium)TA100 菌株进行 Ames 致突变试验。雄性和雌性大鼠分别给予 MAS 250、500、1000 或 2000mg/kg 体重的单次口服剂量,观察 14 天有无毒性症状。在亚急性毒性研究中,雄性和雌性大鼠分别给予 MAS 250、500 或 1000mg/kg 体重,连续给药 28 天。每周对实验动物称重,定期监测一般行为。实验结束后 28 天处死大鼠,进行不同的血清生化、血液学和组织学分析。用彗星试验检测不同剂量 MAS 的血液样本的遗传毒性。

结果

FTIR 分析发现不同的功能团,表明存在酚类、类黄酮和生物碱。HR LC-MS 分析表明有几种具有不同生物学功能的成分。细胞毒性和 Ames 致突变性结果表明,在一定剂量下,MAS 表现出最小的毒性和致突变性。在 Wistar 白化大鼠中进行的急性毒性研究表明,动物无死亡,种子提取物的 LD 值为 2000mg/kg 体重。亚急性毒性评估表明,在 250 和 500mg/kg 体重剂量下,种子提取物的给药没有不良影响,但在更高剂量(1000mg/kg 体重)下,实验大鼠的肝脏出现一些毒性作用。在遗传毒性研究中,分别在 250 和 500mg/kg 剂量下发现最小的 DNA 损伤,但在雌雄大鼠的 1000mg/kg 剂量下发现稍大的 DNA 损伤。

结论

食用苘麻子粉被认为是安全的;然而,超过 500mg/kg 体重的剂量可能会引起对使用的担忧。这些发现为开发具有更好疗效和安全性的创新药物铺平了道路。

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