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白桑(Morus alba linn.)乙醇叶提取物对雌性小鼠的生物活性、植物化学研究及亚急性体内毒性

Bioactivity, phytochemistry studies and subacute in vivo toxicity of ethanolic leaf extract of white mulberry (Morus alba linn.) in female mice.

作者信息

Fauzi Ahmad, Kifli Nurolaini, Noor Mohd Hezmee Mohd, Hamzah Hazilawati, Azlan Azrina

机构信息

Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia UPM, Serdang, 43400, Selangor, Malaysia; Department of Clinical Pathology, Faculty of Veterinary Medicine, University of Brawijaya, Malang, East Java, 65141, Indonesia.

PAP Rashidah Sa'adatul Bolkiah Institute of Health Sciences, Universiti Brunei Darussalam, Gadong, BE, 1410, Brunei.

出版信息

J Ethnopharmacol. 2024 May 10;325:117914. doi: 10.1016/j.jep.2024.117914. Epub 2024 Feb 13.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Traditional uses of Morus alba L. leaf extracts (MLE) have been reported for treating hyperglycaemia and diabetes. Phytochemical compounds in the leaves demonstrated the ability to enhance insulin sensitivity and β-cell secretory function, suggesting their potential value in reducing blood glucose and treating diabetes. However, the phytochemical constituents and safety of the herbal medicines need to be verified in each experimental field from different growing areas. Studies on the phytochemistry and toxicity of Morus alba leaves in Southeast Asia, especially in Brunei, have never been investigated.

AIM OF THE STUDY

This study aimed to investigate the bioactivity and phytochemistry of Morus alba ethanolic leaf extract from Brunei Darussalam and its subacute toxic effects in the Institute of Cancer Research (ICR) female mice.

MATERIALS AND METHODS

The phenolic yield and antioxidant of the extract were analysed. Meanwhile, liquid chromatography-mass spectrometry and high-performance liquid chromatography were utilised to determine the phenolic compound of the MLE. In the subacute toxicity study, twenty-five female mice were randomly divided into five groups: the control group, which received oral gavage of 5% dimethyl sulfoxide solvent (DMSO), and the MLE treatment group, which received the extract at a dose of 125, 250, 500 and 1000 mg/kg. Physiology, haematology, biochemistry, and histology were evaluated during the study.

RESULTS

Morus alba leaf depicted total phenolic 10.93 mg gallic acid equivalents (GAE)/g dry weight (DW), flavonoid 256.67 mg quercetin equivalents (QE)/g DW, and antioxidant bioactivity content of 602.03 IC μg/mL and 13.21 mg Fe2+/g DW. Twenty compounds in the Morus alba ethanolic leaf extract were identified, with chlorogenic acid (305.60 mg/100 g DW) as the primary compound. As for subacute toxicity in this study, neither mortality nor haematological changes were observed. On the other hand, administration of 500 and 1000 mg/kg MLE resulted in mild hepatocellular injury, as indicated by a significant (p < 0.05) increase in liver enzyme activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The histopathological score showed mild hepatocellular necrosis in administering 250, 500, and 1000 mg/kg of MLE. The parameters of renal injury were within normal limits, with the increase in eosinophilic cytoplasm observed in the histological scoring at 1000 mg/kg of MLE.

CONCLUSIONS

Morus alba leaf extract showed abundant polyphenols. In a study on subacute toxicity, MLE caused mild hepatotoxicity in mice. The toxic effect of the extract may be due to kaempferol and chlorogenic acid compounds. The 125 mg/kg MLE dose was safe with no adverse effects.

摘要

民族药理学相关性

据报道,白桑叶提取物(MLE)的传统用途是治疗高血糖和糖尿病。叶片中的植物化学化合物具有增强胰岛素敏感性和β细胞分泌功能的能力,表明其在降低血糖和治疗糖尿病方面具有潜在价值。然而,草药的植物化学成分和安全性需要在来自不同种植地区的每个实验领域进行验证。东南亚,特别是文莱的白桑叶植物化学和毒性研究从未进行过。

研究目的

本研究旨在调查文莱达鲁萨兰国白桑叶乙醇提取物的生物活性、植物化学及其对癌症研究所(ICR)雌性小鼠的亚急性毒性作用。

材料与方法

分析提取物的酚类产量和抗氧化剂。同时,利用液相色谱 - 质谱联用和高效液相色谱法测定MLE的酚类化合物。在亚急性毒性研究中,25只雌性小鼠随机分为五组:对照组口服5%二甲基亚砜溶剂(DMSO),MLE治疗组分别接受125、250、500和1000mg/kg剂量的提取物。研究期间评估生理、血液学、生物化学和组织学指标。

结果

白桑叶的总酚含量为10.93mg没食子酸当量(GAE)/g干重(DW),黄酮类化合物含量为256.67mg槲皮素当量(QE)/g DW,抗氧化生物活性含量为602.03 IC μg/mL和13.21mg Fe2+/g DW。鉴定出白桑叶乙醇提取物中的20种化合物,其中绿原酸(305.60mg/100g DW)为主要化合物。在本研究的亚急性毒性方面,未观察到死亡或血液学变化。另一方面,给予500和1000mg/kg MLE导致轻度肝细胞损伤,丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)的肝酶活性显著(p < 0.05)升高表明了这一点。组织病理学评分显示,给予250、500和1000mg/kg MLE时出现轻度肝细胞坏死。肾损伤参数在正常范围内,在给予1000mg/kg MLE的组织学评分中观察到嗜酸性细胞质增加。

结论

白桑叶提取物显示出丰富的多酚。在亚急性毒性研究中,MLE对小鼠造成轻度肝毒性。提取物的毒性作用可能归因于山奈酚和绿原酸化合物。125mg/kg MLE剂量是安全的,没有不良反应。

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