OBJECTIVE: 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).
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 LD50 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.
目的:本研究的主要目的是对植物化学剖面进行全面研究,体外细胞毒性,致突变性,以及体内急性和亚急性毒性,和遗传毒性的动物模型。
方法:通过FTIR和HRLC-MS探索MAS的定性分析。对于体外细胞毒性,使用HEK-293细胞系,并将TA100(鼠伤寒葡萄球菌)菌株用于AMES致突变性试验。在雄性和雌性大鼠中治疗250、500、1000和2000mg/kg体重的MAS的单次口服剂量以进行急性毒性研究,并观察14天的任何毒性迹象。在亚急性毒性研究中,口服给予250、500和1000mg/kg体重28天。实验动物每周称重,并定期监测一般行为。实验28天后,老鼠被处死,和不同的血清生化,血液学,并进行组织学分析。不同剂量MAS的血样通过彗星试验用于遗传毒性研究。
结果:FTIR分析发现不同的官能团,这表明酚类物质的存在,黄酮类化合物,和生物碱。HRLC-MS分析描绘了具有不同生物学功能的几种组分。细胞毒性和AMES致突变性结果显示在一定剂量下毒性和致突变性最小。在Wistar白化病大鼠中进行的急性毒性研究表明,动物的死亡率为零,种子提取物的LD50值测定为2000mg/kg体重。亚急性毒性评估表明,在250和500mg/kg体重的剂量下,施用种子提取物没有导致不良反应。然而,在更高的剂量下,特别是1000毫克/千克体重,肝脏表现出一些毒性作用。在基因毒性研究中,在250和500毫克/千克剂量中发现了最小的DNA损伤,分别,但是在雄性和雌性大鼠中,在1000mg/kg剂量中发现了更大的DNA损伤。
结论:食用A种子粉被认为是安全的;但是,剂量超过500mg/kg体重可能会引起有关使用的担忧。这些发现为创新药物的开发铺平了道路,提高了疗效和安全性。