Doctoral Program in Biomedical Sciences, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia.
Department of Medical Laboratory Technology, Poltekkes Kemenkes Kupang, Kupang, Indonesia.
Open Vet J. 2024 Sep;14(9):2294-2309. doi: 10.5455/OVJ.2024.v14.i9.18. Epub 2024 Sep 30.
Protein deficiency poses a significant challenge during the growth and development of children. Lam (MO) leaves, renowned for their high protein content, present a potential solution to address amino acid imbalances in protein-deficient conditions.
This study aimed to evaluate the toxicity of MO leaf extract in Sprague-Dawley Rats.
Protein extraction from (MO) leaves was performed using ultrasonic-assisted extraction (UAE) with ethanol. The ethanol leaf extract of MO (EEMO) was then characterized for protein content, amino acids, minerals, phytic acid, and phytochemicals. Antioxidant activity was assessed using the DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) test, while cytotoxicity was evaluated using the MTT (3-(4,5-dimethylthiazolyl-2)-2,5 diphenyltetrazolium bromide) assay on HepG2 and Madin-Darby canine kidney (MDCK) cells. EEMO was tested for acute toxicity in 60 healthy male and female Sprague-Dawley rats. The rats were divided into five groups of five rats each, receiving single oral doses of 5, 50, 300, 2,000, and 5,000 mg/kg body weight (BW) of EEMO. Observations were conducted daily till day 14 before the necropsy of rats. Liver and kidney tissues were harvested and preserved in 10% formalin for histopathological analysis.
The extraction process revealed a protein content of 45.5%, with phenylalanine being the predominant essential amino acid at 22.25 mg/g, and glutamic acid as the dominant non-essential amino acid at 60.03 mg/g. Potassium (1174.23 mg/100 g) and selenium (149 mg/100 g) were identified as the primary macro and micro minerals, respectively. The IC and CC values for antioxidant and cytotoxic activities in HepG2 and MDCK were found to be 41.04,182.66, and 121.04 ppm, respectively. Toxicity testing on experimental animals resulted in an LD value of 5,000 mg/kg BW for EEMO, indicating its relative safety upon oral administration.
MO extract produced by the UAE extraction method, containing high-quality food-grade protein, showed no cytotoxic effects on HepG2 and MDCK cells and exhibited no acute toxicity in rats.
蛋白质缺乏是儿童生长发育过程中的重大挑战。由于Lam（MO）树叶蛋白质含量高，因此成为解决蛋白质缺乏条件下氨基酸失衡问题的潜在解决方案。
本研究旨在评估 MO 叶提取物在 Sprague-Dawley 大鼠中的毒性。
采用超声辅助提取（UAE）乙醇从 MO 叶中提取蛋白质。然后对 MO 的乙醇叶提取物（EEMO）进行了蛋白质含量、氨基酸、矿物质、植酸和植物化学物质的表征。使用 DPPH（2,2-二苯基-1-苦基肼水合物）试验评估抗氧化活性，使用 MTT（3-（4,5-二甲基噻唑-2）-2,5-二苯基四唑溴化物）试验评估 HepG2 和 Madin-Darby 犬肾（MDCK）细胞的细胞毒性。对 60 只健康雄性和雌性 Sprague-Dawley 大鼠进行了 EEMO 的急性毒性试验。大鼠分为五组，每组 5 只，分别口服 5、50、300、2000 和 5000 mg/kg 体重（BW）的 EEMO。每天观察大鼠至第 14 天，然后进行解剖。采集肝脏和肾脏组织，用 10%福尔马林保存，用于组织病理学分析。
提取过程显示蛋白质含量为 45.5%，苯丙氨酸为 22.25mg/g，是主要的必需氨基酸，谷氨酸为 60.03mg/g，是主要的非必需氨基酸。钾（1174.23mg/100g）和硒（149mg/100g）分别是主要的宏量和微量元素。在 HepG2 和 MDCK 中，抗氧化和细胞毒性活性的 IC 和 CC 值分别为 41.04、182.66 和 121.04ppm。对实验动物的毒性试验表明，EEMO 的 LD 值为 5000mg/kg BW，表明其口服给药相对安全。
采用 UAE 提取方法提取的 MO 提取物，含有高质量的食品级蛋白质，对 HepG2 和 MDCK 细胞无细胞毒性作用，在大鼠中无急性毒性。
