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通过气相色谱-质谱联用仪对脂肪酸进行表征、在多个阶段的抗糖化作用以及对摩洛哥东部种植的仙人掌(Opuntia ficus-indica (L.) Mill)籽油白蛋白糖基化诱导的红细胞氧化损伤的保护作用:实验和计算方法。

Fatty acids characterisation by GC-MS, antiglycation effect at multiple stages and protection of erythrocytes cells from oxidative damage induced by glycation of albumin of Opuntia ficus-indica (L.) Mill seed oil cultivated in Eastern Morocco: Experimental and computational approaches.

作者信息

Abdnim Rhizlan, Lafdil Fatima Zahra, Elrherabi Amal, El Fadili Mohamed, Kandsi Fahd, Benayad Ouijdane, Legssyer Abdelkhaleq, Ziyyat Abderrahim, Mekhfi Hassane, Bnouham Mohamed

机构信息

Laboratory of Bioresources, Biotechnology, Ethnopharmacology, and Health, Faculty of Sciences, Mohammed First University, Oujda, B.P. 717, Morocco.

Laboratory of Bioresources, Biotechnology, Ethnopharmacology, and Health, Faculty of Sciences, Mohammed First University, Oujda, B.P. 717, Morocco.

出版信息

J Ethnopharmacol. 2024 Jul 15;329:118106. doi: 10.1016/j.jep.2024.118106. Epub 2024 Apr 2.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Opuntia ficus-indica (L.) Mill is frequently observed in the Moroccan traditional medicinal system, where these approaches are employed to mitigate the onset of diabetes and the subsequent complications it may entail.

AIM OF THE STUDY

The aim of this research was to examine the effectiveness of Opuntia ficus-indica seed oil in preventing diabetic complications. Specifically, the study assessed its ability to counteract glycation at various stages, protected red blood cells from the harmful effects of glycated albumin, and inhibited pancreatic lipase digestive enzymes to understand its potential antihyperglycemic properties. Additionally, the study aimed to identify the chemical components responsible for these effects, evaluate antioxidant and anti-inflammatory properties, and conduct computational investigations such as molecular docking.

MATERIALS AND METHODS

The assessement of Opuntia ficus-indica seed oil antiglycation properties involved co-incubating the extract oil with a bovine serum albumin-glucose glycation model. The study investigated various stages of glycation, incorporating fructosamine (inceptive stage), protein carbonyls (intermediate stage), and AGEs (late stage). Additionally, measurement of β-amyloid aggregation of albumin was performed using Congo red, which is specific to amyloid structures. Additionally, the evaluation of oil's safeguarding effect on erythrocytes against toxicity induced by glycated albumin included the measurement of erythrocyte hemolysis, lipid peroxidation, reduced glutathione. The fatty acid of Opuntia ficus-indica seed oil were analyzed using Gas Chromatography-Mass Spectrometry (GC-MS). The in vitro evaluation of antihyperglycemic activity involved the use of pancreatic lipase enzyme, while the assessement of antioxidant capability was carried out through the utilization of the ABTS and FRAP methods. The in vitro assessement of the denaturation of albumin activity was also conducted. In conjunction with the experimental outcomes, computational investigations were undertaken, specifically employing ADMET (absorption, distribution, metabolism, excretion, and toxicity) analysis. Furthermore, molecular docking was utilized to predict antioxidant and antiglycation mechanisms based on protein targets.

RESULTS

In vitro glycation assays, Opuntia ficus-indica seed oil displayed targeted inhibitory effects at multiple distinct stages. Within erythrocytes, in addition to mitigating hemolysis and lipid peroxidation induced by glycated albumin. GC-MS investigation revealed a richness of fatty acids and the most abundant compounds are Linoleic acid (36.59%), Palmitic acid (20.84%) and Oleic acid (19.33%) respectively. The findings of antioxidant ability showed a remarkable activity on FRAP and ABTS radicals. This oil showed a pronounced inhibitory impact (p < 0.001) on pancreatic lipase enzyme. It also exerted a notibale inhibition of albumin denaturation, in vitro.

CONCLUSION

The identified results were supported by the abundant compounds of fatty acids unveiled through GC-MS analysis, along with the computational investigation and molecular docking.

摘要

民族药理学相关性

在摩洛哥传统医学体系中经常能见到印度仙人掌(Opuntia ficus-indica (L.) Mill),人们采用这些方法来减轻糖尿病的发病以及可能随之而来的并发症。

研究目的

本研究旨在检验印度仙人掌籽油预防糖尿病并发症的有效性。具体而言,该研究评估了其在不同阶段对抗糖基化的能力、保护红细胞免受糖化白蛋白有害影响的能力以及抑制胰腺脂肪酶消化酶的能力,以了解其潜在的降血糖特性。此外,该研究旨在确定产生这些作用的化学成分,评估抗氧化和抗炎特性,并进行诸如分子对接等计算研究。

材料与方法

对印度仙人掌籽油抗糖基化特性的评估涉及将提取的油与牛血清白蛋白 - 葡萄糖糖基化模型共同孵育。该研究调查了糖基化的各个阶段,包括果糖胺(起始阶段)、蛋白质羰基(中间阶段)和晚期糖基化终末产物(晚期阶段)。此外,使用对淀粉样结构具有特异性的刚果红对白蛋白的β - 淀粉样蛋白聚集进行了测量。此外,评估该油对红细胞免受糖化白蛋白诱导的毒性的保护作用包括测量红细胞溶血、脂质过氧化、还原型谷胱甘肽。使用气相色谱 - 质谱联用仪(GC - MS)分析了印度仙人掌籽油的脂肪酸。体外降血糖活性评估使用了胰腺脂肪酶,而抗氧化能力评估则通过ABTS和FRAP方法进行。还进行了白蛋白活性变性的体外评估。结合实验结果,进行了计算研究,具体采用ADMET(吸收、分布、代谢、排泄和毒性)分析。此外,利用分子对接基于蛋白质靶点预测抗氧化和抗糖基化机制。

结果

在体外糖基化试验中,印度仙人掌籽油在多个不同阶段表现出靶向抑制作用。在红细胞内,除了减轻糖化白蛋白诱导的溶血和脂质过氧化外。GC - MS研究表明脂肪酸种类丰富,最丰富的化合物分别是亚油酸(36.59%)、棕榈酸(20.84%)和油酸(19.33%)。抗氧化能力的研究结果显示对FRAP和ABTS自由基具有显著活性。这种油对胰腺脂肪酶表现出显著的抑制作用(p < 0.001)。在体外,它对白蛋白变性也有明显的抑制作用。

结论

通过GC - MS分析揭示的丰富脂肪酸化合物以及计算研究和分子对接支持了所确定的结果。

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