Alanezi Abdulkareem A
Department of Pharmaceutics, College of Pharmacy, University of Hafr Al Batin, Hafr Al Batin 39524, Saudi Arabia.
Metabolites. 2024 Dec 23;14(12):726. doi: 10.3390/metabo14120726.
BACKGROUND/OBJECTIVES: Catha edulis, commonly known as khat, is used for its psychoactive effects and is considered a natural amphetamine. The current study investigated the metabolomic profile in the cerebellum of mice after repeated exposure to khat and evaluated the effects of clavulanic acid on the metabolomic profile in the cerebellum in khat-treated mice.
Male C67BL/6 mice that were 6-9 weeks old were recruited and divided into three groups: the control group was treated with 0.9% normal saline for 17 days; the khat group was given khat extract at a dose of 360 mg/kg via the intraperitoneal (i.p) route for 17 days; and another khat group was treated with khat for 17 days and clavulanic acid at a dose of 5 mg/kg for the last 7 days (days 11-17). At the end of the 17th day, the animals were sacrificed, and their brains were immediately collected and stored at -80 °C. The cerebellum region of the brain was isolated in each group by micropuncture using cryostat and underwent a metabolomics study via Gas Chromatography/Mass Spectroscopy (GC/MS). The total peak area ratios of the selected metabolites in the cerebellum after repeated exposure to the khat extract were significantly reduced ( < 0.05) and treatment of the khat group with clavulanic acid significantly increased (all < 0.05) the total peak areas ratios of the selected metabolites when compared to their corresponding areas in the alternative khat group. These levels of selected metabolites were further confirmed by observing the metabolite peak area ratios and performing a heat map analysis and a principal compartment analysis of the samples in the cerebellum.
A network analysis of altered metabolites in the cerebellum showed a strong correlation between the different metabolites, which showed that an increase in one metabolite can modulate the levels of others. An analysis using the MetaboAnalyst software revealed the involvement of selected altered metabolites like lactic acid in many signaling pathways, like gluconeogenesis, while enrichment analysis data showed altered pathways for pyruvate metabolism and disease pathogenesis. Finally, a network analysis showed that selected metabolites were linked with other metabolites, indicating drug-drug interactions.
The present study showed that repeated exposure of mice to khat altered the levels of various metabolites in the cerebellum which are involved in the pathogenesis of different diseases, signaling pathways, and interactions with the pharmacokinetic profile of other therapeutic drugs. The treatment of khat-treated mice with clavulanic acid positively modified the metabolomics profile in the cerebellum and increased the levels of the altered metabolites.
背景/目的：巧茶，俗称恰特草，因其具有精神活性作用而被使用，被认为是一种天然安非他命。本研究调查了小鼠反复接触恰特草后小脑的代谢组学特征，并评估了克拉维酸对恰特草处理小鼠小脑代谢组学特征的影响。
招募6 - 9周龄的雄性C67BL/6小鼠，分为三组：对照组用0.9%生理盐水处理17天；恰特草组通过腹腔注射（i.p）途径给予360 mg/kg的恰特草提取物，持续17天；另一恰特草组用恰特草处理17天，并在最后7天（第11 - 17天）给予5 mg/kg的克拉维酸。在第17天结束时，处死动物，立即收集其大脑并储存在 - 80°C。通过使用低温恒温器进行微穿刺，在每组中分离出大脑的小脑区域，并通过气相色谱/质谱联用（GC/MS）进行代谢组学研究。与另一恰特草组相比，反复接触恰特草提取物后，小脑中所选代谢物的总峰面积比显著降低（<0.05），而用克拉维酸处理恰特草组显著增加了所选代谢物的总峰面积比（均<0.05）。通过观察代谢物峰面积比并对小脑中的样本进行热图分析和主成分分析，进一步证实了所选代谢物的这些水平。
对小脑中改变的代谢物进行网络分析表明，不同代谢物之间存在强相关性，这表明一种代谢物的增加可以调节其他代谢物的水平。使用MetaboAnalyst软件进行的分析揭示了所选改变的代谢物如乳酸参与了许多信号通路，如糖异生，而富集分析数据显示丙酮酸代谢和疾病发病机制的途径发生了改变。最后，网络分析表明所选代谢物与其他代谢物相关联，表明存在药物 - 药物相互作用。
本研究表明，小鼠反复接触恰特草会改变小脑中各种代谢物的水平，这些代谢物参与不同疾病的发病机制、信号通路以及与其他治疗药物药代动力学特征的相互作用。用克拉维酸处理恰特草处理的小鼠可积极改变小脑的代谢组学特征，并增加改变的代谢物水平。
