Portela José Luiz, Bianchini Matheus Chimelo, Roos Daniel H, de Ávila Daiana Silva, Puntel Robson Luiz
Campus Uruguaiana, Programa de Pós-Graduação em Bioquímica (PPGBioq), Universidade Federal do Pampa, Campus Uruguaiana BR-472 Km 7, Uruguaiana, RS, CEP 97500-970, Brazil.
Naunyn Schmiedebergs Arch Pharmacol. 2021 Feb;394(2):227-240. doi: 10.1007/s00210-020-01974-3. Epub 2020 Sep 15.
The deficiency in the activity of the mitochondrial enzyme methylmalonyl-CoA mutase (MCM, EC 5.4.99.2) leads to a condition called methylmalonic academia, which is characterised by the accumulation of methylmalonic (MMA), malonic (MA) or other organic acids. Importantly, we have recently found that supplementation with Ilex paraguariensis aqueous extract offered protection against toxicity associated with MMA or MA exposure to Drosophila melanogaster. Of note, caffeic acid (CA) and caffeine (CAF) were the major phytochemicals found in our Ilex paraguariensis crude extract. Therefore, here, we have exploited CA and/or CAF to test the hypothesis that supplementation with the isolated compounds (either alone or combined) could exert a protective effect against MMA or MA-induced toxicity in flies. Therefore, flies were exposed to MA (5 mM) or MMA (5 mM) and concomitantly treated with CA (1.39 μg/mL), CAF (1.27 μg/mL) or CA + CAF for 10 days for survival, and for 4 days for behavioural and biochemical assays. CA, CAF and CA + CAF treatments completely abolished the mortality associated with either MMA or MA exposure. Moreover, CA and CAF, either alone or combined, completely abolished behavioural changes, and completely protect against changes in thiobarbituric acid reactive substances (TBARS) levels, catalase (CAT) activity and MTT reduction ability, associated with MA or MMA exposure. In turn, CAF restored SOD activity in the head of flies exposed to MA or MMA. However, CA and CAF (either alone or combined) significantly decreased acetylcholinesterase (AChE) activity per se, while CAF alone protected from changes in AChE activity (in head tissue) associated with MA or MMA. Finally, CA and/or CAF were able to protect from a decrease in glucose and triglyceride levels associated with both MA and MMA exposures in haemolymph. Together, our data confirm the hypothesis that supplementation with CA and/or CAF offers protection against detrimental changes associated with MMA or MA exposure in flies, being responsible, at least in part, for the protective effect of I. paraguariensis crude extract which was reported previously.
线粒体酶甲基丙二酰辅酶A变位酶（MCM，EC 5.4.99.2）活性的缺乏会导致一种名为甲基丙二酸血症的病症，其特征是甲基丙二酸（MMA）、丙二酸（MA）或其他有机酸的积累。重要的是，我们最近发现，补充巴拉圭冬青水提取物可提供针对与MMA或MA暴露相关的毒性的保护作用，以果蝇为实验对象。值得注意的是，咖啡酸（CA）和咖啡因（CAF）是我们在巴拉圭冬青粗提取物中发现的主要植物化学物质。因此，在这里，我们利用CA和/或CAF来检验以下假设：补充分离出的化合物（单独或组合使用）可以对果蝇中MMA或MA诱导的毒性发挥保护作用。因此，将果蝇暴露于MA（5 mM）或MMA（5 mM）中，并同时用CA（1.39 μg/mL）、CAF（1.27 μg/mL）或CA + CAF处理10天以观察存活率，处理4天以进行行为和生化分析。CA、CAF和CA + CAF处理完全消除了与MMA或MA暴露相关的死亡率。此外，CA和CAF单独或组合使用，完全消除了行为变化，并完全防止了与MA或MMA暴露相关的硫代巴比妥酸反应性物质（TBARS）水平、过氧化氢酶（CAT）活性和MTT还原能力的变化。反过来，CAF恢复了暴露于MA或MMA的果蝇头部的超氧化物歧化酶（SOD）活性。然而，CA和CAF（单独或组合）本身显著降低了乙酰胆碱酯酶（AChE）活性，而单独的CAF则防止了与MA或MMA相关的（头部组织中的）AChE活性变化。最后，CA和/或CAF能够防止血淋巴中与MA和MMA暴露相关的葡萄糖和甘油三酯水平降低。总之，我们的数据证实了以下假设：补充CA和/或CAF可针对果蝇中与MMA或MA暴露相关的有害变化提供保护，这至少部分地解释了先前报道的巴拉圭冬青粗提取物的保护作用。
