Silva Tuani Mendes, Fracasso Débora Soligo, Vargas Visentin Ana Paula, Cassini Carina, Scariot Fernando Joel, Danetti Sidineia, Echeverrigaray Sergio, Moura Sidnei, Touguinha Luciana Bavaresco, Branco Catia Santos, Salvador Mirian
Laboratório de Estresse Oxidativo e Antioxidantes, Instituto de Biotecnologia, Universidade de Caxias Do Sul. Rua Francisco Getúlio Vargas, 1130, Caxias Do Sul, Rio Grande do Sul, CEP: 95070-560, Brazil.
Laboratório de Enologia e Microbiologia Aplicada, Instituto de Biotecnologia, Universidade de Caxias Do Sul. Rua Francisco Getúlio Vargas, 1130, Caxias Do Sul, Rio Grande do Sul, CEP: 95070-560, Brazil.
J Ethnopharmacol. 2022 Nov 15;298:115564. doi: 10.1016/j.jep.2022.115564. Epub 2022 Aug 6.
Matcha green tea (Camellia sinensis) based-supplements have been widely used since they present a greater content of phenolic compounds than traditional green tea, which is popularly used in the treatment of diabetes. However, there are few studies on the effectiveness and safety of matcha supplements.
This work aimed to evaluate the efficacy and safety of this supplement in endothelial cells (EA.hy926) in the hyperglycemic model and in vivo Artemia salina.
To assess the effect of Matcha herbal supplement (MHS), EA. hy926 endothelial cells were treated with 20 μg/mL of MHS for 24 h, in a hyperglycemic medium with 35 mM glucose. After treatment, cells were trypsinized and centrifuged at 4 °C and 47×g for 5 min. The pellet was used to determine the reaction products to thiobarbituric acid and the levels of nitric oxide. Electron transport chain activity and ATP levels were also evaluated. Intracellular pH, apoptosis, and mitochondrial membrane depolarization were evaluated by flow cytometry. MHS chemical characterization was performed by HPLC-UV and total phenolic content analysis. The evaluation of the antioxidant capacity of MHS was performed by 2,2-diphenyl-1-picrylhydrazyl radical scavenger assay. To determine the in vivo acute toxicity of MHS, an A. salina assay was conducted, using 0,2 mL of different concentrations of MHS (10, 50, 100, 250, 500, 750 and 1000 μg/mL). The LD values were obtained by interpolation of 50% (y = 50) of the dead individuals in the trend curves.
Our data showed that MHS was able to avoid oxidative and nitrosative stress induced by hyperglycemia, demonstrating important antioxidant activity. However, it was observed that MHS reduced up to 90% the activity of the four-electron transport complexes, reducing the ATP production of the endothelial cells. In the toxicity assay performed in Artemia salina, MHS showed mild toxicity (LD = 0,4 mg/mL). The major compounds found in MHS were epigallocatechin gallate, epicatechin, rutin, kaempferol, and quercetin.
This data draws attention to the fact that supplements with high content of phenolic compounds, capable of avoiding oxidative and nitrosative stress can have a dual effect and, simultaneously to antioxidant activity, can induce toxicity in different cell types.
基于抹茶绿茶(茶树)的补充剂已被广泛使用,因为它们比传统绿茶含有更多的酚类化合物,传统绿茶常用于治疗糖尿病。然而,关于抹茶补充剂有效性和安全性的研究很少。
这项工作旨在评估这种补充剂在高血糖模型中的内皮细胞(EA.hy926)和体内卤虫中的功效和安全性。
为了评估抹茶草药补充剂(MHS)的效果,在含有35 mM葡萄糖的高血糖培养基中,用20 μg/mL的MHS处理EA.hy926内皮细胞24小时。处理后,将细胞用胰蛋白酶消化,并在4℃和47×g下离心5分钟。沉淀物用于测定硫代巴比妥酸反应产物和一氧化氮水平。还评估了电子传递链活性和ATP水平。通过流式细胞术评估细胞内pH、细胞凋亡和线粒体膜去极化。通过HPLC-UV和总酚含量分析对MHS进行化学表征。通过2,2-二苯基-1-苦基肼自由基清除剂测定法评估MHS的抗氧化能力。为了确定MHS的体内急性毒性,使用0.2 mL不同浓度的MHS(10、50、100、250、500、750和1000 μg/mL)进行卤虫试验。通过在趋势曲线中插入50%(y = 50)死亡个体来获得LD值。
我们的数据表明,MHS能够避免高血糖诱导的氧化和亚硝化应激,显示出重要的抗氧化活性。然而,观察到MHS使四种电子传递复合物的活性降低了90%,从而降低了内皮细胞的ATP产生。在卤虫中进行的毒性试验中,MHS显示出轻度毒性(LD = 0.4 mg/mL)。在MHS中发现的主要化合物是表没食子儿茶素没食子酸酯、表儿茶素、芦丁、山奈酚和槲皮素。
这些数据提醒人们注意,含有高含量酚类化合物、能够避免氧化和亚硝化应激的补充剂可能具有双重作用,在具有抗氧化活性的同时,可能会对不同细胞类型产生毒性。