First phytochemical and biological study of the ethanolic extract from leaves of Capirona decorticans (Rubiaceae)

ABSTRACT Capirona decorticans (Rubiaceae) is popularly used to treat warts, wounds, mycoses and scabies, and is also a component of the Ayahuasca tea. Despite its popular use, the phytochemical and pharmacological research on this species is limited. Therefore, this work quantified phenolic compounds in the ethanolic extract (EE) and hydromethanolic fraction (FM) (406, 293 mgEAG g-1, respectively) from leaves of C. decorticans. We identified flavonoids by LC-MS/MS-MMR-ESI (apigenin, rutin, luteolin, miricetin, quercetin, quercetin-3-β-D-glucoside, quercetrin), and evaluated oxidative stress and mutagenic/antimutagenic effect of EE and FM through an in vivo experiment using Swiss mice and cyclophosphamide (CP) as an inducer of DNA damage and oxidative stress. Mice were pretreated for 15 consecutive days with EE or FM (250 mg kg-1) and then intraperitoneally injected with CP (25 mg kg-1). Carbonylated proteins, ascorbic acid, catalase and thiobarbituric acid-reactive substances were measured in hepatic and renal tissues. The mutagenic/antimutagenic effect was evaluated through the Micronucleus Test. Protein carbonylation in the liver of animals exposed to CP was reduced by FM. There was no significant effect on other markers of oxidative stress. The groups treated with the extracts showed a significant percentage reduction (EE = 96% and FM = 71%) in the frequency of micronucleated polychromatic erythrocytes induced by CP. EE showed mutagenicity when used alone. The EE and FM of C. decorticans leaves showed antioxidant potential equivalent to that observed in other species, did not cause oxidative stress, nor toxicity, and had a protective and antimutagenic effect, although the EE showed signs of mutagenicity.

摘要：卷边帽蕊木（Capirona decorticans，茜草科Rubiaceae）在民间常用于治疗疣、伤口、真菌病及疥疮，同时也是死藤水（Ayahuasca）茶的组分之一。尽管其应用广泛，但目前针对该物种的植物化学与药理学研究仍较为有限。为此，本研究对卷边帽蕊木叶片的乙醇提取物（ethanolic extract, EE）与水甲醇馏分（hydromethanolic fraction, FM）中的酚类物质进行了定量分析，其含量分别为406、293 mgEAG·g⁻¹。本研究通过液相色谱-串联质谱-多反应监测-电喷雾电离（LC-MS/MS-MMR-ESI）技术鉴定出多种黄酮类化合物，包括芹菜素、芦丁、木犀草素、杨梅素、槲皮素、槲皮素-3-β-D-葡萄糖苷及槲皮苷；并以环磷酰胺（cyclophosphamide, CP）作为DNA损伤与氧化应激诱导剂，通过瑞士小鼠体内实验评估了EE与FM的氧化应激及致突变/抗突变活性。实验小鼠先连续15天接受EE或FM处理（剂量为250 mg·kg⁻¹），随后腹腔注射CP（剂量为25 mg·kg⁻¹）。研究人员对小鼠肝、肾组织中的羰基化蛋白、抗坏血酸、过氧化氢酶及硫代巴比妥酸反应物质进行了检测，并通过微核试验评估样本的致突变/抗突变活性。结果显示，FM可降低CP暴露小鼠肝脏中的蛋白质羰基化水平，其余氧化应激标志物未出现显著变化。提取物处理组小鼠的CP诱导的微核多染红细胞发生率均显著降低，其中EE组降低率为96%，FM组为71%。单独使用EE时，其表现出致突变性。卷边帽蕊木叶片的EE与FM均具有与其他物种提取物相当的抗氧化活性，未引发氧化应激或毒性，且具备保护与抗突变作用，不过EE单独使用时存在致突变迹象。