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 脱皮卡皮罗纳（Capirona decorticans，茜草科(Rubiaceae)）在民间被广泛用于治疗疣、创伤、真菌病与疥疮，同时也是死藤水（Ayahuasca）茶的组分之一。尽管该物种的民间应用较为普遍，但针对其开展的植物化学与药理学研究仍十分有限。为此，本研究对C. decorticans叶片的乙醇提取物（ethanolic extract, EE）与氢甲醇馏分（hydromethanolic fraction, FM）中的酚类化合物进行了定量分析，二者的含量分别为406、293 mgEAG g⁻¹。通过LC-MS/MS-MMR-ESI技术鉴定出多种黄酮类成分，包括芹菜素（apigenin）、芦丁（rutin）、木犀草素（luteolin）、杨梅素（miricetin）、槲皮素（quercetin）、槲皮素-3-β-D-葡萄糖苷（quercetin-3-β-D-glucoside）以及槲皮苷（quercetrin）。此外，本研究以瑞士小鼠为实验对象，以环磷酰胺（cyclophosphamide, CP）作为DNA损伤与氧化应激诱导剂，通过体内实验评估了EE和FM的氧化应激水平及致突变/抗突变活性。实验过程中，小鼠连续15天每日给予EE或FM（250 mg kg⁻¹）预处理，随后腹腔注射CP（25 mg kg⁻¹）。分别检测肝、肾组织中的羰基化蛋白、抗坏血酸、过氧化氢酶与硫代巴比妥酸反应物质（thiobarbituric acid-reactive substances, TBARS）的含量。通过微核试验（Micronucleus Test）评估其致突变与抗突变活性。结果显示，FM可降低CP诱导的小鼠肝脏蛋白质羰基化水平，但对其余氧化应激标志物无显著影响。与单独使用CP的对照组相比，提取物处理组的CP诱导的微核多染红细胞频率均出现显著降低，其中EE组降低率达96%，FM组达71%。单独使用EE时则表现出致突变性。综上，C. decorticans叶片的EE和FM具备与其他物种相当的抗氧化潜力，未引发氧化应激或毒性，且具有保护与抗突变活性，尽管EE存在一定的致突变迹象。