Oliveira Mariana Canevari, Lemos Larissa Maria Scalon, de Oliveira Ruberlei Godinho, Dall'Oglio Evandro Luiz, de Sousa Júnior Paulo Teixeira, de Oliveira Martins Domingos Tabajara
Department of Basic Sciences in Health, Faculty of Medicine, Federal University of Mato Grosso (UFMT), Av. Fernando Correa da Costa, no. 2367, Boa Esperança, Cuiabá, Mato Grosso 78060-900, Brazil.
Natural Products Chemistry Research Laboratory, Department of Chemistry, Institute of Exact and Earth Sciences, Federal University of Mato Grosso (UFMT), Av. Fernando Correa da Costa, no. 2367, Boa Esperança, Cuiabá, Mato Grosso, 78060-900, Brazil.
J Ethnopharmacol. 2014 Aug 8;155(1):30-8. doi: 10.1016/j.jep.2014.06.019. Epub 2014 Jun 13.
Calophyllum brasiliense Camb., Clusiaceae, is commonly known as "guanandi" and its stem bark is used in Brazilian traditional medicine to treat rheumatism, vein problems, hemorrhoids and gastric ulcers. The aim of this study was to evaluate the toxicity of hexane extract of Calophyllum brasiliense stem bark (HECb) using in vitro and in vivo experimental models.
In vitro toxicity was evaluated by Alamar Blue cytotoxicity assay and micronucleus test, using Chinese hamster ovary (CHO-k1) epithelial cells. in vivo toxicity was evaluated by oral acute and subchronic toxicity assays. In the oral acute toxicity screening, a single dose of HECb was administered to mice at doses ranging from 250 to 1000 mg/kg. In the subchronic study, HECb was administered orally for 30 days to Wistar rats at doses of 100 mg/kg and 500 mg/kg. Phytochemical analyses were performed by HPLC/UV-vis, secondary metabolites were quantified by spectrophotometric methods.
HECb presented IC50=119.94±4.31 µg/mL after a 24 h cytotoxicity test using CHO-k1 cells, showing low cytotoxicity. However, when the cells were exposed to HECb for 72 h, the IC50 value was 8.39±2.00 µg/mL, showing in this case, a pronounced cytotoxic effect. In the oral acute toxicity studies, doses up to 500 mg/kg of HECb did not cause any changes in both male and female mice. At 1000 mg/kg, male mice showed signs typical of depression and stimulation that were reversed at 72 h. Besides, female mice were more sensitive to the toxic effect of HECb at 1000 mg/kg, which initially presented typical agitation signals, followed by depression signals, leading to death of all the animals at 24h. In subchronic assay with rats, HECb administered orally at doses of 100 and 500 mg/kg did not cause significant changes in all clinical parameters evaluated. Histopathological analyses showed no deleterious effect in the vital organs of rats. Preliminary phytochemical analysis revealed the presence of phenolic compounds, steroids, and volatile coumarins. Analysis by HPLC showed two major peaks characteristic of chromanones.
In vitro toxicological tests showed that HECb exhibited cytotoxicity especially after 72 h of exposition, and mutagenicity on the highest tested dose. The in vivo studies demonstrated that HECb produced some toxicity signs at the highest dose tested, particularly, in the acute toxicity test but showed no significant signs of toxicity in the subchronic assay. Based on these and previous pharmacological studies, it is possible to say that HECb did not exhibit significant toxicity at its effective dose. This suggests that HECb is relatively safe in humans at its effective dose.
巴西红厚壳（Calophyllum brasiliense Camb.），藤黄科植物，俗称“瓜南迪”，其茎皮在巴西传统医学中用于治疗风湿病、静脉问题、痔疮和胃溃疡。本研究的目的是使用体外和体内实验模型评估巴西红厚壳茎皮己烷提取物（HECb）的毒性。
使用中国仓鼠卵巢（CHO-k1）上皮细胞，通过alamar蓝细胞毒性试验和微核试验评估体外毒性。通过口服急性和亚慢性毒性试验评估体内毒性。在口服急性毒性筛选中，以250至1000mg/kg的剂量向小鼠单次给药HECb。在亚慢性研究中，以100mg/kg和500mg/kg的剂量向Wistar大鼠口服给药HECb 30天。通过HPLC/UV-vis进行植物化学分析，通过分光光度法对次生代谢物进行定量。
使用CHO-k1细胞进行24小时细胞毒性试验后，HECb的IC50 = 119.94±4.31μg/mL，显示出低细胞毒性。然而，当细胞暴露于HECb 72小时时，IC50值为8.39±2.00μg/mL，在这种情况下显示出明显的细胞毒性作用。在口服急性毒性研究中，高达500mg/kg的HECb剂量对雄性和雌性小鼠均未引起任何变化。在1000mg/kg时，雄性小鼠表现出典型的抑郁和兴奋迹象，在72小时时恢复正常。此外，雌性小鼠对1000mg/kg的HECb毒性作用更敏感，最初表现出典型的激动信号，随后是抑郁信号，导致所有动物在24小时死亡。在大鼠亚慢性试验中，以100mg/kg和500mg/kg的剂量口服给药HECb对所有评估的临床参数均未引起显著变化。组织病理学分析显示对大鼠重要器官无有害影响。初步植物化学分析显示存在酚类化合物、类固醇和挥发性香豆素。HPLC分析显示两个主要的色原酮特征峰。
体外毒理学试验表明，HECb尤其在暴露72小时后表现出细胞毒性，并且在最高测试剂量下具有致突变性。体内研究表明，HECb在最高测试剂量下产生了一些毒性迹象，特别是在急性毒性试验中，但在亚慢性试验中未显示出明显的毒性迹象。基于这些和先前的药理学研究，可以说HECb在其有效剂量下未表现出显著毒性。这表明HECb在其有效剂量下对人类相对安全。
