Departamento de Bioquímica, Instituto de Biologia Roberto Alcantara Gomes, Centro Biomédico, Universidade do Estado do Rio de Janeiro, Av. Professor Manoel de Abreu, 444, PAPC, 4o andar, CEP 20550-170, Rio de Janeiro, RJ, Brazil.
J Ethnopharmacol. 2011 Nov 18;138(2):513-22. doi: 10.1016/j.jep.2011.09.051. Epub 2011 Oct 12.
Baccharis trimera (Less) DC. (Asteraceae), popularly known in Brazil as "carqueja", have been used in folk medicine to treat gastrointestinal, hepatic and renal diseases, and inflammatory processes as rheumatism.
To evaluate the in vitro and in vivo toxicological effects of anti-inflammatory Baccharis trimera aqueous extract and fractions.
Aqueous extract of Baccharis trimera (AEBt) was produced by infusion in boiling water. After lyophylization AEBt was extracted with 80% ethanol, originating the ethanolic supernatant fraction (EFBt) and the aqueous sediment fraction (AFBt). Anti-inflammatory properties of AEBt, EFBt or AFBt (3, 30 or 300 μg/kg b.w.) were evaluated by the carrageenan-induced mouse paw edema using indomethacin (10mg/kg) as positive control. The growth of rat hepatoma cells (HTC) and human embryo kidney epithelial cells (HEK) was determined by protein staining assay. Cytotoxicity was assayed by the tetrazolium salt (MTT) reduction. Cyclosporin was used as reference cytotoxic drug for spleen cells and doxorubicin for HTC and HEK cells. For in vivo toxicological evaluation SW male mice were daily and oral (gavage) treated with extract/fractions at 4.2mg/kg or 42 mg/kg during 15 days. After treatment liver or kidney cells were submitted to comet assay to determine the DNA damage index, and the glutathione S-transferase activity was assayed towards ETHA (class Pi) and CDNB (several classes). Mutagenicity was evaluated by the Ames test using Salmonella typhimurium strains TA97, TA98, TA100, and TA102.
The anti-inflammatory effects of EFBt were higher than those of AEBt or AFBt. Mice treatment (3-300 μg/kg) with AFBt reduced the paw edema (3h) at lower levels (29.2-37.3%; P<0.01), than those observed for AEBt (44.7-54.2%; P<0.001), EFBt (49.3-58.2%; P<0.001) or indomethacin (64.6%, P<0.001, 10mg/kg). The growth of kidney cells (HEK) was inhibited by AEBt (IC(50) 182.6 μg/ml), EFBt (IC(50) 78.1 μg/ml) and AFBt (IC(50) 86.2 μg/ml), with lower effects on HTC hepatic cell (IC(50) 308.8 μg/ml, 396.5 μg/ml and 167.9 μg/ml, respectively). As evaluated by MTT test, AFBt exhibited cytotoxicity for HEK cells (IC(50) 372.5 μg/ml), but none for HTC ones; by the way, AFBt stimulated spleen cells (EC(50) 2.2 μg/ml) while cyclosporine, a cytotoxic reference drug inhibited them with IC(50) of 0.42 μg/ml; the IC(50) for doxorubicin for HEK and HTC cells was 0.28 μg/ml and 14.4 μg/ml, respectively, at 96h. No mutagenic potential was observed. Mice treatment with AEBt or AFBt at 42 mg/kg for 15 days altered the kidney relative weight, but not at 4.2mg/kg. Baccharis trimera did not change liver, spleen or popliteal lymph node relative weight. DNA damage index of kidney cells was observed on mice treated with AEBt/AFBt, but not on animals treated with EFBt, while DNA lesions were detected on liver cells only after AFBt treatment. The general activities of hepatic GST and Pi GST were reduced by EFBt and AFBt treatment, respectively.
Baccharis trimera did not show mutagenicity, inhibited the GST activity, a hepatic detoxification enzyme, and induced in vivo (genotoxicity) and in vitro toxicological effects to kidney cells.
Baccharis trimera（Less）DC.（菊科），在巴西俗称“carqueja”，已被民间医学用于治疗胃肠道、肝和肾疾病以及风湿性炎症等疾病。
评估抗炎 Baccharis trimera 水提取物和馏分的体外和体内毒理学效应。
通过在沸水中浸泡产生 Baccharis trimera 水提取物（AEBt）。在 lyophylization 后，AEBt 用 80%乙醇提取，产生乙醇上清液部分（EFBt）和水沉淀部分（AFBt）。通过角叉菜胶诱导的小鼠爪肿胀评价 AEBt、EFBt 或 AFBt（3、30 或 300μg/kg b.w.）的抗炎特性，使用吲哚美辛（10mg/kg）作为阳性对照。通过蛋白质染色测定法测定大鼠肝癌细胞（HTC）和人胚肾上皮细胞（HEK）的生长。通过四唑盐（MTT）还原测定细胞毒性。环孢素用于脾细胞的参考细胞毒性药物，阿霉素用于 HTC 和 HEK 细胞。为了进行体内毒理学评价，SW 雄性小鼠每天口服（灌胃）以 4.2mg/kg 或 42mg/kg 的剂量用提取物/馏分处理 15 天。处理后，将肝或肾细胞进行彗星试验以确定 DNA 损伤指数，并测定 GST 对 ETHA（Pi 类）和 CDNB（多种类别）的活性。使用鼠伤寒沙门氏菌 TA97、TA98、TA100 和 TA102 菌株通过艾姆斯试验评估致突变性。
EFBt 的抗炎作用高于 AEBt 或 AFBt。用 AFBt（3-300μg/kg）处理小鼠可降低 3h 时的爪肿胀（37.3%；P<0.01），低于 AEBt（44.7-54.2%；P<0.001）、EFBt（49.3-58.2%；P<0.001）或吲哚美辛（64.6%；P<0.001，10mg/kg）的水平。AEBt（IC50182.6μg/ml）、EFBt（IC5078.1μg/ml）和 AFBt（IC5086.2μg/ml）抑制肾细胞（HEK）的生长，而对肝细胞（HTC）的影响较小（IC50308.8μg/ml、396.5μg/ml 和 167.9μg/ml）。通过 MTT 试验评估，AEBt 对 HEK 细胞表现出细胞毒性（IC50372.5μg/ml），而对 HTC 细胞没有影响; 另一方面，AEBt 刺激脾细胞（EC502.2μg/ml），而环孢素作为细胞毒性参考药物抑制它们的 IC50 为 0.42μg/ml; 阿霉素对 HEK 和 HTC 细胞的 IC50 分别为 0.28μg/ml 和 14.4μg/ml，作用 96h。未观察到致突变性。用 42mg/kg 的 AEBt 或 AFBt 处理小鼠 15 天改变了肾脏的相对重量，但在 4.2mg/kg 时没有改变。Baccharis trimera 没有改变肝脏、脾脏或腘淋巴结的相对重量。用 AEBt/AFBt 处理的小鼠观察到肾细胞的 DNA 损伤指数，但用 EFBt 处理的动物没有观察到，而仅在用 AFBt 处理后才检测到肝细胞的 DNA 损伤。EFBt 和 AFBt 处理分别降低了肝 GST 和 Pi GST 的一般活性。
Baccharis trimera 没有表现出致突变性，抑制了 GST 活性，一种肝解毒酶，并在体内（遗传毒性）和体外肾细胞毒性作用。
