Anticrotalic and antitumoral activities of gel filtration fractions of aqueous extract from Tabernaemontana catharinensis (Apocynaceae)
Anticrotalic and antitumoral activities of gel filtration fractions of aqueous extract from Tabernaemontana catharinensis (Apocynaceae)
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DOI：
10.1016/j.cca.2003.10.012
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发表时间：
2004-01-01
影响因子：
3.9
通讯作者：
Sampaio, SV
中科院分区：
文献类型：
--
作者：
de Almeida, L;Cintra, ACO;Sampaio, SV
巴豆腐蛇毒致死率高的主要原因是其毒素作用于神经肌肉接头，抑制了乙酰胆碱释放的调节机制，从而导致动物运动和呼吸瘫痪，进而导致动物死亡。我们最近证实，华山牛蛙的水提物(AE)能抑制可怕圆线虫毒液的致死活性。提取液经Sephadex G-10凝胶过滤，得到8个组分，P-I至P-VIII，并测定其细胞毒性和致死性。组分P-VII[2 mg/100g大鼠/50毫升生理盐水(Ss)]肌肉注射(肌肉注射)20 S后，毒液(240杯)或巴豆毒素(200杯/50毫升)均中和两者的2LD50的致死活性。用Mosmann比色法测定了组分P-I、P-VI和P-VIII(5.0 mg/100g大鼠/50MUSS)孵育24 h后对人乳腺癌SK-BR-3细胞的体外抗肿瘤活性。组分P-VII的主要成分为12-甲氧基-4-甲基胡萝卜素。这些结果表明，青蒿素的抗蛇毒活性和抗肿瘤活性分别是由组分P-VII和组分P-I、P-VI和P-中的不同物质发挥的，薄层层析法分析它们的染色和Rf特征是不同的。结果还表明，Sephadex G-10凝胶过滤的初步分级是从青藤中分离生物活性物质的一个很好的选择。(C)2003 Elsevier Inc.保留所有权利。
The high mortality caused by Crotalus durissus terrificus snake venom is mainly due to crotoxin, which acts on the neuromuscular junction inhibiting the mechanism mediating acetylcholine release, thus leading to motor and respiratory paralysis and subsequently to animal death. We recently demonstrated that the aqueous extract (AE) of Tabernaemontana cathatinensis can inhibit the lethal activity of C. d. terrificus venom. Eight fractions, P-I to P-VIII, were obtained by gel filtration of the extract on Sephadex G-10, and assayed for lethality and cytotoxicity. Fraction P-VII [2.0 mg/100 g rat/50 mul saline solution (ss)] injected intramuscularly (i.m.) 20 s after the venom (240 mug) or crotoxin (200 mug/50 mul ss) neutralized the lethal activity of 2 LD50 of both. Fractions P-I, P-VI and P-VIII (5.0 mg/100 g rat/50 mul ss) presented potent antitumoral activity in vitro against cells from human breast carcinoma (SK-BR-3) after 24 h incubation, as measured by Mosmann colorimetric method. Fraction P-VII contains 12-methoxy-4-methylvoachalotine as its major component. These results demonstrate that the antivenom and antitumoral activities of the AE of T catharinensis are exerted by different substances present in fraction P-VII and fractions P-I, P-VI, and P-VIII, respectively, whose characteristics are distinct in terms of staining and Rf when analyzed by thin layer chromatography. The results also show that a preliminary fractionation by Sephadex G-10 gel filtration is a good option as a first step for isolation of biologically active substances from T catharinensis. (C) 2003 Elsevier Inc. All rights reserved.