Mesia G K, Tona G L, Nanga T H, Cimanga R K, Apers S, Cos P, Maes L, Pieters L, Vlietinck A J
University of Kinshasa, Faculty of Pharmaceutical Sciences, PO BOX 212, Kinshasa XI, People's Republic of Congo.
J Ethnopharmacol. 2008 Feb 12;115(3):409-15. doi: 10.1016/j.jep.2007.10.028. Epub 2007 Dec 18.
To evaluate in vitro the antiprotozoal and cytotoxic activities of 80% methanol extract from 45 medicinal plants collected in Sankuru (Democratic Republic of Congo) against Trypanosoma brucei brucei, Trypanosoma cruzi and the chloroquine-sensitive Ghanaian strain of Plasmodium falciparum, and MRC-5 cell lines respectively.
Different extracts were obtained by maceration of each plant part used with 80% methanol for 24h. The mixture was filtered and evaporated in vacuo to give corresponding dried extract. The activity against Trypanosoma brucei brucei and Trypanosoma cruzi were performed in 96 well tissue plates each containing 10 microl aqueous plant extract dilutions (100 to 0.01 microg/ml) with 10 microl of the parasite suspension cultured in Hirumi medium supplemented with 10% foetal calf serum, a solution of 2% penicillin/streptomycin (2% P/S) After 4 days incubation with Almar blueâ solution, fluorescence was measured at 500 nm emission and 530 nm excitation and results expressed as percentage reduction in parasite compared to control wells. The antiplasmodial activity of was assessed in vitro against the chloroquine-sensitive Ghanaian strain of Plasmodium falciparum cultured in RPMI-1640 medium by the lactate deshydrogenase assay in the presence of plant extracts (50 to 0.01 microg/ml). Cell-lines MRC-5 were cultured in MEM medium supplemented with 20mM l-glutamine, 16.5mM NaHCO(3), 5% foetal calf serum and 2% P/S solution. After 4h incubation, cell proliferation/viability was spectrophotomecally assessed at 540 nm after addition of MTT. In each assay, the IC50 value for each sample was derived by the drug concentration-response curves.
The extracts from Alcornea cordifolia leaves, Momordica charantia whole plant, Omphalocarpum glomerata, root bark and Piptadia africanum stem bark showed good antiprotozoal activity against Trypanosoma brucei brucei with IC50 values from 0.7 to 7 microg/ml. Only Piptadenia africanum extract showed a pronounced antiprotozoal activity against Trypanosoma cruzi (IC50=4.0+/-06 microg/ml). The extracts from Alchornea cordifolia, Polyathia swaveleons stem bark, Sapium cornutum stem bark and Triclisia giletii stem bark exhibited a pronounced antiplasmodial activity against P. falciparum Ghanaian strain with IC50 values ranging from 0.5 to 3.0 microg/ml. Piptadenia africanum extract was the most cytotoxic sample (CC50=0.25 microg/ml) with poor selectivity against all selected protozoa (SI<10) while other active extracts did not show a significant cytotoxic effect against MCR-5 cell-lines with good selectivity according to the case.
These active plant extracts are selected for extensive studies leading to the isolation of active constituents.
在体外评估从刚果民主共和国桑库鲁收集的45种药用植物的80%甲醇提取物对布氏布氏锥虫、克氏锥虫以及氯喹敏感的加纳恶性疟原虫株和MRC - 5细胞系的抗寄生虫和细胞毒性活性。
将每种植物部分与80%甲醇浸渍24小时获得不同提取物。混合物过滤并真空蒸发得到相应的干燥提取物。对布氏布氏锥虫和克氏锥虫的活性在96孔组织培养板中进行,每孔含有10微升植物提取物水溶液稀释液(100至0.01微克/毫升)以及10微升在补充有10%胎牛血清、2%青霉素/链霉素溶液(2% P/S)的希鲁米培养基中培养的寄生虫悬浮液。与阿尔玛蓝溶液孵育4天后,在发射波长500纳米和激发波长530纳米处测量荧光,结果以与对照孔相比寄生虫减少的百分比表示。通过在植物提取物(50至0.01微克/毫升)存在下的乳酸脱氢酶测定法,在体外评估对在RPMI - 1640培养基中培养的氯喹敏感的加纳恶性疟原虫株的抗疟活性。MRC - 5细胞系在补充有20毫摩尔L - 谷氨酰胺、16.5毫摩尔碳酸氢钠、5%胎牛血清和2% P/S溶液的MEM培养基中培养。孵育4小时后,加入MTT后在540纳米处通过分光光度法评估细胞增殖/活力。在每个实验中,每个样品的IC50值通过药物浓度 - 反应曲线得出。
心叶阿尔柯纳叶提取物、苦瓜全株提取物、球果油瓜、根皮提取物以及非洲匹塔迪亚茎皮提取物对布氏布氏锥虫显示出良好的抗寄生虫活性,IC50值为0.7至7微克/毫升。只有非洲匹塔迪亚提取物对克氏锥虫显示出显著的抗寄生虫活性(IC50 = 4.0±0.6微克/毫升)。心叶阿尔柯纳、多花暗罗茎皮、角果乌桕茎皮和吉氏曲蕊马钱茎皮提取物对加纳恶性疟原虫株表现出显著的抗疟活性,IC50值范围为0.5至3.0微克/毫升。非洲匹塔迪亚提取物是细胞毒性最大的样品(CC50 = 0.25微克/毫升),对所有选定的原生动物选择性较差(SI < 10),而其他活性提取物根据情况对MCR - 5细胞系未显示出显著的细胞毒性作用且选择性良好。
选择这些活性植物提取物进行深入研究以分离活性成分。