三脉菝葜醇提物及其不同极性萃取部位的抗肿瘤作用研究

目的:研究三脉菝葜醇提物及其不同极性萃取部位的抗肿瘤作用,为其抗肿瘤活性部位的筛选提供参考。方法:采用MTT法检测不同质量浓度的三脉菝葜醇提物及其石油醚、乙酸乙酯、正丁醇和水层萃取部位对人肝癌细胞株HepG2、人肺癌细胞株A549、人乳腺癌细胞株MCF-7和MDA-MB-231、人宫颈癌细胞株HeLa以及人卵巢癌细胞株HO-8910的增殖抑制率,并计算其半数抑制浓度(IC50)。将80只KM小鼠通过右前肢腋部皮下接种S180细胞悬液的方法复制荷瘤小鼠模型,并按体质量随机分为8组,即模型组(生理盐水,每天灌胃2次),环磷酰胺(阳性药物)组(0.025 g/kg,每天腹腔注射给药1次),三脉菝葜醇提物高、中、低剂量组和醇提物乙酸乙酯萃取部位高、中、低剂量组(以浸膏计给药剂量均分别为0.1、0.05、0.025 g/kg,每天灌胃2次),每组10只;各组小鼠均连续给药14 d;末次给药后禁食不禁水12 h,检测各组小鼠的抑瘤率、脾指数和胸腺指数。结果:体外试验表明,三脉菝葜及其不同极性萃取部位对6种肿瘤细胞的增殖抑制作用均呈现明显的量-效关系。其中以醇提物乙酸乙酯萃取部位的作用最强,其对各肿瘤细胞的IC50在40~210μg/mL范围内,对MCF-7细胞、MDA-MB-231细胞和HeLa细胞的IC50分别为70.56、83.58、44.67μg/mL。体内实验表明,各给药组小鼠的瘤质量较模型组均显著降低(P<0.05或P<0.01),并且三脉菝葜醇提物高剂量组和醇提物乙酸乙酯部位萃取部位高、中剂量组小鼠瘤质量显著低于环磷酰胺组(P<0.01);环磷酰胺组、三脉菝葜醇提物高剂量组和醇提物乙酸乙酯萃取部位高、中剂量组小鼠脾指数较模型组显著降低(P<0.01),环磷酰胺组小鼠胸腺指数较模型组和其余各给药组均显著升高(P<0.01)。结论:在4个极性部位中,三脉菝葜醇提物乙酸乙酯萃取部位的抗肿瘤效果最佳,并且免疫抑制作用较小。

Study on Anti-tumor Effect of Ethanol Extract of Smilax trinervula and Its Different Polar Extract Parts

OBJECTIVE:To study the anti-tumor effect of ethanol extract of Smilax trinervula and its different polar extract parts,and to provide reference for the screening of anti-tumor active parts. METHODS:MTT method was used to detect the inhibitory rates of different concentrations of ethanol extract of S. trinervula,its petroleum ether,ethyl acetate,n-butanol and water layer extract parts to the proliferation of human hepatocellular carcinoma cell line Hep G2,human lung cancer cell line A549,human breast cancer cell line MCF-7 and MDA-MB-231,human cervical cancer cell line He La and human ovarian cancer cell line HO-8910. Semi-inhibitory concentration(IC50)was calculated. A total of 80 KM mice were subcutaneously inoculated with S180 cell suspension in right forelimb axilla to induce tumor-bearing mice model. The mice were randomly divided into 8 groups:e.g.model group(normal saline,twice a day,i.g.),cyclophosphamide(positive drug) group(0.025 g/kg,once a day,i.p.),S.trinervula ethanol extract high-dose,medium-dose and low-dose groups,ethyl acetate part of ethanol extract high-dose,mediumdose and low-dose groups(0.1,0.05,0.025 g/kg by extract,twice a day,i.g.),with 10 rats in each group. The mice in each group were given medicine for consecutive 14 days. The inhibition rate,spleen index and thymus index of mice in each group were measured after fasting for 12 hours after the last administration. RESULTS:In vitro experiments showed that S. trinervula and different polar extracts inhibited the proliferation of 6 kinds of tumor cells in significant dose-effect manner,especially ethyl acetate part of ethanol extract. IC50 of it to tumor cells was 40-210 μg/m L,that of it to MCF-7,MDA-MB-231 and He La cells were 70.56,83.58,44.67 μg/m L,respectively. In vivo experiments showed that the tumor mass of mice were decreased significantly in each administrations(P<0.05 or P<0.01),the tumor mass of mice in S. trinervula ethanol extract highdose group,ethyl acetate part of ethanol extract high-dose and medium dose groups were significantly lower than cyclophosphamide group(P<0.01). The spleen index of mice was decreased significantly in cyclophosphamide group,S. trinervula ethanol extract high-dose group,ethyl acetate part of ethanol extract high-dose and medium dose groups,compared with model group(P<0.01);thymus index of mice in cyclophosphamide group was increased significantly,compared with model group and other administration groups(P<0.01). CONCLUSIONS:The ethyl acetate part of ethanol extract of S. trinervula has the best anti-tumor effect and less immunosuppressive effect.