藏药镰形棘豆醇提物酸碱分离及抗小鼠Lewis肺癌活性初步筛选

Separation of Alcohol Extracts from Tibetan Herb Oxytropis falcata by Acid and Alkali Treatment and Preliminary Screening of Tumor Suppresive Fraction on Mice Bearing Lewis Lung Cancer

目的筛选藏药镰形棘豆醇提物中对Lewis肺癌荷瘤小鼠具有抑瘤作用的活性部位。方法利用酸碱处理对镰形棘豆醇提物进行分离,并对所得碱性部位(AlF)和酸性部位(AcF)进行定性、定量分析。建立Lewis肺癌C57BL/6小鼠模型,随机分为模型组,环磷酰胺(CTX)组(20 mg·kg^(-1)),AlF高、低剂量组(180,90 mg·kg^(-1)),AcF高、低剂量组(180,90 mg·kg^(-1)),连续给药10 d;通过测定小鼠瘤体质量、抑瘤率、胸腺指数、脾指数来比较AlF、AcF的抑瘤作用;HE染色,显微镜下观察各组肿瘤细胞的形态学变化。结果 AcF主要含黄酮类成分,含量为18.9%;AlF主要为生物碱成分,含量为64.1%。AcF高剂量组降低雌性、雄性小鼠瘤体质量的作用明显,与模型组比较,差异有统计学意义(P<0.01,P<0.05),抑瘤率分别为36.3%、32.8%;AlF高剂量组降低雌性小鼠瘤体质量的作用明显,与模型组比较,差异有统计学意义(P<0.05),抑瘤率为29.7%。与模型组比较,CTX组小鼠的脾脏、胸腺指数均明显降低,差异有统计学意义(P<0.05,P<0.01);而AcF、AlF各剂量组与模型组比较,脾脏、胸腺指数差异均无统计学意义(P>0.05)。肿瘤组织病理形态学观察显示AcF、AlF各给药组肿瘤细胞出现不同程度的坏死。结论镰形棘豆醇提物AlF、AcF均对Lewis肺癌荷瘤小鼠具有明显的抗肿瘤作用,黄酮类成分的作用比生物碱略强,其抗肿瘤机制还有待进一步研究。

Objective To screen the tumor suppressive fraction from ethanol extracts of Tibetan herb Oxytropis falcata on mice bearing Lewis lung cancer. Methods O. falcata was extracted with ethanol and separated into acid alcohol extract （AcF） and alkaline alcohol extract （A1F）. These two fractions were analyzed qualitatively and quantitatively. The Lewis lung cancer bearing mice were prepared and randomly divided into model group, CTX group （20 mg·kg^-l）, high and low dosages of AIF （180, 90 mg·kg^-1） groups, high and low dosages of AcF （180, 90 mg·kg-1） groups, followed by a successive administration for 10 days. Tumor weight, tumor inhibition, thymus index and spleen index of the mice were measured to compare the antitumor effect of AcF and A1F. Cells were stained with HE and the morphological status of cells were observed under microscope. Results The content of flavonoids in AcF was 18.9 %, and the alkaloids in A1F was 64.1%. Tumor inhibition of high dose group of O. falcata AcF was 36.3 % on female mice, while which was 32.8 % on male mice. Compared with model group, the difference was statistically significant （P 〈 0.05, P 〈 0.01 ）. Tumor inhibition of high dose group of O. falcata A1F was 29.7 % on female mice, while was 25.6 % on male mice, showed significant differences compared with model group （P 〈 0.05）. Compared with model group, immune indexes （spleen index and thymus index） of positive group reduced markedly （P 〈 0.05, P 〈 0.01 ）. No statistically significant differences were found in immune indexes of model group and those of O. falcata AcF and A1F. Morphological observation of tumor tissues showed that different degrees of necrosis appeared in AcF and A1F groups. Conclusion The results indicate that both AcF and A1F fractions of O. fa!cata have obvious antitumor effect in Lewis lung cancer bearing mice. Tumor suppressive effect of AcF is slightly stronger than that of A1F. Further study is needed to explore the antitumor mechanism.