吴兴铁线莲总黄酮的提取及体外抗肿瘤活性研究

Study on Flavonoids Extraction from Clematis Huchouensis Tamura and Its in Vitro Antineoplasmic Activity

关键词:  吴兴铁线莲  黄酮  抗肿瘤活性  增殖抑制率

Key words:  Clematis huchouensis Tamura  flavone  antineoplasmic activity  proliferation inhibition rate

摘要:

目的 对吴兴铁线莲总黄酮成分进行分离提取，研究其抗肿瘤活性。方法 采用紫外可见分光光度法测定总黄酮含量，以聚酰胺层析柱层析乙醇洗脱提纯吴兴铁线莲中的黄酮成分得到黄酮浸膏，通过MTT比色法用酶标仪测定光吸收值，计算半数抑制浓度(IC₅₀)，观察100~25 600 mg·L^-1不同浓度的铁线莲黄酮溶液对小鼠骨髓瘤细胞SP2/0体外增殖的作用，来反映其抑制肿瘤细胞生长的灵敏性。结果 铁线莲浸膏稀释液经芦丁标准曲线吸光度测定，浸膏中黄酮含量为82.7%，铁线莲干粉中黄酮含量为47 mg·g^-1，2.843 g铁线莲浸膏经洗脱提纯得到黄酮浸膏0.158 g。提纯后铁线莲黄酮溶液对骨髓瘤细胞SP2/0的增殖抑制率随总黄酮浓度的升高而增加，最后趋于稳定，其IC₅₀值为408.8 mg·L^-1。结论 吴兴铁线莲黄酮提取物能抑制骨髓瘤细胞SP2/0体外增殖，具有特异性强、灵敏度高的抗肿瘤活性，其良好的量效关系可为防治肿瘤潜在价值的开发应用提供依据。

Abstract:

OBJECTIVE To study the antineoplasmic activity of the flavonoids component extracted from Clematis huchouensis Tamura. METHODS The content of total flavonoids was determined by Ultraviolet-Visible Spectrophotometry. Polyamide column chromatography combined with ethanol elution was used to purify the flavonoids of Clematis huchouensis Tamura to obtain its extract. ELIASA was applied in measuring optical density value with the adoption of MTT colorimetric method. The meadian inhibitory concentration(IC₅₀) was determined. The effect of Clematis huchouensis Tamura flavonoids solution with the concentration varying between 100~25 600 mg·L^-1 impacting on in vitro proliferation of myeloma cells SP2/0 in mice was observed. So that its sensibility in inhibiting tumor cell growth was realized. RESULTS The flavonoids in the Clematis huchouensis Tamura extract was 82.7% by absorbance measurement of rutin standard curve in its dilution. The flavonoid content in Clematis huchouensis Tamura dry powder was 47 mg·g^-1. Flavonoid extract of 0.158 g was obtained by elution and purification of 2.843 g Clematis huchouensis Tamura extract. Inhibition rate of purified Clematis huchouensis Tamura flavonoids solution impact on proliferation of myeloma cell SP2/0 increased along with the upward tendency of total falvonoids and eventually tend to be stable. The IC₅₀ value was 408.8 mg·L^-1. CONCLUSION Flavonoids extraction from Clematis huchouensis Tamura demonstrate antineoplasmic activity with strong specificity and high sensitivity through inhibition of myeloma cells SP2/0 proliferation in mice. Its favourable dose-effect provides reference for further application of potential role in the prevention and treatment of tumor.