福建绞股蓝皂苷的优化提取及其抗肝癌活性

Optimum Extraction and Anti-Hepatoma Activity of Gypenosides of Gynostemma pentaphyllum in Fujian Province

绞股蓝素有"南方人参"美誉,含有与人参皂苷相同骨架达玛烷型结构的三萜皂苷。通过不同体积分数乙醇溶液优化提取绞股蓝皂苷,测定其抗癌活性,为后续活性化合物的分离奠定基础。利用HPLC分析绞股蓝的皂苷成分并确定提取绞股蓝皂苷的乙醇体积分数,正交实验优化提取方法,进而MTT法测定提取的绞股蓝皂苷抗肝癌活性。HPLC分析确定用体积分数75%乙醇溶液提取绞股蓝总皂苷(GYP),体积分数45%乙醇提取极性绞股蓝皂苷(GYP1),体积分数90%乙醇提取弱极性绞股蓝皂苷(GYP2);正交实验结果表明在70℃水浴中以30质量的体积分数45%乙醇加热提取12 h为最佳条件提取GYP1,在60℃水浴中以30质量的体积分数90%乙醇加热提取6 h为最佳条件提取GYP2;利用MTT法测定GYP、GYP1和GYP2对肝癌细胞的抑制作用,GYP和GYP2对肝癌细胞SMCC7721具有抗癌活性,半抑制率IC50值分别为(165.83±14.12)μg/mL和(113.97±9.26)μg/mL,GYP1没有抗癌活性,表明绞股蓝抗癌药效成分主要是由弱极性绞股蓝皂苷发挥功效。

Gynostemma pentaphyllum is known as 'Southern Ginseng'.It contains triterpenoid saponins of the dammarane-type structure with the same skeleton of ginsenosides.In this manuscript,the saponin components of Gynostemma pentaphyllum were analyzed by HPLC,which choosed the concentration of ethanol extraction,and it was found that that adopted to the extract gypenosides(GYP) was from 75 % ethanol,polar gypenosides(GYP1) from 45% ethanol,and lower polar gypenosides(GYP 2) from 90% ethanol.Furthermore,the optimum extraction parameters for GYP 1 and GYP 2 were studied by Orthogonal method and listed as follows: GYP 1 was extracted with thirty fold-weight 45 % enthanol in 70 ℃ water bath for 12 h,and GYP 2 was extracted with thirty fold-weight 90% ethanol in 60 ℃ water bath for 6 h.Anti-hepatoma activity was determined tests by MTT and the results showed that GYP and GYP 2 can inhibit hepatoma cell SMCC7721 with IC 50 value 165.83 ±14.12 μg/mL and 113.97 ±9.26 μg/mL,respectively.However,GYP 1 without any anticancer activity.The result showed that Gynostemma pentaphyllum exhibited the anti-hepatoma activity caused by lower polar saponins.