绞股蓝的化学成分研究
|
摘要
绞股蓝为葫芦科(Cucurbitaceae)绞股蓝属植物kg绞股蓝[Gynostemma pentaphyllum (Thunb.) Makino]的干燥全草,为多年生攀缘草本,主要分布于中国、日本、朝鲜等国。绞股蓝是五加科外具有人参皂苷资源的植物之一,具有滋补保健、抗癌防衰、增强体质和改善脂质代谢等多种功能,被誉为“南方人参”。
本文在综述了绞股蓝的化学成分和药理活性研究的基础上,对其进行了系统的化学成分研究及抗肿瘤活性研究。采用反复硅胶柱色谱法、大孔树脂、Sephadex LH-20、反相ODS、半制备型HPLC等多种色谱手段从绞股蓝的75%乙醇提取物中,分离得到38个化合物。并通过理化性质、波谱数据分析(IR、NMR、MS等)和化学方法鉴定了它们的结构,分别为：(23S)-21R-O-正丁基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-比喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(1),(23S)-21S-O-正丁基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(2),(23S)-21R-O-正丁基-19-醛基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(3),(23S)-21S-O-正丁基-19-醛基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(4),(23S)-21R-O-正丁基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(5),(20S)-3β,20,21-三羟基达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(6),长梗绞股蓝皂苷Ⅰ(7),α-菠菜甾醇(8),β-谷甾醇(9),α-菠菜甾醇3-O-β-D-吡喃葡萄糖苷(10),胡萝卜苷(11),绞股蓝皂苷Ⅲ(人参皂苷Rb1,12),绞股蓝皂苷Ⅳ(人参皂苷Rb3,13),绞股蓝皂苷Ⅷ(人参皂苷Rd,14),绞股蓝皂苷ⅩⅣ(15),绞股蓝皂苷Ⅻ(人参皂苷F2,16),绞股蓝皂苷ⅩⅢ(17),芸香苷(18),丙二酸(19),(20R,23R)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-6-O-乙基-β-D-吡喃葡萄糖苷(20),(20S,23S)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-6-O-乙基-β-D-吡喃葡萄糖苷(21),绞股蓝皂苷XLIX (22),(20S)-3β,20,21-三羟基达玛烷-24-烯3-O-{[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖基}-21-O-β-D-吡喃葡萄糖苷(23),(23S,23S)-19-醛基-3β,20ξ,21,26-四羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(24),(23R,23R)-19-醛基-3β,20ξ,21,26-四羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(25),(20R,23R)-19-醛基-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(26),(20S,23S)-19-醛基-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(27),(20S)-3β,20,21-三羟基达玛-23,25-二烯3-O-{[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖基}-21-O-β-D-吡喃葡萄糖苷(28),(20S)-19-醛基-3β,20,21-三羟基达玛-23,25-二烯3-O-{[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖基}-21-O-β-D-吡喃葡萄糖苷(29),(23S)-21R-O-乙基-19-醛基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(30),(23S)-19-醛基-3β,20ξ,21ξ-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(31),(20S)-19-醛基-3β,20,21ξ,25-四羟基-21,24ξ-环达玛烷3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(32),(20R)-19-醛基-3β,20,21ξ,23ξ-四羟基-21,24ξ-环达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(33),(20R,23R)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(34),(20S,23S)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(35),(20S,23S)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(36), (20R,23R)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[β-D-吡喃木糖基(1→3)][β-D-吡喃葡萄糖苷(37),槲皮素(38)。其中未见文献报道的新化合物14个,分别为化合物1-6、24-29、36和37。
采用MTT法,对从绞股蓝中得到的部分化合物的体外肿瘤抑制活性进行了初步筛选和评价。化合物1-5,26,33,36-37对细胞株HL-60均具有不同程度的细胞生长抑制作用,其中化合物3活性最强,IC50值为8.45μmol/L.化合物1-5,7对细胞株Colon205和Du145不同浓度下均具有生长抑制作用,其中化合物2对Colon 205的活性最强,IC50值为22.95μmol/L,化合物3对Du145的活性最强,IC50值为17.41μmol/L。化合物20-21,34-35对细胞株A2780和OVCAR-3在不同浓度下均具有生长抑制作用,其中化合物20和34对A2780有较强的活性,其IC50均为10.4μmol/L,化合物20对OVCAR-3有较强的活性,其IC50为2.8μmol/L。
Gynostemma pentaphyllum (Thunb.) Makino (Cucurbitaceae) is a herbal medicine with anticancer activity, widely distributed in China, Korea and Japan. It is famous as "Southen Ginseng". The biological active constituents are dammarane-type glycosides, called gypenosides, which are structurally correlated to the ginseng saponins. Pharmacological investigation demonstrated that extracts of this plant exhibited a variety of biological effects, such as anticancer, anti-inflammatory and cadiovascular effects. In our serious of studies on the anticancer natural medicines, we have found some active compounds. As a continuation of our work for discovering more effective components, we have now investigated G. pentaphyllum (Thunb.) Makino, which is the first example of ginsenosides (Rb1, Rb3, Rd, etc.) ever found from a plant not belonging to the Araliaceae. Based on the review on the chemical constituents and pharmacological research of G. pentaphyllum, the chemical constituents were studied and their antitumor activities were tested.
By means of many different chromatographic methods such as silica gel, Sephadex LH-20, reversed phase ODS column and P-HPLC chromatography, 38 compounds were isolated from the 75% ethanol extract of the herbs of Gynostemma pentaphyllum. On the basis of spectroscopic analysis (UV, IR, NMR, MS) and physicochemical characters, they were identified as (23S)-21R-O-n-butyl-3β,20ξ,21-trihydroxy-21,23-epoxydammar-24-ene 3-O-[a-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (1), (23S)-21S-O-n-butyl-3β,20ξ,21 -trihydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl-(1→2)][β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (2), (23S)-21R-O-n-butyl-19-oxo-3β,20ξ,21 -trihydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rharnnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (3), (23S)-21S-O-n-butyl-19-oxo-3β,20ξ,21 -trihydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyran-osyl(1→3)]-α-L-arabinopyranoside (4), (23S-21R-O-n-butyl-3β,20ξ,21 -trihydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (5), (20S)-3β,20,21-trihydroxydammar-24-ene 3-O-[α-L-rhamnopyranosyl-(1→2)][β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (6), gylongiposideⅠ(7),α-spinaster-ol (8),β-sitosterol (9),α-spinasterol 3-Oβ-β-D-glucopyranoside (10), daucosterol (11), gypenosideⅢ(ginsenoside Rb1, 12), gypenosideⅣ(ginsenoside Rb3, 13), gypenosideⅧ(ginsenoside Rd, 14), gypenosideⅪⅤ(15), gypenosideⅫ(ginsenoside F2,16), gypenosideⅩⅢ(17), rutin (18), malonic acid (19), (20R,23R)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-0-[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-6-O- acetyl-β-D-glucopyranoside (20), (20S,23S)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-6-O-acetyl-β-D-glucopyranoside (21), gypenoside XLIX (22), (20S)-3β,20,21-trihydroxydarnmar-24-ene 3-O- {[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-β-D-glucopyranosyl}-21 -O-β-D-glucopyranoside (23), (23S,23S)-19-oxo-3β,20ξ,21,26-tetrahydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (24), (23R,23R)-19-oxo-3β,20ξ,21,26-tetrahydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (25), (20R,23R)-19-oxo-3β,20-dihydroxydammar-24-en-21-oci acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl-(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (26), (20S,23S)-19-oxo-3β,20-dihydroxydammar-24-en-21-oci acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (27), (205)-3β,20,21-trihydroxydam-mar-23,25-diene 3-0-{[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)] -β-D-glucopyranosyl}-21-O-β-D)-glucopyranoside (28), (205)-19-oxo-3β,20,21 -trihydroxydam-mar-23,25-diene 3-O-{[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-a-L-arabinopyranosyl}-21-O-β-D-glucopyranoside (29), (23S)-21R-O-ethyl-19-oxo-3β,20ξ,21-trihydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyran-osyl(1→3)]-α-L-arabinopyranoside (30), (235)-19-oxo-3β,20ξ,21ξ-trihydroxy-21,23-epoxyda-mmar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(13)]-α-L-arabino-pyranoside (31), (20S)-19-oxo-3β,20,21ξ,25-tetrahydroxy-21,24ξ-cyclodammarane 3-0-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (32), (20R)-19-oxo-3β,20,21ξ,23ξ-tetrahydroxy-21,24ξ-cyclodammar-24-ene 3-O-[α-L-rham-nopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (33), (20R,23R)-3β, 20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (34), (205,235)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl-(1→3)]-β-D-glucopyranoside (35), (205,235)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (36), (20R,23R)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (37), quercetin (38). Among them, compounds 1-6, 24-29, 36 and 37 were determined as new compounds by chemical and spectroscopic methods.
In the course of our search for triterpene saponins with antitumor activity, by MTT methods, the in vitro cytotoxicity activities against cancer cell lines of some compounds isolated were assayed. The effects of compounds on human cancer cells growth, expressed as the IC50 value, for HL-60 cells lines compounds 1-5,26,33,36-37 showed growth inhibitory effects and differentiation induction abilities, compound 3 was the most potent with the IC50 value of 8.45μmol/L. For Colon 205 and Du 145 cell lines 1-5, and 7 showed growth inhibitory effects and differentiation induction abilities, compound 2 showed strong against Colon 205 cell lines with IC50 value of 22.95μmol/L, and compound 3 showed strong against Du 145 cell lines with IC50 value of 17.41μmol/L. For A2780 and OVCAR-3 cells lines compounds 20,21,34 and 35 showed growth inhibitory effects and differentiation induction abilities, compound 20 showed moderate antitumor activities against A2780 and OVCAR-3 cell lines with IC50 value of 10.40μmol/L and 2.8μmol/L, respectively.
本文在综述了绞股蓝的化学成分和药理活性研究的基础上,对其进行了系统的化学成分研究及抗肿瘤活性研究。采用反复硅胶柱色谱法、大孔树脂、Sephadex LH-20、反相ODS、半制备型HPLC等多种色谱手段从绞股蓝的75%乙醇提取物中,分离得到38个化合物。并通过理化性质、波谱数据分析(IR、NMR、MS等)和化学方法鉴定了它们的结构,分别为：(23S)-21R-O-正丁基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-比喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(1),(23S)-21S-O-正丁基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(2),(23S)-21R-O-正丁基-19-醛基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(3),(23S)-21S-O-正丁基-19-醛基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(4),(23S)-21R-O-正丁基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(5),(20S)-3β,20,21-三羟基达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(6),长梗绞股蓝皂苷Ⅰ(7),α-菠菜甾醇(8),β-谷甾醇(9),α-菠菜甾醇3-O-β-D-吡喃葡萄糖苷(10),胡萝卜苷(11),绞股蓝皂苷Ⅲ(人参皂苷Rb1,12),绞股蓝皂苷Ⅳ(人参皂苷Rb3,13),绞股蓝皂苷Ⅷ(人参皂苷Rd,14),绞股蓝皂苷ⅩⅣ(15),绞股蓝皂苷Ⅻ(人参皂苷F2,16),绞股蓝皂苷ⅩⅢ(17),芸香苷(18),丙二酸(19),(20R,23R)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-6-O-乙基-β-D-吡喃葡萄糖苷(20),(20S,23S)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-6-O-乙基-β-D-吡喃葡萄糖苷(21),绞股蓝皂苷XLIX (22),(20S)-3β,20,21-三羟基达玛烷-24-烯3-O-{[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖基}-21-O-β-D-吡喃葡萄糖苷(23),(23S,23S)-19-醛基-3β,20ξ,21,26-四羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(24),(23R,23R)-19-醛基-3β,20ξ,21,26-四羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(25),(20R,23R)-19-醛基-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(26),(20S,23S)-19-醛基-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(27),(20S)-3β,20,21-三羟基达玛-23,25-二烯3-O-{[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖基}-21-O-β-D-吡喃葡萄糖苷(28),(20S)-19-醛基-3β,20,21-三羟基达玛-23,25-二烯3-O-{[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖基}-21-O-β-D-吡喃葡萄糖苷(29),(23S)-21R-O-乙基-19-醛基-3β,20ξ,21-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(30),(23S)-19-醛基-3β,20ξ,21ξ-三羟基-21,23-环氧达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(31),(20S)-19-醛基-3β,20,21ξ,25-四羟基-21,24ξ-环达玛烷3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(32),(20R)-19-醛基-3β,20,21ξ,23ξ-四羟基-21,24ξ-环达玛-24-烯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷(33),(20R,23R)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(34),(20S,23S)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[α-L-吡喃鼠李糖基(1→2)][β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(35),(20S,23S)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[β-D-吡喃木糖基(1→3)]-β-D-吡喃葡萄糖苷(36), (20R,23R)-3β,20-二羟基达玛-24-烯-21-羧酸21,23-内酯3-O-[β-D-吡喃木糖基(1→3)][β-D-吡喃葡萄糖苷(37),槲皮素(38)。其中未见文献报道的新化合物14个,分别为化合物1-6、24-29、36和37。
采用MTT法,对从绞股蓝中得到的部分化合物的体外肿瘤抑制活性进行了初步筛选和评价。化合物1-5,26,33,36-37对细胞株HL-60均具有不同程度的细胞生长抑制作用,其中化合物3活性最强,IC50值为8.45μmol/L.化合物1-5,7对细胞株Colon205和Du145不同浓度下均具有生长抑制作用,其中化合物2对Colon 205的活性最强,IC50值为22.95μmol/L,化合物3对Du145的活性最强,IC50值为17.41μmol/L。化合物20-21,34-35对细胞株A2780和OVCAR-3在不同浓度下均具有生长抑制作用,其中化合物20和34对A2780有较强的活性,其IC50均为10.4μmol/L,化合物20对OVCAR-3有较强的活性,其IC50为2.8μmol/L。
Gynostemma pentaphyllum (Thunb.) Makino (Cucurbitaceae) is a herbal medicine with anticancer activity, widely distributed in China, Korea and Japan. It is famous as "Southen Ginseng". The biological active constituents are dammarane-type glycosides, called gypenosides, which are structurally correlated to the ginseng saponins. Pharmacological investigation demonstrated that extracts of this plant exhibited a variety of biological effects, such as anticancer, anti-inflammatory and cadiovascular effects. In our serious of studies on the anticancer natural medicines, we have found some active compounds. As a continuation of our work for discovering more effective components, we have now investigated G. pentaphyllum (Thunb.) Makino, which is the first example of ginsenosides (Rb1, Rb3, Rd, etc.) ever found from a plant not belonging to the Araliaceae. Based on the review on the chemical constituents and pharmacological research of G. pentaphyllum, the chemical constituents were studied and their antitumor activities were tested.
By means of many different chromatographic methods such as silica gel, Sephadex LH-20, reversed phase ODS column and P-HPLC chromatography, 38 compounds were isolated from the 75% ethanol extract of the herbs of Gynostemma pentaphyllum. On the basis of spectroscopic analysis (UV, IR, NMR, MS) and physicochemical characters, they were identified as (23S)-21R-O-n-butyl-3β,20ξ,21-trihydroxy-21,23-epoxydammar-24-ene 3-O-[a-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (1), (23S)-21S-O-n-butyl-3β,20ξ,21 -trihydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl-(1→2)][β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (2), (23S)-21R-O-n-butyl-19-oxo-3β,20ξ,21 -trihydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rharnnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (3), (23S)-21S-O-n-butyl-19-oxo-3β,20ξ,21 -trihydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyran-osyl(1→3)]-α-L-arabinopyranoside (4), (23S-21R-O-n-butyl-3β,20ξ,21 -trihydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (5), (20S)-3β,20,21-trihydroxydammar-24-ene 3-O-[α-L-rhamnopyranosyl-(1→2)][β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (6), gylongiposideⅠ(7),α-spinaster-ol (8),β-sitosterol (9),α-spinasterol 3-Oβ-β-D-glucopyranoside (10), daucosterol (11), gypenosideⅢ(ginsenoside Rb1, 12), gypenosideⅣ(ginsenoside Rb3, 13), gypenosideⅧ(ginsenoside Rd, 14), gypenosideⅪⅤ(15), gypenosideⅫ(ginsenoside F2,16), gypenosideⅩⅢ(17), rutin (18), malonic acid (19), (20R,23R)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-0-[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-6-O- acetyl-β-D-glucopyranoside (20), (20S,23S)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-6-O-acetyl-β-D-glucopyranoside (21), gypenoside XLIX (22), (20S)-3β,20,21-trihydroxydarnmar-24-ene 3-O- {[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-β-D-glucopyranosyl}-21 -O-β-D-glucopyranoside (23), (23S,23S)-19-oxo-3β,20ξ,21,26-tetrahydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (24), (23R,23R)-19-oxo-3β,20ξ,21,26-tetrahydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (25), (20R,23R)-19-oxo-3β,20-dihydroxydammar-24-en-21-oci acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl-(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (26), (20S,23S)-19-oxo-3β,20-dihydroxydammar-24-en-21-oci acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (27), (205)-3β,20,21-trihydroxydam-mar-23,25-diene 3-0-{[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)] -β-D-glucopyranosyl}-21-O-β-D)-glucopyranoside (28), (205)-19-oxo-3β,20,21 -trihydroxydam-mar-23,25-diene 3-O-{[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-a-L-arabinopyranosyl}-21-O-β-D-glucopyranoside (29), (23S)-21R-O-ethyl-19-oxo-3β,20ξ,21-trihydroxy-21,23-epoxydammar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyran-osyl(1→3)]-α-L-arabinopyranoside (30), (235)-19-oxo-3β,20ξ,21ξ-trihydroxy-21,23-epoxyda-mmar-24-ene 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(13)]-α-L-arabino-pyranoside (31), (20S)-19-oxo-3β,20,21ξ,25-tetrahydroxy-21,24ξ-cyclodammarane 3-0-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (32), (20R)-19-oxo-3β,20,21ξ,23ξ-tetrahydroxy-21,24ξ-cyclodammar-24-ene 3-O-[α-L-rham-nopyranosyl(1→2)][β-D-xylopyranosyl(1→3)]-α-L-arabinopyranoside (33), (20R,23R)-3β, 20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl(1→2)] [β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (34), (205,235)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[α-L-rhamnopyranosyl(1→2)][β-D-xylopyranosyl-(1→3)]-β-D-glucopyranoside (35), (205,235)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (36), (20R,23R)-3β,20-dihydroxydammar-24-en-21-oic acid 21,23-lactone 3-O-[β-D-xylopyranosyl(1→3)]-β-D-glucopyranoside (37), quercetin (38). Among them, compounds 1-6, 24-29, 36 and 37 were determined as new compounds by chemical and spectroscopic methods.
In the course of our search for triterpene saponins with antitumor activity, by MTT methods, the in vitro cytotoxicity activities against cancer cell lines of some compounds isolated were assayed. The effects of compounds on human cancer cells growth, expressed as the IC50 value, for HL-60 cells lines compounds 1-5,26,33,36-37 showed growth inhibitory effects and differentiation induction abilities, compound 3 was the most potent with the IC50 value of 8.45μmol/L. For Colon 205 and Du 145 cell lines 1-5, and 7 showed growth inhibitory effects and differentiation induction abilities, compound 2 showed strong against Colon 205 cell lines with IC50 value of 22.95μmol/L, and compound 3 showed strong against Du 145 cell lines with IC50 value of 17.41μmol/L. For A2780 and OVCAR-3 cells lines compounds 20,21,34 and 35 showed growth inhibitory effects and differentiation induction abilities, compound 20 showed moderate antitumor activities against A2780 and OVCAR-3 cell lines with IC50 value of 10.40μmol/L and 2.8μmol/L, respectively.
引文
[1]陈书坤.绞股蓝属植物的分类系统和分布[J].植物分类学报,1995,33(4)：403-410.
[2]中华本草[M].上海科技出版社,2000,5.532.
[3]陈建国,中草药,1990,21(9)：40.
[4]门田晓美,公开特许公报,1986,昭61-265065.
[5]Nagai M., et al.. Abstracts, the 23rd Meeting of the Japanese Society of Pharmacognosy, Hiroshima, 1976,11:37.
[6]Takemoto T., Arihara S., Nakajima T., Okuhira M.. Studies on the Constituents of Gynostemma pentaphyllum Makino. I. Structures of Gypenoide I-XIV [J]. Yakugaku Zassh,1983,103(2):173-185.
[7]Takemoto T., Arihara S., Nakajima T., Okuhira M.. Studies on the Constituents of Gynostemma pentaphyllum Makino. Ⅱ. Structures of Gypenoide XV-XXI [J]. Yakugaku Zassh,1983,103(10): 1015-1023.
[8]Takemoto T., Arihara S., Yoshikawa K., Nakajima T., and Okuhira M.. Studies on the Constituents of Cucurbitaceae Plants. VII. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (3) [J]. Yakugaku Zassh,1984,104(4):325-331.
[9]Takemoto T., Arihara S., Yoshikawa K., Nakajima T., and Okuhira M.. Studies on the Constituents of Cucurbitaceae Plants. VIII. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (4) [J]. Yakugaku Zassh,1984,104(4):332-339.
[10]Takemoto T., Arihara S., Yoshikawa K., Hino K., Nakajima T., Okuhira M.. Studies on the Constituents of Cucurbitaceae Plants. IX. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (5) [J]. Yakugaku Zassh,1984,104(7):724-730.
[11]Takemoto T., Arihara S., Yoshikawa K., Hino K., Nakajima T., Okuhira M.. Studies on the Constituents of Cucurbitaceae Plants. X. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (6) [J]. Yakugaku Zassh,1984,104(9):939-945.
[12]Takemoto T., Arihara S., Yoshikawa K., Hino K., Nakajima T., Okuhira M.. Studies on the Constituents of Cucurbitaceae Plants. XI. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (7) [J]. Yakugaku Zassh,1984,104(10):1043-1049.
[13]Takemoto T., Arihara S., Yoshikawa K., Hino K., Nakajima T., Okuhira M.. Studies on the Constituents of Cucurbitaceae Plants. Ⅻ. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (8) [J]. Yakugaku Zassh,1984,104(11):1155-1162.
[14]Takemoto T., Arihara S., and Yoshikawa K.. Studies on the Constituents of Cucurbitaceae Plants. XIV On the Saponin Constituents of Gynostemma pentaphyllum Makino. (9) [J]. Yakugaku 5Zassh,1986, 106(8):664-670.
[15]Yoshikawa K., Takemoto T., Arihara S.. Studies on the Constituents of Cucurbitaceae Plants. XV. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (10) [J]. Yakugaku Zassh,1986,106(9): 758-763.
[16]Yoshikawa K., Arimitsu M., and Arihara S.. Studies on the Constituents of Cucurbitaceae Plants. XVI. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (11) [J]. Yakugaku Zassh,1987, 107(4):262-267.
[17]Yoshikawa K., Mitake M., Takemoto T., and Arihara S.. Studies on the Constituents of Cucurbitaceae Plants. XVII. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (12) [J]. Yakugaku Zassh,1987,107(5):355-360.
[18]Yoshikawa K., Arimitsu M., Kishi K., Takemoto T., and Arihara S.. Studies on the Constituents of Cucurbitaceae Plants. XVIII. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (13) [J]. Yakugaku Zassh,1987,107(5):361-366.
[19]Nagai M., Izawa K., Nagumo S., Sakurai N., and Inoue T.. Two Glycosides of a Novel Dammarane Alcohol from Gynostemma pentaphyllum [J]. Chem Pharm Bull,1981,29(3):779-783.
[20]Kuwahara M., Kawanishi F., Komiya T., and Oshio H.. Dammarane Saponins of Gynostemma pentaphyllum Makino and Isolation Malonylginsenosides-Rb1,-Rd, and Malonylgypenoside V [J]. Chem. Pharm. Bull.,1989,37(1):135-139.
[21]吴基良,邱培伦.绞股蓝化学成分研究进展[J].西北药学杂志,1990,5(2)：31-35.
[22]覃章铮,赵蕾,毕世荣,游兰.绞股蓝的皂甙成份及资源[J].天然产物研究与开发,1992,4(1)：83-98.
[23]刘欣.三种常用中药的化学成分研究.中国药科大学博士学位论文,2002.
[24]沈宏伟,肖彦春,车仁国,金凤燮,鱼红闪.绞股蓝化学成分研究现状[J].时珍国医国药,2008,19(7)：1561-1564.
[25]Razmovski-Naumovski V., Huang T. H. W., Tran V. H., Li Q., Duke C. C., and Roufogalis B. D.. Chemistry and pharmacology of Gynostemma pentaphyllum [J]. Phytochemistry Reviews,2005,4: 197-219.
[26]郭绪林,王铁军,边宝林.长梗绞股蓝的化学成分研究[J].药学学报,1997,32(7)：524-529.
[27]Yin F., Hu L. H., Pan R. X.. Novel Dammarane-Type Glycosides from Gynostemma pentaphyllum [J]. Chem. Pharm. Bull.,2004,52(12):1440-1444.
[28]Hu L. H., Chen Z. L., Xie Y. Y.. New Triterpenoid Saponins from Gynostemma pentaphyllum [J]. J. Nat. Prod.,1996,59(12),1143-1145.
[29]Yoshikawa K., Arihara S., Matsuura K. and Miyase T.. Dammarane Saponins from Gymnema sylvestre [J]. Phytochemistry,1992,31(1):237-241.
[30]Yin F., Hu L. H., Lou F. C. and Pan R. X.. Dammarane-Type Glycosides from Gynostemma pentaphyllum [J]. J. Nat. Prod.,2004,67:942-952.
[31]Hu L. H., Chen Z. L., Xie Y. Y.. Dammarane-Type Glycosides from Gynostemma pentaphyllum [J]. Phytochemistry,1997,44(4):667-670.
[32]刘欣,叶文才,萧文鸾,车镇涛,赵守训.绞股蓝的化学成分研究[J].中国药科大学学报,2003,34(1)： 21-24.
[33]Hung T. M., Thu C. V., Cuong T. D., et al. Dammarane-Type Glycosides from Gynostemma pentaphyllum and Their Effects on IL-4-Induced Eotaxin Expression in Human Bronchial Epithelial Cells [J]. J. Nat. Prod.,2010,73,192-196.
[34]Piacente S. and Pizza C., New Dammarane-Type Glycosides from Gynostemma pentaphyllum [J]. J. Nat. Prod.,1995,58(4),512-519.
[35]Huang T. H.-W., Razmovski-Naumovski V., Salam N. K., Roufogalis B. D., et al. A Novel LXR-a activator identified from the Natural Product Gynostemma pentaphyllum [J]. Biochemical Pharmacology, 2005,70(9):1298-1308.
[36]Yin F., Hu L. H., Six New Triterpene Saponins with a 21,23-Lactone Skeleton from Gynostemma pentaphyllum [J]. Helvetica Chimica Acta,2005,88(5):1126-1134.
[37]Fang Z. P., Zeng X. Y.. Structure of gypentonoside from Gynostemma pentaphyllum Makino [J]. Acta Pharmaceutica Sinica,1996,31(9):680-683.
[38]Liu X., Ye W. C., Mo Z. Y., et al. Five New Ocotillone-Type Saponins from Gynostemma pentaphyllum [J]. J. Nat. Prod.,2004,67:1147-1151.
[39]Liu X., Yu R. M., Hsiao W. L., Zhao S. X. and Ye W. C.. Three New Dammarane Glycosides from Gynostemma pentaphyllum [J]. Chinese Chemical Letters,2004,15(1):46-48.
[40]Yin F., Zhang Y. N., Yang Z. Y. and Hu L. H.. Nine New Dammarane Saponins from Gynostemma pentaphyllum [J]. Chemistry & Biodiversity,2006,3:771-782.
[41]Norberg A., Hoa N. K., Liepinsh E., et al. A Novel Insulin-releasing Substance, Phanoside, from the Plant Gynostemma pentaphyllum [J]. The Journal of Biological Chemistry,2004,279(40):41361-41367.
[42]Zhang Z., Zhang W., Ji Y. P., et al. Gynostemosides A-E,megastigmane glycosides from Gynostemma pentaphyllum [J]. Phytochemistry,2010,71:693-700.
[43]Akihisa T., Ghosh P.,Thakur P., Rosenstein F. U. and Matsumoto T. J. Am. Oil Chem. Soc.,1986,63: 653.
[44]Akihisa T., Shimizu N., Tamura T. and Matsumoto T. Lipids,1986,21:515.
[45]Akihisa T., Kanari M., Tamura T. and Matsumoto T.. (24R)-and (24S)-14a-methyl-5a-ergost-9(11)-en-3β-ols from Gynostemma pentaphyllum [J]. Phytochemistry,1989,28(4):1271-1273.
[46]Akihisa T., Mihara H., Fujikawa T., Tamura T. and Matsumoto T..24,24-dimethyl-5a-cholestan-3β-ol, a sterol from Gynostemma pentaphyllum [J]. Phytochemistry,1988,27(9):2931-2933.
[47]Akihisa T., Tamura T., Matsumoto T., Kokke W. C. M. C., and Yokota T. Isolation of Acetylenic Sterols from a Higher Plant. Further Evidence That Marine Sterols Are Not Unique [J]. J. Org. Chem., 1989,54(3):606-610.
[48]Akihisa T., Tamura T., and Matsumoto T..14a-methyl-5a-ergosta-9(11),24(28)-dien-3(3-ol, A Sterol from Gynostemma pentaphyllum [J]. Phytochemistry,1987,26(8):2412-2413.
[49]Akihisa T., Kokke W. C. M. C., Yokota T., Tamura T., and Matsumoto T..4a,14a-dimethyl-5a-ergosta-7,9(11),24(28)-trien-3β-ol, from Phaseolus vulgaris and Gynostemma pentaphyllum [J]. Phytochemistry,1990,29(5):1647-1651.
[50]Akihisa T., Thakur S., Rosenstein F. U. and Matsumoto T.. Lipids,1986,21:39.
[51]Marino A., Elberti M. G, Cataldo A., et al. Sterols from Gynostemma pentaphyllum [J]. Soc. Ital. Biol. Sper.,1989,65(4):317-319.
[52]方乍浦,曾宪仪,等.绞股蓝中黄酮甙,有机酸的分离和鉴定[J].中国中药杂志,1989,14(11)：676-678.
[53]魏均娴,陈业高,徐兴伦,等.绞股蓝的化学成分研究Ⅰ.一种新的达玛烷—绞股蓝皂苷元Ⅱ的结构[J].化学学报,1991,49(9)：932-936.
[54]宋淑亮,吉爱国,梁浩,王伟莉.绞股蓝多糖的分离纯化及初步结构分析[J].中药材,2008,31(3)：454-456.
[55]宋淑亮,唐金宝,吉爱国,梁浩,朱鹏,王伟莉.绞股蓝多糖的提取纯化和含量测定[J].中药材,2006,29(6)：595-597.
[56]王绍晶,罗巅辉.碱提绞股蓝水溶性多糖的研究[J].食品研究与开发,2006,27(5)：92-94.
[57]邓世林,李新凤,陈本美,等.绞股蓝中氨基酸、维生素及多种化学元素的分析[J].湖南医科大学学报,1994,19(6)：487-490.
[58]陈业高,张燕,等.绞股蓝中苯甲醇葡萄糖甙的分离和鉴定[J].云南师范大学学报,2001,21(2)：58-59.
[59]Sugimori T., Tsukada Y., Taki H.. Preparation of Phyllosultion [J]. Japan Kokai,1975,75:112,400.
[60]竹本常松,等.公开特许公报,昭58-59921.
[61]王玉琴,等.绞股蓝总皂甙的抗肿瘤作用[J].中西医结合杂志,1988,8(5)：286.
[62]徐长福,王冰,任淑婷,张健,等.绞股蓝总皂甙对小鼠S180肉瘤及K562细胞的抑制作用[J].中西安医科大学学报,2002,23(6)：217-219.
[63]刘华,司履生.绞股蓝总皂甙对体外培养肺癌细胞的抑制作用[J].西安医科大学学报,1994,15(4)：346-348.
[64]刘侠,汪平君,许铰新.绞股蓝总皂苷抑制小鼠Lewis肺癌生长与提高免疫力研究[J].安徽中医学院学报,2001,20(1)：43-44.
[65]冯国宜.抗癌植物资源与天然产物研究[J].湖北民族学院学报(自然科学版),2001,8(3)26-32.
[66]Lu H. F., Chen Y. S., Yang J. S., et al. Gypenoside induced G0/G1 arrest via inhibition of cyclin E and induction of apoptosis via activation of caspase-3 and-9 in human lung cancer A-549 cells [J]. In Vivo 2008,22:215-221.
[67]Chen J. C., Lu K. W., Tsai M. L., et al. Gypenosides induced G0/G1 arrest via CHk2 and apoptosis through endoplasmic reticulum stress and mitochondria-dependent pathways in human tongue cancer SCC-4 cells [J]. Oral Oncol.2009,45:273-283.
[68]辛冬生,邱培伦,刘俊田,等.绞股蓝总皂苷对实验性心肌缺血缺氧损伤的保护作用[J].中国药学杂志,1990,25(7)：398-401.
[69]张芳,等.绞股蓝皂甙对实验性大鼠心肌缺血、心脏收缩功能的保护作用[J].山东医科大学学报,1999,37(1)：31-33.
[70]Han X. Y, Wei H. B., Zhang F. C.. Analysis of the inhibitory effect of gypenoside on Na+, K+-ATPase in rats'heart and brain and its kinetics [J]. Chin. J. Integr. Med.2007,13:128-131.
[71]Huang T. H., Tran V. H., Roufogalis B. D., et al. Gypenoside XLIX, a naturally occurring PPAR-aactivator, inhibits cytokine-induced vascular cell adhesion molecule-1 expression and activity in human endothelial cells [J]. European Journal of Pharmacology,2007,565:158-165.
[72]Huang T. H., Li Y., Razmovski-Naumovsi V, et al. Gypenoside XLIX isolated from Gynostemma pentaphyllum inhibits nuclear factor-kappap activation via a PPAR-alpha-dependent pathway [J]. Journal of Biochemical Science,2006,13:535-548.
[73]池明宇,梅学文,郑贵元,等.绞股蓝丹对大鼠局灶性脑缺血再灌注损伤的保护作用[J].中医研究,2003,16(4)：18-20.
[74]史以菊,邢国庆,邱玉芳,等.绞股蓝对小鼠脑血流量及耐氧能力的影响[J].现代康复,2001,5(5)：117-119.
[75]王旭平,赵玲,冯玉新,等.绞股蓝总苷对谷氨酸诱导的胎鼠大脑皮层神经元氧化性损伤保护机制的研究[J].山东大学学报,2006,44(6)：564-566.
[76]李乐,高小利,丁宝兴.绞股蓝总黄酮对缺氧心肌细胞损伤标记物及细胞内游离Ca2+浓度的影响[J].浙江工业大学学报,2008,36(1)：23-25.
[77]葛君,朱妤捷,吴军,等.绞股蓝皂苷对家兔肠缺血再灌注致心肌损伤的保护作用[J].蚌埠医学院学报,2006,31(4)：347-350.
[78]张小丽,刘珍,朱自平,等.绞股蓝总苷对体内外血栓及凝血功能的影响[J].华西药学杂志,1999,14(5)：335-337.
[79]池明宇,张澄波,郑贵元,等.复方绞股蓝对大鼠实验性体内血栓形成的研究[J].中医药学刊,2003,21(12)：1998-2000.
[80]董晓晖.绞股蓝总苷与乙酰水杨酸对血小板聚集和血栓形成的影响[J].辽宁医学院学报,2006,29(3)：47-48.
[81]段炳南,陈庆林.绞股蓝总皂甙对小鼠腹腔巨噬细胞内酶活性及吞噬功能的影响[J].江西医学院学报,2007,47(3)：38-39.
[82]周俐,叶开和,任先达.绞股蓝总苷对免疫功能低下小鼠模型特异性免疫功能的影响[J].中华中医药学刊,2008,26(1)：145-146.
[83]张海燕,郭强,温伟业.绞股蓝总皂苷对小鼠免疫功能的影响[J].中兽医学杂志,2006(2)：13-15.
[84]王斌,葛志东,周爱武,等.绞股蓝总皂苷体外对免疫细胞功能的影响[J].中药新药与临床药理,1999,10(1)：36-38.
[85]陈宏卫,王守丽,孟贺丽,张在和.绞股蓝皂苷对糖尿病大鼠降糖作用的观察[J].中国糖尿病杂志,1997,5(4)：229-230.
[86]魏守蓉,薛存宽,何学斌,等.绞股蓝多糖降血糖作用的实验研究[J].中国老年学杂志,2005,25(4)： 418-420.
[87]黄萍,陈竞龙,张雷,等.绞股蓝皂甙对2型糖尿病肾病的血脂、微量白蛋白尿的影响[J].中国现代医学杂志,2007,17(2)：206-207.
[88]包海花,郭新民,聂影,等.绞股蓝总皂甙对2型糖尿病大鼠脑神经生长因子基因表达的影响[J].中国康复医学杂志,2006,21(4)：328-329.
[89]谭华炳.绞股蓝对兔高脂血症和血液流变学影响研究[J].第三军医大学学报,2007,29(15)：1497-1450.
[90]谭华炳.绞股蓝预防兔动脉粥样硬化的研究[J].中国老年学杂志,2007,2(6)：519-521.
[91]谭华炳,唐中科,赵世印,等.野生绞股蓝复方治疗高脂血症临床研究[J].浙江中西医结合杂志,2004,14(7)：409-410.
[92]谭华炳.绞股蓝对高胆同醇饵料饲养兔血液流变学和CRP异常的干预研究[J].中国微循环杂志,2007,11(2)：108-110.
[93]王树,桂潘莹.复方绞股蓝胶囊对高脂血症小鼠血脂的影响[J].广西中医药,2005,28(3)54-56.
[94]田健,董晓晖,于信民,等.绞股蓝总皂苷对实验性高脂血症大鼠内皮素的影响[J].中国煤炭工业医学杂志,2005,8(8)：906-908.
[95]Lin J. M., Lin C. C., Chiu H. F., et al. Evaluation of the anti-inflammatory and liver-protective effects of Anoectochilus formosams, Ganoderma lucidum and Gynostemma pentaphyllum in rats [J]. The American Journal of Chinese Medicine,1993,21:59-69.
[96]万丽,万兴旺,胡晋红.绞股蓝总皂苷对免疫性肝纤维化大鼠肝功能和肝纤维化的影响[J].第二军医大学学报,2003,24(12)：1319-1321.
[97]陶建武,姚彩萍,王冀春,等.绞股蓝在大鼠肝的抗脂质过氧化作用[J].中国临床药学杂志,2001,10(3)：160-162.
[98]胡宝春,张金桃,李晓清,曾明新.绞股蓝总皂式对小鼠四氯化碳肝损伤肝组织一氧化和谷胱甘肽的影响[J].武警医学,1999,10(2)：71-73.
[99]张慧丽,宋有涛,辛如,等.超声提取绞股蓝总皂苷及抗氧化作用的研究[J].辽宁大学学报(自然科学版),2006,33(4)：346-348.
[100]Wang Z. J., Luo D. H.. Antioxidant activities of different fractions of polysaccharide purified from Gynostemma pentaphyllum Makino [J]. Carbohydrate Polymers,2007,68:54-58.
[101]张谟瑞,陈谦,吴宪华.多功能抗衰老药——绞股蓝[J].中医药信息,1996,13(4)：28-30.
[102]刘青青,吴景东.绞股蓝提取液对自然衰老影响的实验研究[J].辽宁中医药大学学报,2008,10(6)：203-205.
[103]刘国辉,姚丹丹,卢佳怡,等.绞股蓝对D-半乳糖所致亚急性衰老大鼠下丘脑的抗衰老作用及其机制研究[J].医学理论与实践,2006,19(5)：497-499.
[104]陈珏,许衡钧.绞股蓝对果蝇寿命的影响[J].中国药理学通报,1987,3(6)：340-342.
[105]Rujjanawate C., Kanjanapothi D., Amornlerdpison D.. The anti-gastric ulcer effect of Gynostemma pentaphyllum Makino [J]. Phytomedicine,2004,11:431-435.
[106]潘峰,刘迪,黄翠霞,薛秀琴.绞股蓝皂甙的药理与临床研究[J].现代中西医结合杂志,2006,15(5)：674-676.
[107]程小跃,姚敦武,孙兆泉.绞股蓝总皂甙镇痛作用的实验研究[J].湖南中医药学报,1999,5(10)：34-35.
[108]韩晓燕,卫洪波,张富程.绞股蓝总苷对大鼠心、脑Na+, K+-ATP酶的抑制作用及其动力学分析[J].中草药,2006,37(10)：1542-1544.
[109]喻陆,廖立生.绞股蓝皂甙与AWS减轻庆大霉素对培养肾小管上皮细胞损伤的探讨[J].第三军医大学学报,1996,18(2)：162-164.
[110]喻陆,廖立生.绞股蓝皂甙对培养肾小管细胞庆大霉素损伤防治机理的探讨[J].中华肾脏病杂志,1996,12(2)：79-81.
[111]董燕湘,董晓先,何慧华.大鼠骨髓间质干细胞用中药绞股蓝诱导为神经细胞的研究[J].中华神经科杂志,2003,36(5)：355-358.
[112]Chi A. P., Chen J. P., Wang Z. Z., et al. Morphological and structural characterization of a polysaccharide from Gynostemma pentaphyllum Makino and its anti-exercise fatigue activity [J]. Carbohydrate Polymers,2008,74:868-874.
[113]王海元,池爱平.绞股蓝多糖对运动小鼠组织抗氧化能力的影响[J].食品科学,2008,29(11)：616-618.
[114]高应东,陈武,熊筱娟,等.绞股蓝皂苷对鼠应激能力影响的效果评估[J].中国临床康复,2005,9(43)：109-111.
[115]彭跃钢,黄立武.复方绞股蓝总甙胶囊对血管性痴呆大鼠海马神经细胞凋亡的影响[J].广西医科大学学报,2005,22(3)：368-370.
[116]曾晓黎.绞股蓝的体外抗菌活性试验[J].中成药,1999,21(6)：308-311.
[117]周和平.葫芦科绞股蓝的皂甙成分与药理[J].药学学报,1988,3(6)：340-343.
[118]王本祥.现代中药药理学[M].天津科学技术出版社,1999,1227-1230.
[1]Yin F., Zhang Y. N., Yang Z. Y. and Hu L. H.. Nine New Dammarane Saponins from Gynostemma pentaphyllum. Chemistry & Biodiversity [J],2006,3:771-782.
[2]Yin F., Hu L. H., Lou F. C. and Pan R. X.. Dammarane-Type Glycosides from Gynostemma pentaphyllum. J. Nat. Prod. [J],2004,67:942-952.
[3]Iwamoto M., Fujioka T., Okabe H., Mihashi K., and Yamauchi T.. Studies on the Actinostemma lobatum MAXIM. I. Structures of Actinistemmosides A, B, C and D. Dammarane Triterpene Glycosides Isolated from the Herb [J]. Chem. Pharm. Bull.1987,35(2):553-561.
[4]沈兆邦,王成章,陈祥,在原重信.一种新型的绞股蓝皂苷[J].林产化学与工业,1993,13(4)：265-269.
[5]Yin F., Hu L. H., Six New Triterpene Saponins with a 21,23-Lactone Skeleton from Gynostemma pentaphyllum. Helvetica Chimica Acta [J].2005,88(5):1126-1134.
[6]Piacente S. and Pizza C., New Dammarane-Type Glycosides from Gynostemma pentaphyllum [J]. J. Nat. Prod.,1995,58(4),512-519.
[7]Wang J. H., Li W., Sha Y., Tezuka Y, Kadota S. and Li X.. Triterpenoid Saponins from Leaves and Stems of Panax quinquefolium L. J. Asian Nat. Prod. Res [J].2001,3,123-130.
[8]郭绪林,王铁军,边宝林.长梗绞股蓝的化学成分研究[J].药学学报,1997,32(7)：524-529.
[9]Takemoto T., Arihara S., Nakajima T., Okuhira M.. Studies on the Constituents of Gynostemma pentaphyllum Makino. I. Structures of Gypenoide I-XIV. Yakugaku Zassh [J].1983,103(2):173-185.
[10]Yoshikawa K., Takemoto T., Arihara S.. Studies on the Constituents of Cucurbitaceae Plants. XV. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (10) [J]. Yakugaku Zassh,1986,106(9): 758-763.
[11]王建忠,王锋鹏.川党参的化学成分研究[J].天然产物研究与开发,1996,8(2)：8-12.
[12]李巍,李铣.文冠果果柄的化学成分研究[J].中草药,2008,38(3)：334-337.
[13]梁龙,钟炽昌,萧淖殷.傣药“亚洛轻”化学成分研究[J].中草药,1999,25(5)：236-238.
[14]孙凯,李铣.南葶苈子的化学成分[J].沈阳药科大学学报,2003,20：419-421.
[1]Lammer C, Wagerer S, Saffrich R, et al. The cdc25B phosphatase is essential for the G2/M phase transition in human cells [J]. J Cell Sci,1998,11(16):2445-2453.
[2]赵斌,葛金芳,朱娟娟.小议在MTT法测细胞增殖抑制率中IC50的计算方法[J].安徽医药,2007,11(9)：834-835.
[3]王光兰,崔俊生.人参皂苷Rg3诱导肺癌细胞NCI-H460凋亡的机制[J].吉林大学学报,2008,34(6)： 980-985.
[4]陈迪,倪劲松.人参皂苷Rg3对乳腺癌细胞MCF-7诱导凋亡作用[J].吉林大学学报,2008,34(4)： 610-614.
[2]中华本草[M].上海科技出版社,2000,5.532.
[3]陈建国,中草药,1990,21(9)：40.
[4]门田晓美,公开特许公报,1986,昭61-265065.
[5]Nagai M., et al.. Abstracts, the 23rd Meeting of the Japanese Society of Pharmacognosy, Hiroshima, 1976,11:37.
[6]Takemoto T., Arihara S., Nakajima T., Okuhira M.. Studies on the Constituents of Gynostemma pentaphyllum Makino. I. Structures of Gypenoide I-XIV [J]. Yakugaku Zassh,1983,103(2):173-185.
[7]Takemoto T., Arihara S., Nakajima T., Okuhira M.. Studies on the Constituents of Gynostemma pentaphyllum Makino. Ⅱ. Structures of Gypenoide XV-XXI [J]. Yakugaku Zassh,1983,103(10): 1015-1023.
[8]Takemoto T., Arihara S., Yoshikawa K., Nakajima T., and Okuhira M.. Studies on the Constituents of Cucurbitaceae Plants. VII. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (3) [J]. Yakugaku Zassh,1984,104(4):325-331.
[9]Takemoto T., Arihara S., Yoshikawa K., Nakajima T., and Okuhira M.. Studies on the Constituents of Cucurbitaceae Plants. VIII. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (4) [J]. Yakugaku Zassh,1984,104(4):332-339.
[10]Takemoto T., Arihara S., Yoshikawa K., Hino K., Nakajima T., Okuhira M.. Studies on the Constituents of Cucurbitaceae Plants. IX. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (5) [J]. Yakugaku Zassh,1984,104(7):724-730.
[11]Takemoto T., Arihara S., Yoshikawa K., Hino K., Nakajima T., Okuhira M.. Studies on the Constituents of Cucurbitaceae Plants. X. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (6) [J]. Yakugaku Zassh,1984,104(9):939-945.
[12]Takemoto T., Arihara S., Yoshikawa K., Hino K., Nakajima T., Okuhira M.. Studies on the Constituents of Cucurbitaceae Plants. XI. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (7) [J]. Yakugaku Zassh,1984,104(10):1043-1049.
[13]Takemoto T., Arihara S., Yoshikawa K., Hino K., Nakajima T., Okuhira M.. Studies on the Constituents of Cucurbitaceae Plants. Ⅻ. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (8) [J]. Yakugaku Zassh,1984,104(11):1155-1162.
[14]Takemoto T., Arihara S., and Yoshikawa K.. Studies on the Constituents of Cucurbitaceae Plants. XIV On the Saponin Constituents of Gynostemma pentaphyllum Makino. (9) [J]. Yakugaku 5Zassh,1986, 106(8):664-670.
[15]Yoshikawa K., Takemoto T., Arihara S.. Studies on the Constituents of Cucurbitaceae Plants. XV. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (10) [J]. Yakugaku Zassh,1986,106(9): 758-763.
[16]Yoshikawa K., Arimitsu M., and Arihara S.. Studies on the Constituents of Cucurbitaceae Plants. XVI. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (11) [J]. Yakugaku Zassh,1987, 107(4):262-267.
[17]Yoshikawa K., Mitake M., Takemoto T., and Arihara S.. Studies on the Constituents of Cucurbitaceae Plants. XVII. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (12) [J]. Yakugaku Zassh,1987,107(5):355-360.
[18]Yoshikawa K., Arimitsu M., Kishi K., Takemoto T., and Arihara S.. Studies on the Constituents of Cucurbitaceae Plants. XVIII. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (13) [J]. Yakugaku Zassh,1987,107(5):361-366.
[19]Nagai M., Izawa K., Nagumo S., Sakurai N., and Inoue T.. Two Glycosides of a Novel Dammarane Alcohol from Gynostemma pentaphyllum [J]. Chem Pharm Bull,1981,29(3):779-783.
[20]Kuwahara M., Kawanishi F., Komiya T., and Oshio H.. Dammarane Saponins of Gynostemma pentaphyllum Makino and Isolation Malonylginsenosides-Rb1,-Rd, and Malonylgypenoside V [J]. Chem. Pharm. Bull.,1989,37(1):135-139.
[21]吴基良,邱培伦.绞股蓝化学成分研究进展[J].西北药学杂志,1990,5(2)：31-35.
[22]覃章铮,赵蕾,毕世荣,游兰.绞股蓝的皂甙成份及资源[J].天然产物研究与开发,1992,4(1)：83-98.
[23]刘欣.三种常用中药的化学成分研究.中国药科大学博士学位论文,2002.
[24]沈宏伟,肖彦春,车仁国,金凤燮,鱼红闪.绞股蓝化学成分研究现状[J].时珍国医国药,2008,19(7)：1561-1564.
[25]Razmovski-Naumovski V., Huang T. H. W., Tran V. H., Li Q., Duke C. C., and Roufogalis B. D.. Chemistry and pharmacology of Gynostemma pentaphyllum [J]. Phytochemistry Reviews,2005,4: 197-219.
[26]郭绪林,王铁军,边宝林.长梗绞股蓝的化学成分研究[J].药学学报,1997,32(7)：524-529.
[27]Yin F., Hu L. H., Pan R. X.. Novel Dammarane-Type Glycosides from Gynostemma pentaphyllum [J]. Chem. Pharm. Bull.,2004,52(12):1440-1444.
[28]Hu L. H., Chen Z. L., Xie Y. Y.. New Triterpenoid Saponins from Gynostemma pentaphyllum [J]. J. Nat. Prod.,1996,59(12),1143-1145.
[29]Yoshikawa K., Arihara S., Matsuura K. and Miyase T.. Dammarane Saponins from Gymnema sylvestre [J]. Phytochemistry,1992,31(1):237-241.
[30]Yin F., Hu L. H., Lou F. C. and Pan R. X.. Dammarane-Type Glycosides from Gynostemma pentaphyllum [J]. J. Nat. Prod.,2004,67:942-952.
[31]Hu L. H., Chen Z. L., Xie Y. Y.. Dammarane-Type Glycosides from Gynostemma pentaphyllum [J]. Phytochemistry,1997,44(4):667-670.
[32]刘欣,叶文才,萧文鸾,车镇涛,赵守训.绞股蓝的化学成分研究[J].中国药科大学学报,2003,34(1)： 21-24.
[33]Hung T. M., Thu C. V., Cuong T. D., et al. Dammarane-Type Glycosides from Gynostemma pentaphyllum and Their Effects on IL-4-Induced Eotaxin Expression in Human Bronchial Epithelial Cells [J]. J. Nat. Prod.,2010,73,192-196.
[34]Piacente S. and Pizza C., New Dammarane-Type Glycosides from Gynostemma pentaphyllum [J]. J. Nat. Prod.,1995,58(4),512-519.
[35]Huang T. H.-W., Razmovski-Naumovski V., Salam N. K., Roufogalis B. D., et al. A Novel LXR-a activator identified from the Natural Product Gynostemma pentaphyllum [J]. Biochemical Pharmacology, 2005,70(9):1298-1308.
[36]Yin F., Hu L. H., Six New Triterpene Saponins with a 21,23-Lactone Skeleton from Gynostemma pentaphyllum [J]. Helvetica Chimica Acta,2005,88(5):1126-1134.
[37]Fang Z. P., Zeng X. Y.. Structure of gypentonoside from Gynostemma pentaphyllum Makino [J]. Acta Pharmaceutica Sinica,1996,31(9):680-683.
[38]Liu X., Ye W. C., Mo Z. Y., et al. Five New Ocotillone-Type Saponins from Gynostemma pentaphyllum [J]. J. Nat. Prod.,2004,67:1147-1151.
[39]Liu X., Yu R. M., Hsiao W. L., Zhao S. X. and Ye W. C.. Three New Dammarane Glycosides from Gynostemma pentaphyllum [J]. Chinese Chemical Letters,2004,15(1):46-48.
[40]Yin F., Zhang Y. N., Yang Z. Y. and Hu L. H.. Nine New Dammarane Saponins from Gynostemma pentaphyllum [J]. Chemistry & Biodiversity,2006,3:771-782.
[41]Norberg A., Hoa N. K., Liepinsh E., et al. A Novel Insulin-releasing Substance, Phanoside, from the Plant Gynostemma pentaphyllum [J]. The Journal of Biological Chemistry,2004,279(40):41361-41367.
[42]Zhang Z., Zhang W., Ji Y. P., et al. Gynostemosides A-E,megastigmane glycosides from Gynostemma pentaphyllum [J]. Phytochemistry,2010,71:693-700.
[43]Akihisa T., Ghosh P.,Thakur P., Rosenstein F. U. and Matsumoto T. J. Am. Oil Chem. Soc.,1986,63: 653.
[44]Akihisa T., Shimizu N., Tamura T. and Matsumoto T. Lipids,1986,21:515.
[45]Akihisa T., Kanari M., Tamura T. and Matsumoto T.. (24R)-and (24S)-14a-methyl-5a-ergost-9(11)-en-3β-ols from Gynostemma pentaphyllum [J]. Phytochemistry,1989,28(4):1271-1273.
[46]Akihisa T., Mihara H., Fujikawa T., Tamura T. and Matsumoto T..24,24-dimethyl-5a-cholestan-3β-ol, a sterol from Gynostemma pentaphyllum [J]. Phytochemistry,1988,27(9):2931-2933.
[47]Akihisa T., Tamura T., Matsumoto T., Kokke W. C. M. C., and Yokota T. Isolation of Acetylenic Sterols from a Higher Plant. Further Evidence That Marine Sterols Are Not Unique [J]. J. Org. Chem., 1989,54(3):606-610.
[48]Akihisa T., Tamura T., and Matsumoto T..14a-methyl-5a-ergosta-9(11),24(28)-dien-3(3-ol, A Sterol from Gynostemma pentaphyllum [J]. Phytochemistry,1987,26(8):2412-2413.
[49]Akihisa T., Kokke W. C. M. C., Yokota T., Tamura T., and Matsumoto T..4a,14a-dimethyl-5a-ergosta-7,9(11),24(28)-trien-3β-ol, from Phaseolus vulgaris and Gynostemma pentaphyllum [J]. Phytochemistry,1990,29(5):1647-1651.
[50]Akihisa T., Thakur S., Rosenstein F. U. and Matsumoto T.. Lipids,1986,21:39.
[51]Marino A., Elberti M. G, Cataldo A., et al. Sterols from Gynostemma pentaphyllum [J]. Soc. Ital. Biol. Sper.,1989,65(4):317-319.
[52]方乍浦,曾宪仪,等.绞股蓝中黄酮甙,有机酸的分离和鉴定[J].中国中药杂志,1989,14(11)：676-678.
[53]魏均娴,陈业高,徐兴伦,等.绞股蓝的化学成分研究Ⅰ.一种新的达玛烷—绞股蓝皂苷元Ⅱ的结构[J].化学学报,1991,49(9)：932-936.
[54]宋淑亮,吉爱国,梁浩,王伟莉.绞股蓝多糖的分离纯化及初步结构分析[J].中药材,2008,31(3)：454-456.
[55]宋淑亮,唐金宝,吉爱国,梁浩,朱鹏,王伟莉.绞股蓝多糖的提取纯化和含量测定[J].中药材,2006,29(6)：595-597.
[56]王绍晶,罗巅辉.碱提绞股蓝水溶性多糖的研究[J].食品研究与开发,2006,27(5)：92-94.
[57]邓世林,李新凤,陈本美,等.绞股蓝中氨基酸、维生素及多种化学元素的分析[J].湖南医科大学学报,1994,19(6)：487-490.
[58]陈业高,张燕,等.绞股蓝中苯甲醇葡萄糖甙的分离和鉴定[J].云南师范大学学报,2001,21(2)：58-59.
[59]Sugimori T., Tsukada Y., Taki H.. Preparation of Phyllosultion [J]. Japan Kokai,1975,75:112,400.
[60]竹本常松,等.公开特许公报,昭58-59921.
[61]王玉琴,等.绞股蓝总皂甙的抗肿瘤作用[J].中西医结合杂志,1988,8(5)：286.
[62]徐长福,王冰,任淑婷,张健,等.绞股蓝总皂甙对小鼠S180肉瘤及K562细胞的抑制作用[J].中西安医科大学学报,2002,23(6)：217-219.
[63]刘华,司履生.绞股蓝总皂甙对体外培养肺癌细胞的抑制作用[J].西安医科大学学报,1994,15(4)：346-348.
[64]刘侠,汪平君,许铰新.绞股蓝总皂苷抑制小鼠Lewis肺癌生长与提高免疫力研究[J].安徽中医学院学报,2001,20(1)：43-44.
[65]冯国宜.抗癌植物资源与天然产物研究[J].湖北民族学院学报(自然科学版),2001,8(3)26-32.
[66]Lu H. F., Chen Y. S., Yang J. S., et al. Gypenoside induced G0/G1 arrest via inhibition of cyclin E and induction of apoptosis via activation of caspase-3 and-9 in human lung cancer A-549 cells [J]. In Vivo 2008,22:215-221.
[67]Chen J. C., Lu K. W., Tsai M. L., et al. Gypenosides induced G0/G1 arrest via CHk2 and apoptosis through endoplasmic reticulum stress and mitochondria-dependent pathways in human tongue cancer SCC-4 cells [J]. Oral Oncol.2009,45:273-283.
[68]辛冬生,邱培伦,刘俊田,等.绞股蓝总皂苷对实验性心肌缺血缺氧损伤的保护作用[J].中国药学杂志,1990,25(7)：398-401.
[69]张芳,等.绞股蓝皂甙对实验性大鼠心肌缺血、心脏收缩功能的保护作用[J].山东医科大学学报,1999,37(1)：31-33.
[70]Han X. Y, Wei H. B., Zhang F. C.. Analysis of the inhibitory effect of gypenoside on Na+, K+-ATPase in rats'heart and brain and its kinetics [J]. Chin. J. Integr. Med.2007,13:128-131.
[71]Huang T. H., Tran V. H., Roufogalis B. D., et al. Gypenoside XLIX, a naturally occurring PPAR-aactivator, inhibits cytokine-induced vascular cell adhesion molecule-1 expression and activity in human endothelial cells [J]. European Journal of Pharmacology,2007,565:158-165.
[72]Huang T. H., Li Y., Razmovski-Naumovsi V, et al. Gypenoside XLIX isolated from Gynostemma pentaphyllum inhibits nuclear factor-kappap activation via a PPAR-alpha-dependent pathway [J]. Journal of Biochemical Science,2006,13:535-548.
[73]池明宇,梅学文,郑贵元,等.绞股蓝丹对大鼠局灶性脑缺血再灌注损伤的保护作用[J].中医研究,2003,16(4)：18-20.
[74]史以菊,邢国庆,邱玉芳,等.绞股蓝对小鼠脑血流量及耐氧能力的影响[J].现代康复,2001,5(5)：117-119.
[75]王旭平,赵玲,冯玉新,等.绞股蓝总苷对谷氨酸诱导的胎鼠大脑皮层神经元氧化性损伤保护机制的研究[J].山东大学学报,2006,44(6)：564-566.
[76]李乐,高小利,丁宝兴.绞股蓝总黄酮对缺氧心肌细胞损伤标记物及细胞内游离Ca2+浓度的影响[J].浙江工业大学学报,2008,36(1)：23-25.
[77]葛君,朱妤捷,吴军,等.绞股蓝皂苷对家兔肠缺血再灌注致心肌损伤的保护作用[J].蚌埠医学院学报,2006,31(4)：347-350.
[78]张小丽,刘珍,朱自平,等.绞股蓝总苷对体内外血栓及凝血功能的影响[J].华西药学杂志,1999,14(5)：335-337.
[79]池明宇,张澄波,郑贵元,等.复方绞股蓝对大鼠实验性体内血栓形成的研究[J].中医药学刊,2003,21(12)：1998-2000.
[80]董晓晖.绞股蓝总苷与乙酰水杨酸对血小板聚集和血栓形成的影响[J].辽宁医学院学报,2006,29(3)：47-48.
[81]段炳南,陈庆林.绞股蓝总皂甙对小鼠腹腔巨噬细胞内酶活性及吞噬功能的影响[J].江西医学院学报,2007,47(3)：38-39.
[82]周俐,叶开和,任先达.绞股蓝总苷对免疫功能低下小鼠模型特异性免疫功能的影响[J].中华中医药学刊,2008,26(1)：145-146.
[83]张海燕,郭强,温伟业.绞股蓝总皂苷对小鼠免疫功能的影响[J].中兽医学杂志,2006(2)：13-15.
[84]王斌,葛志东,周爱武,等.绞股蓝总皂苷体外对免疫细胞功能的影响[J].中药新药与临床药理,1999,10(1)：36-38.
[85]陈宏卫,王守丽,孟贺丽,张在和.绞股蓝皂苷对糖尿病大鼠降糖作用的观察[J].中国糖尿病杂志,1997,5(4)：229-230.
[86]魏守蓉,薛存宽,何学斌,等.绞股蓝多糖降血糖作用的实验研究[J].中国老年学杂志,2005,25(4)： 418-420.
[87]黄萍,陈竞龙,张雷,等.绞股蓝皂甙对2型糖尿病肾病的血脂、微量白蛋白尿的影响[J].中国现代医学杂志,2007,17(2)：206-207.
[88]包海花,郭新民,聂影,等.绞股蓝总皂甙对2型糖尿病大鼠脑神经生长因子基因表达的影响[J].中国康复医学杂志,2006,21(4)：328-329.
[89]谭华炳.绞股蓝对兔高脂血症和血液流变学影响研究[J].第三军医大学学报,2007,29(15)：1497-1450.
[90]谭华炳.绞股蓝预防兔动脉粥样硬化的研究[J].中国老年学杂志,2007,2(6)：519-521.
[91]谭华炳,唐中科,赵世印,等.野生绞股蓝复方治疗高脂血症临床研究[J].浙江中西医结合杂志,2004,14(7)：409-410.
[92]谭华炳.绞股蓝对高胆同醇饵料饲养兔血液流变学和CRP异常的干预研究[J].中国微循环杂志,2007,11(2)：108-110.
[93]王树,桂潘莹.复方绞股蓝胶囊对高脂血症小鼠血脂的影响[J].广西中医药,2005,28(3)54-56.
[94]田健,董晓晖,于信民,等.绞股蓝总皂苷对实验性高脂血症大鼠内皮素的影响[J].中国煤炭工业医学杂志,2005,8(8)：906-908.
[95]Lin J. M., Lin C. C., Chiu H. F., et al. Evaluation of the anti-inflammatory and liver-protective effects of Anoectochilus formosams, Ganoderma lucidum and Gynostemma pentaphyllum in rats [J]. The American Journal of Chinese Medicine,1993,21:59-69.
[96]万丽,万兴旺,胡晋红.绞股蓝总皂苷对免疫性肝纤维化大鼠肝功能和肝纤维化的影响[J].第二军医大学学报,2003,24(12)：1319-1321.
[97]陶建武,姚彩萍,王冀春,等.绞股蓝在大鼠肝的抗脂质过氧化作用[J].中国临床药学杂志,2001,10(3)：160-162.
[98]胡宝春,张金桃,李晓清,曾明新.绞股蓝总皂式对小鼠四氯化碳肝损伤肝组织一氧化和谷胱甘肽的影响[J].武警医学,1999,10(2)：71-73.
[99]张慧丽,宋有涛,辛如,等.超声提取绞股蓝总皂苷及抗氧化作用的研究[J].辽宁大学学报(自然科学版),2006,33(4)：346-348.
[100]Wang Z. J., Luo D. H.. Antioxidant activities of different fractions of polysaccharide purified from Gynostemma pentaphyllum Makino [J]. Carbohydrate Polymers,2007,68:54-58.
[101]张谟瑞,陈谦,吴宪华.多功能抗衰老药——绞股蓝[J].中医药信息,1996,13(4)：28-30.
[102]刘青青,吴景东.绞股蓝提取液对自然衰老影响的实验研究[J].辽宁中医药大学学报,2008,10(6)：203-205.
[103]刘国辉,姚丹丹,卢佳怡,等.绞股蓝对D-半乳糖所致亚急性衰老大鼠下丘脑的抗衰老作用及其机制研究[J].医学理论与实践,2006,19(5)：497-499.
[104]陈珏,许衡钧.绞股蓝对果蝇寿命的影响[J].中国药理学通报,1987,3(6)：340-342.
[105]Rujjanawate C., Kanjanapothi D., Amornlerdpison D.. The anti-gastric ulcer effect of Gynostemma pentaphyllum Makino [J]. Phytomedicine,2004,11:431-435.
[106]潘峰,刘迪,黄翠霞,薛秀琴.绞股蓝皂甙的药理与临床研究[J].现代中西医结合杂志,2006,15(5)：674-676.
[107]程小跃,姚敦武,孙兆泉.绞股蓝总皂甙镇痛作用的实验研究[J].湖南中医药学报,1999,5(10)：34-35.
[108]韩晓燕,卫洪波,张富程.绞股蓝总苷对大鼠心、脑Na+, K+-ATP酶的抑制作用及其动力学分析[J].中草药,2006,37(10)：1542-1544.
[109]喻陆,廖立生.绞股蓝皂甙与AWS减轻庆大霉素对培养肾小管上皮细胞损伤的探讨[J].第三军医大学学报,1996,18(2)：162-164.
[110]喻陆,廖立生.绞股蓝皂甙对培养肾小管细胞庆大霉素损伤防治机理的探讨[J].中华肾脏病杂志,1996,12(2)：79-81.
[111]董燕湘,董晓先,何慧华.大鼠骨髓间质干细胞用中药绞股蓝诱导为神经细胞的研究[J].中华神经科杂志,2003,36(5)：355-358.
[112]Chi A. P., Chen J. P., Wang Z. Z., et al. Morphological and structural characterization of a polysaccharide from Gynostemma pentaphyllum Makino and its anti-exercise fatigue activity [J]. Carbohydrate Polymers,2008,74:868-874.
[113]王海元,池爱平.绞股蓝多糖对运动小鼠组织抗氧化能力的影响[J].食品科学,2008,29(11)：616-618.
[114]高应东,陈武,熊筱娟,等.绞股蓝皂苷对鼠应激能力影响的效果评估[J].中国临床康复,2005,9(43)：109-111.
[115]彭跃钢,黄立武.复方绞股蓝总甙胶囊对血管性痴呆大鼠海马神经细胞凋亡的影响[J].广西医科大学学报,2005,22(3)：368-370.
[116]曾晓黎.绞股蓝的体外抗菌活性试验[J].中成药,1999,21(6)：308-311.
[117]周和平.葫芦科绞股蓝的皂甙成分与药理[J].药学学报,1988,3(6)：340-343.
[118]王本祥.现代中药药理学[M].天津科学技术出版社,1999,1227-1230.
[1]Yin F., Zhang Y. N., Yang Z. Y. and Hu L. H.. Nine New Dammarane Saponins from Gynostemma pentaphyllum. Chemistry & Biodiversity [J],2006,3:771-782.
[2]Yin F., Hu L. H., Lou F. C. and Pan R. X.. Dammarane-Type Glycosides from Gynostemma pentaphyllum. J. Nat. Prod. [J],2004,67:942-952.
[3]Iwamoto M., Fujioka T., Okabe H., Mihashi K., and Yamauchi T.. Studies on the Actinostemma lobatum MAXIM. I. Structures of Actinistemmosides A, B, C and D. Dammarane Triterpene Glycosides Isolated from the Herb [J]. Chem. Pharm. Bull.1987,35(2):553-561.
[4]沈兆邦,王成章,陈祥,在原重信.一种新型的绞股蓝皂苷[J].林产化学与工业,1993,13(4)：265-269.
[5]Yin F., Hu L. H., Six New Triterpene Saponins with a 21,23-Lactone Skeleton from Gynostemma pentaphyllum. Helvetica Chimica Acta [J].2005,88(5):1126-1134.
[6]Piacente S. and Pizza C., New Dammarane-Type Glycosides from Gynostemma pentaphyllum [J]. J. Nat. Prod.,1995,58(4),512-519.
[7]Wang J. H., Li W., Sha Y., Tezuka Y, Kadota S. and Li X.. Triterpenoid Saponins from Leaves and Stems of Panax quinquefolium L. J. Asian Nat. Prod. Res [J].2001,3,123-130.
[8]郭绪林,王铁军,边宝林.长梗绞股蓝的化学成分研究[J].药学学报,1997,32(7)：524-529.
[9]Takemoto T., Arihara S., Nakajima T., Okuhira M.. Studies on the Constituents of Gynostemma pentaphyllum Makino. I. Structures of Gypenoide I-XIV. Yakugaku Zassh [J].1983,103(2):173-185.
[10]Yoshikawa K., Takemoto T., Arihara S.. Studies on the Constituents of Cucurbitaceae Plants. XV. On the Saponin Constituents of Gynostemma pentaphyllum Makino. (10) [J]. Yakugaku Zassh,1986,106(9): 758-763.
[11]王建忠,王锋鹏.川党参的化学成分研究[J].天然产物研究与开发,1996,8(2)：8-12.
[12]李巍,李铣.文冠果果柄的化学成分研究[J].中草药,2008,38(3)：334-337.
[13]梁龙,钟炽昌,萧淖殷.傣药“亚洛轻”化学成分研究[J].中草药,1999,25(5)：236-238.
[14]孙凯,李铣.南葶苈子的化学成分[J].沈阳药科大学学报,2003,20：419-421.
[1]Lammer C, Wagerer S, Saffrich R, et al. The cdc25B phosphatase is essential for the G2/M phase transition in human cells [J]. J Cell Sci,1998,11(16):2445-2453.
[2]赵斌,葛金芳,朱娟娟.小议在MTT法测细胞增殖抑制率中IC50的计算方法[J].安徽医药,2007,11(9)：834-835.
[3]王光兰,崔俊生.人参皂苷Rg3诱导肺癌细胞NCI-H460凋亡的机制[J].吉林大学学报,2008,34(6)： 980-985.
[4]陈迪,倪劲松.人参皂苷Rg3对乳腺癌细胞MCF-7诱导凋亡作用[J].吉林大学学报,2008,34(4)： 610-614.