Structures of Phytosterols and Triterpenoids with Potential Anti-Cancer Activity in Bran of Black Non-Glutinous Rice

Structures of some bioactive phytochemicals in bran extract of the black rice cv. Riceberry that had demonstrated anti-cancer activity in leukemic cell line were investigated. After saponification with potassium hydroxide, separation of the unsaponified fraction by reversed-phase high performance liquid chromatography (HPLC) resulted in four sub-fractions that had a certain degree of anti-proliferation against a mouse leukemic cell line (WEHI-3 cell), this being IC₅₀ at 24 h ranging between 2.80–467.11 μg/mL. Further purification of the bioactive substances contained in these four sub-fractions was performed by normal-phase HPLC. Structural characterization by gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS) and nuclear magnetic resonance spectroscopy (NMR) resulted in, overall, the structures of seven phytosterols and four triterpenoids. Four phytosterols, 24-methylene-ergosta-5-en-3β-ol, 24-methylene-ergosta-7-en-3β-ol, fucosterol, and gramisterol, along with three triterpenoids, cycloeucalenol, lupenone, and lupeol, were found in the two sub-fractions that showed strong anti-leukemic cell proliferation (IC₅₀ = 2.80 and 32.89 μg/mL). The other sterols and triterpenoids were campesterol, stigmasterol, β-sitosterol and 24-methylenecycloartanol. Together with the data from in vitro biological analysis, we suggest that gramisterol is a significant anti-cancer lead compound in Riceberry bran extract.     

A Study on Isolation of Chemical Constituents from Sophora Flavescens Ait. and Their Anti-Glioma Effects

Background

Sophora flavescens Ait. is a traditional Chinese medicine with a long history in China. It is mainly used in the treatment of heat dysentery and similar ailments in the clinical. The objective of this paper was to isolate, purify and identify alkaloids from Sophora flavescens Ait. and to explore their inhibitory effects on C6 glioma cells.

Materials and Methods

Column chromatography, extraction and NMR spectroscopy were used to structurally identify the isolated compounds. MTT assay and flow cytometry were used to detect the inhibitory effect of matrine on C6 cells.

Results

Three compounds were isolated from Sophora flavescens Ait., namely matrine, oxymatrine and lupeol. Different concentrations of matrine solution all had inhibitory effects on growth of C6 cell lines, which showed apparent dose-effect relationship. Compared with the control group, proportion of G0/G1 phase cells increased in each matrine concentration group to a maximum of 79.8%; proportion of S phase cells reduced, and proportion of G2/M phase cells declined slightly to a minimum of 6.3%, suggesting that after the action of matrine proliferation of C6 cells was significantly inhibited and the cells were arrested in the G1 phase.

Conclusion

We concluded that Sophora flavescens Ait. has an inhibitory effect on C6 cell proliferation.     

RP-HPLC测定枸骨叶中羽扇豆醇的含量

目的:建立RP-HPLC法测定枸骨叶中羽扇豆醇含量的方法,为枸骨叶的质量分析和评价提供科学依据。方法:采用Agilent Zorbax ODS柱(4.6 mm×150 mm,5μm),柱温为25℃,乙腈-水(8∶2)为流动相,在流速1.0mL.m in^-1和紫外检测波长210 nm下进样20μL对枸骨叶甲醇提取物进行HPLC分析。结果:羽扇豆醇在0.25~12.7μg(r=0.999 8)呈良好的线性关系,平均加样回收率为100.2%(RSD 4.3%)。结论:本方法简便,重复性好,为评价枸骨叶药材的质量提供科学依据。     

蛇葡萄根化学成分的研究

从蛇葡萄Ampelopsissinica根中分得6个成分,经光谱分析鉴定为羽扇豆醇,β-谷甾醇,胡萝卜甙,儿茶素,蔗糖,棕榈酸。羽扇豆醇系首次从该属发现。     

Antiangiogenic activity of lupeol from Bombax ceiba

In the search for antiangiogenic agents from medicinal plants used in Vietnam, a methanol extract of the stem barks of Bombax ceiba was found to exhibit a significant antiangiogenic activity on in vitro tube formation of human umbilical venous endothelial cells (HUVEC). Bioactivity-guided fractionation and isolation carried out on this extract afforded lupeol as an active principle. At 50 and 30 microg/mL lupeol showed a marked inhibitory activity on HUVEC tube formation while it did not affect the growth of tumor cell lines such as SK-MEL-2, A549, and B16-F10 melanoma.     

山椒子主要化学成分的研究

目的：了解山椒子的化学成分。方法：分离纯化采用ＳｅｐｈａｄｅｘＬＨ２０凝胶过滤及硅胶柱色谱方法,结构鉴定采用ＨＮＭＲ,^１３ＣＮＭＲ分析及与文献有关数据比较。结果：从山椒子树皮中鉴定出２个三萜类化合物,ｓｕｂｅｒｏｓｏｌ和ｌｕｐｅｏｌ。结论：为首次从该属植物中分得。     

Lupeol及其衍生/修饰物对肝癌细胞增殖影响的研究

目的:探讨lupeol及其结构改造后衍生/修饰物对肝癌细胞HepG2和SMMC7721细胞增殖的作用。方法:对lupeol进行化学结构修饰,然后应用MTT方法检测lupeol及其衍生/修饰物对HepG2和SMMC7721细胞增殖的作用。结果:MTT数据显示,lupeol及其衍生/修饰物均能抑制HepG2和SMMC7721细胞增殖,并且具有浓度依赖性。Lupeol、H1、T1和T2的IC50分别为40μmol/L-50μmol/L、约120μmol/L、约50μmol/L和70μmol/L-80μmol/L。结论:lupeol结构改造后,并没有提高其对肝细胞肝癌细胞增殖的抑制作用。     

多胺对白桦悬浮细胞生长和三萜积累的影响

目的 分析外源多胺对白桦悬浮细胞生长和三萜积累的影响。方法 在白桦悬浮细胞的生长末期添加0.1 mmol/L和1.0 mmol/L的腐胺（Put）、精胺（Spm）和亚精胺（Spd）,采用比色法和RT-PCR方法分析白桦三萜量及其合成关键酶羽扇豆醇合酶（LUS）基因的表达。结果 除1.0 mmol/L Spm处理使白桦悬浮细胞的活力和干质量积累降低外,其他处理均提高了白桦悬浮细胞的活力、干质量积累和三萜产量,且随着处理时间的延长,干质量积累和三萜产量呈增加趋势。其中1.0 mmol/L Put在处理的第2天对细胞干质量和三萜产量的促进效应最大,分别比对照增加了38.89%和116.35%。LUS基因的RT-PCR检测结果进一步证实了多胺对白桦三萜积累的促进作用。结论 Put可有效促进白桦悬浮细胞生长和三萜积累。     

狼毒大戟地上部分化学成分研究

目的 研究新鲜狼毒大戟Euphorbia fischeriana地上部分的化学成分。方法 采用硅胶柱色谱、高效液相及重结晶等方法进行分离纯化,通过波谱分析的方法鉴定化合物结构。结果 从新鲜狼毒大戟地上部分甲醇提取液正己烷萃取物中分离得到14个化合物,分别鉴定为23(E)-25-methoxycycloart-23-en-3β-ol(1)、24-hydroperoxycycloart-25-en-3β-ol(2)、25-hydroperoxycycloart-23-en-3β-ol(3)、羽扇豆醇(4)、24-亚甲基环阿尔廷醇(5)、大戟醇(6)、24-methylenecycloartane-3,28-diol(7)、钝叶甾醇(8)、叶绿醇(9)、jolkinolide A(10)、2,4-二羟基-6-甲氧基-3-甲基苯乙酮(11)、24-亚甲基-9,19-环-3-羊毛甾酮(12)、12-去氧佛波醇-13-十六酸酯(13)、butyrospermol(14)。结论 其中化合物9为首次从该属植物中分离得到,化合物1~3、7、8为首次从该植物中分离得到。     

小娃娃皮的化学成分研究

目的研究川东特有植物小娃娃皮Daphne gracilis全株的化学成分。方法利用正、反相硅胶柱色谱、薄层色谱等方法进行分离纯化,通过多种波谱分析方法鉴定化合物的结构。结果从小娃娃皮全株95%乙醇提取液中分离得到15个化合物,分别鉴定为β-谷甾醇(1)、木栓酮(2)、豆甾-4-烯-3-酮(3)、豆甾-4-烯-3,6-二酮(4)、羽扇豆醇(5)、豆甾醇(6)、1,3-二硬脂酸甘油酯(7)、1-二十二烷酸甘油酯(8)、十八醇(9)、植醇(10)、去氢双松柏醇(11)、epoxyconiferyl alcohol(12)、齐墩果瑞香酮(13)、14-姜酮醇(14)、马台树脂醇(15)。结论 15个化合物均为首次从该植物中分离得到,其中化合物2~4、7~9、11、12、14为首次从该属植物中分离得到;化合物11、12、14为首次从该科植物中分离得到。