Study on Anti-tumor Effect of Ethanol Extract of Smilax trinervula and Its Different Polar Extract Parts / 中国药房

OBJECTIVE： To study the anti-tumor effect of ethanol extract of Smilax trinervula and its different polar extract parts， and to provide reference for the screening of anti-tumor active parts. METHODS： MTT method was used to detect the inhibitory rates of different concentrations of ethanol extract of S. trinervula， its petroleum ether， ethyl acetate， n-butanol and water layer extract parts to the proliferation of human hepatocellular carcinoma cell line HepG2， human lung cancer cell line A549， human breast cancer cell line MCF-7 and MDA-MB-231， human cervical cancer cell line HeLa and human ovarian cancer cell line HO-8910. Semi-inhibitory concentration （IC50） was calculated. A total of 80 KM mice were subcutaneously inoculated with S180 cell suspension in right forelimb axilla to induce tumor-bearing mice model. The mice were randomly divided into 8 groups： e.g. model group （normal saline， twice a day， i.g.）， cyclophosphamide （positive drug） group （0.025 g/kg， once a day， i.p.）， S. trinervula ethanol extract high-dose， medium-dose and low-dose groups， ethyl acetate part of ethanol extract high-dose， medium-dose and low-dose groups （0.1， 0.05， 0.025 g/kg by extract， twice a day， i.g.）， with 10 rats in each group. The mice in each group were given medicine for consecutive 14 days. The inhibition rate， spleen index and thymus index of mice in each group were measured after fasting for 12 hours after the last administration. RESULTS： In vitro experiments showed that S. trinervula and different polar extracts inhibited the proliferation of 6 kinds of tumor cells in significant dose-effect manner， especially ethyl acetate part of ethanol extract. IC50 of it to tumor cells was 40-210 μg/mL， that of it to MCF-7， MDA-MB-231 and HeLa cells were 70.56， 83.58， 44.67 μg/mL， respectively. In vivo experiments showed that the tumor mass of mice were decreased significantly in each administrations （P＜0.05 or P＜0.01）， the tumor mass of mice in S. trinervula ethanol extract high-dose group， ethyl acetate part of ethanol extract high-dose and medium dose groups were significantly lower than cyclophosphamide group （P＜0.01）. The spleen index of mice was decreased significantly in cyclophosphamide group， S. trinervula ethanol extract high-dose group， ethyl acetate part of ethanol extract high-dose and medium dose groups， compared with model group （P＜0.01）； thymus index of mice in cyclophosphamide group was increased significantly， compared with model group and other administration groups （P＜0.01）. CONCLUSIONS： The ethyl acetate part of ethanol extract of S. trinervula has the best anti-tumor effect and less immunosuppressive effect.

[Studies on chemical constituents of rhizomes of Smilax trinervula].

The chemical constituents were separated and purified from the 70% ethanol extract of Smilax trinervulaby various chromatographic methods including silica gel, Sephadex LH-20, MCI and preparative HPLC. Their structures were obtained and identified by analysis of the spectroscopic data. Compounds 1-11 were separated from this genus for the first time. Compound 12 was obtained from S. trinervula for the first time.

Studies on chemical constituents of rhizomes of Smilax trinervula / 中国中药杂志

The chemical constituents were separated and purified from the 70% ethanol extract of Smilax trinervulaby various chromatographic methods including silica gel, Sephadex LH-20, MCI and preparative HPLC. Their structures were obtained and identified by analysis of the spectroscopic data. Compounds 1-11 were separated from this genus for the first time. Compound 12 was obtained from S. trinervula for the first time.

Phenylpropanoids and neolignans from Smilax trinervula.

A new phenylpropanoid glucoside and two new neolignans, namely (1S, 2R)-1-(3, 4, 5-trimethoxyphenyl)-3-(ß-d-glucopyranosyloxy)-1, 2, 3-propanetriol (1), and (7R, 8R)-4, 7, 9, 9'-tetrahydroxy-3, 5, 3', 5'-tetramethoxy-8-4'-oxyneolignan 4-O-ß-d-glucopyranoside (2) and 3', 9, 9'-trihydroxy-3, 5-dimethoxy-8-O-4'-neolignan-4-O-ß-d-glucopyranoside (3), together with a new natural product (1S, 2R)-1-(3, 4, 5-trimethoxyphenyl)-1, 2, 3-propanetriol (4) and four known compounds (5-8) were isolated from the rhizomes of Smilax trinervula. Their structures were established mainly on the basis of 1D and 2D NMR spectral data, ESI-MS and literature comparisons. Compounds 1-8 were tested in vitro for their cytotoxic activities against 5 human tumor cell lines (SH-SY5Y, SGC-7901, HCT-116, Lovo and Vero). Compounds 7 and 8 exhibited cytotoxic activity against Lovo, with IC50 values of 18.7µM and 16.8µM, respectively.

Structural characterization of steroidal saponins from Smilax trinervula using ultra high-performance liquid chromatography coupled with LTQ-Orbitrap mass spectrometry.

Steroid saponins are a class of naturally existing substances widely distributed in the plants of Smilacaceae. Their biological activities have been attracting the interest of scientists in the chemical field for the past few years. To our best knowledge, there has been no study on structural characterization of steroidal saponins from Smilax trinervula (S. trinervula) using LTQ-Orbitrap mass spectrometry, which could provide an excellent approach for rapid screening of steroidal saponins in other plants of Smilacaceae. An ultra high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC/HRMS) method was therefore developed to characterize steroidal saponins in S. trinervula. This method was operated in both negative and positive ion modes with HRMS, as a result, a total of twenty-two steroidal saponins with three aglycone skeletons were elucidated in the crude extract from the root of S. trinervula. The characteristic-fragment ions could well identify and differentiate the three types of aglycone skeletons (diosgenyl saponins, furostanol saponins, C27-hydroxy diosgenyl saponins).