Effect of soyasapogenol a and soyasapogenol B concentrated extracts on Hep-G2 cell proliferation and apoptosis
Effect of soyasapogenol a and soyasapogenol B concentrated extracts on Hep-G2 cell proliferation and apoptosis
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DOI：
10.1021/jf0731550
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发表时间：
2008-04-23
影响因子：
6.1
通讯作者：
Popovich, David G.
中科院分区：
文献类型：
--
作者：
Zhang, Wei;Popovich, David G.
从大豆粉中提取大豆皂苷（Glycine max (L.)），水解形成大豆皂苷A和B，对人肝癌细胞株Hep-G2的细胞活性进行了抑制生长和诱导凋亡的实验。经HPLC和ESI-MS检测，浓缩大豆皂苷醇A （SG-A）和大豆皂苷醇B （SG-B）提取物分别含有约69.3%和46.2%的苷元（大豆皂苷醇），而粗甲醇提取的大豆皂苷提取物（TS）不含任何可检测量的SG-A或SG-B。MTT活性试验表明，3种提取物对Hep-G2增殖均有一定的影响，其72 h LC50值分别为：TS 0.594 +/- 0.021 mg/mL, SG-A 0.052 +/- 0.011 mg/mL, SG-B 0.128 +/- 0.005 mg/mL。采用流式细胞术、细胞周期分析和共聚焦激光扫描显微镜（CLSM）检测凋亡细胞。细胞周期分析表明，与SG-B相比，SG-A提取物在24 h（25.63 +/- 2.1%）和72 h（47.1 +/- 3.5%）时凋亡细胞的亚g1积聚显著（P < 0.05）增加，而TS提取物仅产生少量亚g1细胞积聚。CLSM证实，在不同LC50浓度下，24 h后所有处理的细胞形态都发生了变化。这些结果表明，与不含大豆皂苷醇的总大豆皂苷提取物相比，主要含有大豆皂苷醇A和B的样品对培养的Hep-G2的增殖抑制能力更强。
The growth inhibition and the induction of apoptosis brought about by soyasaponins extracted from soy flour (Glycine max (L.)) and concentrated for soyasapogenols A and B formed by hydrolysis were tested for cytoactivity in the human hepatocellular carcinoma cell line Hep-G2. Concentrated soyasapogenol A (SG-A) and soyasapogenol B (SG-B) extracts contained approximately 69.3% and 46.2% of their respective aglycones (soyasapogenols) assessed by HPLC and ESI-MS, while the soyasaponin extract (TS), derived from crude methanol extraction, did not contain any detectable amounts of SG-A or SG-B. An MTT viability assay showed that all three extracts had an effect on Hep-G2 proliferation in a dose-response manner with 72 h LC50 values of 0.594 +/- 0.021 mg/mL for TS, 0.052 +/- 0.011 mg/mL for SG-A, and 0.128 +/- 0.005 mg/mL for SG-B. Apoptotic cells were determined by flow cytometry cell cycle analysis and confocal laser scanning microscopy (CLSM). Cell cycle analysis indicated a significant (P < 0.05) greater sub-G1 buildup of apoptotic cells at 24 h (25.63 +/- 2.1%) and 72 h (47.1 +/- 3.5%) for the SG-A extract compared to SG-B, whereas the TS extract produced only a minor buildup of sub-G1 cells. CLSM confirmed a morphological change of all treatments after 24 h, at the respective LC50 concentrations. These results show that the samples that contained mainly soyasapogenols A and B showed a greater ability to inhibit proliferation of cultured Hep-G2 when compared to a total soyasaponin extract that did not contain any soyasapogenols.