响应面法优化水溶性虎奶菇多糖提取工艺及其对胃癌细胞的抑制作用
Optimization of Extraction of Water-Soluble Polysaccharide from Pleurotus Tuber by Response Surface Method and Their Inhibitory Effects on Gastric Cancer Cells
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摘要: 目的:研究水溶性虎奶菇多糖的提取方法及其对胃癌细胞生长的影响。方法:响应面法优化多糖提取工艺条件,WST-1细胞增殖检测试剂盒以及流式细胞术研究不同浓度水溶性虎奶菇菌核多糖溶液对胃癌细胞增殖率和早期凋亡的影响。结果:虎奶菇多糖的最佳提取条件为料液比1:28 g/mL,提取时间为3.5 h,提取温度98℃,在此条件下多糖得率为2.59%±0.5%。虎奶菇多糖在质量浓度15~500 μg/mL范围内,对胃癌细胞增殖的抑制率最高达30.3%,且成剂量依赖关系。Annexin V-FITC/PI实验结果表明,随着多糖给药剂量的增加,凋亡和坏死细胞不断增加。结论:虎奶菇多糖对体外培养的人SGC-7901胃癌细胞增殖具有抑制作用,并能诱导肿瘤细胞凋亡。Abstract: Objective:To optimize the extraction of polysaccharide from Mushroom Sclerotia of pleurotus tuber regium and to study their inhibitory effects on gastric cancer cells. Methods:Response surface method was used to optimize the extraction process for Mushroom Sclerotia polysaccharide. WST-1 Cell Proliferation and Cytotoxicity Assay Kit and flow cytometry were used to investigate the effects of the polysaccharide with different concentrates on cell proliferation and early apoptosis. Results:The optimal extraction conditions as follows:Material-to-liquid ratio,1:28 g/mL,extraction time,3.5 h,and extraction temperature 98℃. Under the conditions,the maximum yield of polysaccharide was 2.59%±0.5%. The polysaccharide posseses anti-proliferation effect on SCG-7901 in a does-depend manner in a range of 15~500 μg/mL,and the maximum of inhibition rate was 30.3%. Annexin v-fitc/PI results showed that apoptosis and necrotic cells increased with the increase of polysaccharide concentrates. Conclusions:polysaccharide of pleurotus tuber can induce apoptosis of cancer cells.
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