超声辅助提取兖州卷柏生物碱及其对H22肝癌细胞增殖抑制影响
Effect of ultrasonic-assisted extraction and anti-proliferation activities on hepatocellular carcinoma cell H22 of alkaloid from Selaginella involvens
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摘要: 目的:研究超声波辅助兖州卷柏生物碱提取工艺,以及兖州卷柏生物碱对小鼠H22肝癌细胞的增殖抑制作用。方法:采用超声辅助乙醇浸提法对兖州卷柏生物碱进行提取,并设计正交实验对提取工艺进行优化。采用MTT法研究兖州卷柏生物碱对小鼠H22肝癌细胞增殖抑制活性。结果:超声辅助兖州卷柏提取最佳工艺为:乙醇浓度70%、固液比1:40、超声时间60min、超声功率400W和超声温度60℃。最高提取率达1.18%。兖州卷柏生物碱对小鼠H22肝癌细胞增殖抑制效果随浓度增大和培养时间增长而增加,其中细胞培养48h,生物碱浓度为8mg/mL时,对肝癌细胞增殖抑制率达53.25%。结论:超声辅助法是兖州卷柏生物碱提取的有效方法,兖州卷柏生物碱对肝癌H22细胞有明显抑制作用。Abstract: Objective:An ultrasonic technology was employed to extract alkaloid from Selaginella involvens (ASI) .Furthermore, anti-proliferation activity on hepatocellular carcinoma cell H22 of mouse in vitro of ASI was observed.Methods:The alkaloids from Selaginella involvens were extracted by using ultrasonic-assisted ethanol.An orthogonal experiment was carried out to optimize the extraction technology.Furthermore, the proliferation inhibiting of hepatocellular carcinoma cell H22 was determined by MTT assay.Results:The optimum extraction conditions of polysaccharides was as follows:ethanol concentrations of 70%, solid-liquid ratio of 1:40, ultrasonic duration of 60min, ultrasonic power of 400W and ultrasonic temperature of 60℃.The highest alkaloid extraction rate reached 1.18%.H22 cell proliferation rate gradually increased with the alkaloid concentration and time increasing.Under the condition of alkaloid condensation of 8mg/mL, the growth inhibition was the highest after 48h and reached 53.25%.Conclusions:Ultrasonic method was a very effective method of extraction alkaloid from Selaginella involvens.The alkaloid could inhibit the growth of hepatocellular carcinoma cell H22 of mouse in vitro.
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