石榴皮总黄酮的提取及提取物对小鼠免疫功能影响的实验研究
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摘要
目的:探索研究乙醇提取石榴皮中总黄酮的最佳工艺条件,探讨石榴皮总黄酮对小鼠免疫功能的影响。方法:1)采用正交实验设计,以芦丁为标准品,用分光光度法对新疆石榴皮中的总黄酮含量进行测定,并研究其最佳提取工艺条件;2)应用环磷酰胺复制免疫低下小鼠模型,同时以石榴皮总黄酮提取物作为受试物灌胃10天,研究石榴皮总黄酮对小鼠的外周血白细胞数、脾脏及胸腺指数、抗体分泌细胞功能、血清溶血素水平、巨噬细胞吞噬功能、脾脏淋巴细胞转化功能等免疫指标的影响。结果:1)石榴皮中总黄酮含量为67.737-100.234 mg/g,平均为81.737mg/g。乙醇提取石榴皮总黄酮的最佳工艺条件是:过40目筛网的石榴皮粉用70%乙醇在80℃下回流提取3h,其中主要因素为原料粒度的大小,其次为提取温度和乙醇浓度,影响最小的因素为提取时间;2)石榴皮总黄酮粗提物的三个剂量组及石榴皮总黄酮精提物组均能提高免疫力低下小鼠的外周血白细胞数。石榴皮总黄酮粗提物高剂量组能提高免疫力低下小鼠的脾脏指数、抗体分泌细胞功能、血清溶血素水平以及脾淋巴细胞转化功能。石榴皮总黄酮粗提物的中、高剂量组及石榴皮总黄酮精提物组均能提高免疫力低下小鼠的巨噬细胞吞噬功能。结论:1)原料粒度、提取温度及乙醇浓度对石榴皮总黄酮溶出影响较大,应加以控制;2)石榴皮总黄酮粗提物对免疫力低下小鼠的免疫功能具有一定的增强作用。
Objective:To study the best conditions of extraction technology for total flavonoids from pomegranate peel with ethanol,and to study the immunomodulation of total flavonoids of pomegranate peel in immunosuppressed mice.Methods:1)Regarded rutin as the contrast calibre,determined content of total flavonoids in pomegranate peel by spectrophotometry,and the best conditions of extraction technology were optimized with orthogonal design.2)The immunosuppressed mice induced by Cyclophosphamide were administrated with total flavonoids extraction of pomegranate peel for 10 days.The white blood cell(WBC)of peripheral blood,the immune organ weights,the ability of antibody secreting cell,the level of serum hemolysin,the phagocytosis ability of Macrophage and the ability of lymphocyte transformation were measured.Results:1)The total flavonoids of pomegranate peel contained 67.737mg/g-100.234 mg/g,average was 81.737mg/g.The optimum extracting conditions was pomegranate peel powder getting 40 mesh size,adding 70%alcohol,refluxing and extracting for 3h at 80℃.The main factor was particle size of material,then temperature of extraction and concentration of alcohol, the minimal factor was time of extraction.2)The WBC of immunosuppressed mice were increased by three groups of crude extraction of total flavonoids and the group of refined extraction of total flavonoids of pomegranate peel.The immune organ weights,the ability of antibody secreting cell,the level of serum hemolysin and the lymphocyte transformation were strenghened at high does of crude extraction of total flavonoids.The phagocytosis ability of Macrophage were improved at middle and high does groups of crude extraction of total flavonoids and the group of refined extraction of total flavonoids of pomegranate peel.Conclusions:I)Particle size of material,temperature of extraction and concentration of alcohol should be strictly controlled because of their large influences on the dissolution of total flavonoids.2)The crude extraction of total flavonoids of pomegranate peel can improve the immune function in immunosuppressed mice induced by Cyclophosphamide.
Objective:To study the best conditions of extraction technology for total flavonoids from pomegranate peel with ethanol,and to study the immunomodulation of total flavonoids of pomegranate peel in immunosuppressed mice.Methods:1)Regarded rutin as the contrast calibre,determined content of total flavonoids in pomegranate peel by spectrophotometry,and the best conditions of extraction technology were optimized with orthogonal design.2)The immunosuppressed mice induced by Cyclophosphamide were administrated with total flavonoids extraction of pomegranate peel for 10 days.The white blood cell(WBC)of peripheral blood,the immune organ weights,the ability of antibody secreting cell,the level of serum hemolysin,the phagocytosis ability of Macrophage and the ability of lymphocyte transformation were measured.Results:1)The total flavonoids of pomegranate peel contained 67.737mg/g-100.234 mg/g,average was 81.737mg/g.The optimum extracting conditions was pomegranate peel powder getting 40 mesh size,adding 70%alcohol,refluxing and extracting for 3h at 80℃.The main factor was particle size of material,then temperature of extraction and concentration of alcohol, the minimal factor was time of extraction.2)The WBC of immunosuppressed mice were increased by three groups of crude extraction of total flavonoids and the group of refined extraction of total flavonoids of pomegranate peel.The immune organ weights,the ability of antibody secreting cell,the level of serum hemolysin and the lymphocyte transformation were strenghened at high does of crude extraction of total flavonoids.The phagocytosis ability of Macrophage were improved at middle and high does groups of crude extraction of total flavonoids and the group of refined extraction of total flavonoids of pomegranate peel.Conclusions:I)Particle size of material,temperature of extraction and concentration of alcohol should be strictly controlled because of their large influences on the dissolution of total flavonoids.2)The crude extraction of total flavonoids of pomegranate peel can improve the immune function in immunosuppressed mice induced by Cyclophosphamide.
引文
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