东北黑蜂蜂胶黄酮提取工艺优化及抗肿瘤活性研究

发布时间：2018-08-03 16:22

【摘要】：目的:蜂胶是一种蜜蜂生产的被广泛使用天然产品,具有消炎、杀菌、抑制肿瘤等作用为人类的健康做出很大贡献。近年来癌症已成为危害人体健康的首要疾病,目前其主要治疗途径是放疗,但放疗副作用很大,能杀死一部分正常的细胞。蜂胶作为一种具有抗肿瘤生物学活性的天然产物,可诱使癌细胞凋亡对正常细胞没有影响。本课题通过正交试验探究蜂胶醇提物黄酮类化合物的优化方案;观察东北黑蜂蜂胶醇提取物(EEP)对荷瘤小鼠免疫活性的影响和肿瘤抑制率,探讨EEP的抗肿瘤活性。方法:制备H22模型小鼠,将蜂胶用乙醇提取做正交试验优化实验方案再对东北黑蜂蜂胶进行提取,进行体内实验。碳廓清试验和抗肿瘤实验,把小鼠分成阴性对照组、蜂胶高剂量组组、蜂胶中剂量组、蜂胶低剂量组、环磷酰胺组五组,连续给药7天,称体重。注射印度墨汁,测定碳廓清的吞噬指数。最后一次给药后脱颈处死解剖取脏器,再精确称量其重,计算黑蜂蜂胶提取物肿瘤抑制率。结果:乙醇提取东北黑蜂蜂胶黄酮化合物的优化方案是温度为常温,固液比为1:10,乙醇浓度为90%,时间为12 h;将碳廓清试验中的黑蜂蜂胶剂量组的吞噬指数与阴性对照组做比较,具有统计学意义(P0.05);抗肿瘤实验中的黑蜂蜂胶的三个组的瘤重分别与阴性对照组作比较,有统计学意义。结论:优化后的方案使黄酮提取成本降低,符合实际需要;碳廓清实验说明东北黑蜂蜂胶醇提取物能够提高H22小鼠的免疫活性;抗肿瘤实验说明东北黑蜂蜂胶醇提取物对实体瘤的生长在一定程度上有很强的抑制作用。综上实验表明东北黑蜂蜂胶的乙醇提取物黄酮类化合物抗肿瘤活性显著。
[Abstract]:Objective: propolis is a widely used natural product produced by honeybees. It has the functions of anti-inflammatory, bactericidal and tumor inhibition, and makes a great contribution to human health. In recent years, cancer has become the primary disease that endangers human health. At present, radiotherapy is the main treatment way, but the side effects of radiotherapy are very big, which can kill some normal cells. Propolis, as a natural product with anti-tumor biological activity, can induce cancer cell apoptosis to have no effect on normal cells. In this paper, the optimum scheme of flavonoids extracted from propolis alcohol was studied by orthogonal experiment, and the effect of (EEP) on the immune activity and tumor inhibition rate of mice bearing tumor was observed, and the anti-tumor activity of EEP was discussed. Methods: H22 model mice were prepared. The ethanol extraction of propolis was optimized by orthogonal experiment. The mice were divided into negative control group, high dose propolis group, middle dose propolis group, low dose propolis group and cyclophosphamide group. The phagocytic index of carbon clearance was measured by injection of Indian ink. After the last administration, the organs were dissected and weighed accurately, and the tumor inhibition rate of the extract of Propolis was calculated. Results: the optimum scheme of ethanol extraction was as follows: the temperature was normal temperature, the ratio of solid to liquid was 1: 10, the concentration of ethanol was 90 and the time was 12 hours. The phagocytic index of the dose group of propolis in carbon clearance test was compared with that of the negative control group. There was statistical significance (P0.05); the tumor weight of the three groups of black bee propolis in the anti-tumor experiment was compared with the negative control group, which had statistical significance. Conclusion: the optimized scheme can reduce the cost of flavonoids extraction and meet the practical needs, and the carbon clearance experiment shows that the alcohol extract of Propolis can improve the immune activity of H22 mice. The antitumor experiment showed that the alcohol extract of Propolis had a strong inhibitory effect on the growth of solid tumor to a certain extent. The experimental results showed that the flavonoids from the ethanol extract of Propolis were significant in anti-tumor activity.
【学位授予单位】：佳木斯大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TQ464;R965

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