【作者】 商红军；

【导师】 孟宪军；

【作者基本信息】 沈阳农业大学 ， 食品科学， 2011， 博士

【摘要】 五味子含有多种营养成分,是一种具有很大开发前景的药食兼用植物。五味子中的主要活性物质是木脂素,而五味子乙素在五味子木脂素中含量较高,是主要的功能成分。目前研究发现,五味子乙素具有很强的抗氧化性和清除自由基作用,能够预防阿霉素引起的心脏毒性,其脂质体作为药物新型剂,能够提高药物的靶向性,降低药物毒性。本文以辽宁产北五味子为原料,通过正交实验研究了五味子中功能性成分五味子乙素的提取方法,采用大孔树脂法进行纯化,并在此基础上对五味子乙素提取液的清除自由基、抑菌作用进行初步研究；研究了五味子乙素脂质体和五味子乙素—阿霉素脂质体的制备工艺；对制备的五味子乙素脂质体进行了理化性质研究；对五味子乙素—阿霉素脂质体进行了理化性质、药动力学特性进行了研究；对五味子乙素的抗肿瘤作用进行了初步研究。主要研究结果如下：1.溶剂法提取五味子乙素条件为：溶剂浓度90%,液料比7：1,处理时间2.5h,次数3次：超临界CO2提取五味子乙素条件为：压力30 Mpa,温度40℃,粒度40目,时间2.0h；微波辅助提取五味子乙素条件为：微波功率为500 w,液料比为10：1,处理时间为8 min；超声波提取五味子乙素条件为：超声波功率180W,液料比为10：1,处理时间为40min。2.实验中所用的树脂中,AB-8树脂的吸附和解吸能力都最强,适宜于五味子乙素的分离纯化。用AB-8树脂分离纯化五味子提取物时,其工艺参数为：上样浓度0.09mgmL-上样流速为1.0 mL.min-1,静态吸附时间为150mmin,解吸流速为0.5 mL.min-1。3.采用旋转蒸发薄膜法制备五味子乙素脂质体。通过单因素及正交试验,确定最佳制备工艺为：大豆磷脂与胆固醇质量比为1：0.1,五味子乙素载量1%,旋转蒸发温度为65℃,pH值为6.9。在此条件下,五味子乙素脂质体的粒度分布为70～200 nm,平均粒径值为(132.5士19.6)nm(n=3),Zeta电位为(-37.3±2.5)mV,包封率为92.4%。4.采用旋转蒸发薄膜法制备五味子乙素—阿霉素脂质体。取五味子乙素为0.2g时,通过单因素及其正交试验确定最佳制备工艺为：大豆磷脂与胆固醇的比为1：0.05,大豆磷脂与阿霉素的比为2：0.01,旋转蒸发温度为60℃,pH值为7.4。在此条件下制备的五味子乙素-阿霉素脂质体粒度小,包封率高。粒度分布为50～200 nm,平均粒径值为(135.2±19.6)nm(n=3),Zeta电位为(-38.6±2.5)mV,包封率为96.45%。五味子乙素-阿霉素脂质体的药动力学试验结果表明,五味子乙素-阿霉素脂质体能够提高阿霉素的血浆药物浓度,延长药物在血液的停留时间,以获得更充足的时间到达靶向部位,提高治疗效果。5.急性毒性试验结果：五味子乙素-阿霉素脂质体LD50=52.8mg/kg,阿霉素脂质体LD50=41.5mg/kg,阿霉素LD50=18.2mg/kg。由此可初步得知,五味子乙素—阿霉素脂质体能够降低阿霉素的毒性。采用动物S-180实体肿瘤模型,采用尾静脉注射给药法,对五味子乙素的抗肿瘤作用进行了初步研究。结果表明,本试验制备的五味子乙素脂质体抗肿瘤作用并不显著,制备的五味子乙素—阿霉素脂质体的抗肿瘤效果比较显著。6.五味子乙素提取液对自由基的清除效果明显,其中对.OH清除作用大于相同浓度的Vc,其半数清除浓度为0.2 mg.mL-1；对O2·-清除作用小于Vc,其半数清除浓度为0.24 mg.mL-1。五味子乙素提取液对金黄色葡萄球菌、白色念珠菌、沙门氏菌、大肠杆菌、枯草芽孢杆菌均有一定的抑制作用,其中对白色念珠菌和金黄色葡萄球菌的抑制作用最强,最低抑菌浓度为0.25 mg.mL-1,对根霉、黑曲霉、南阳酵母无明显抑菌活性。更多还原

【Abstract】 Schisandra chinensis(Turcz.)Baill has variety of nutrients. It is a good development prospects plant with the use of drug and eating. Lignan which is the main active ingredient of Schisandra chinensis has a high content ofγ-schizandrin. The main functional component isγ-schizandrin. Modern research has shown thatγ-schizandrin has cancer-fighting doxorubicin and scavenging free radicals role. The principal active ingredient are lignan and polysacch-aride. Liposomes as a drug of new agents that can improve drug targeting and reduce drug toxicity.This subject took Schisandra chinensis which was collected in Liao Ning province as material, studied the extract method ofγ-schizandrin by using orthogonal test design, macroporous resin were employed forγ-schizandrin separation and purification. The antioxidant and antimicrobial activities of the extract were also investigated. This study investigated the preparation of theγ-schizandrin liposomes and theγ-schizandrin-doxorubicin liposomes. Research the Physical and Chemical Properties ofγ-schizandrin liposomes, the Physical and Chemical Properties ofγ-schizandrin-doxorubicin liposomes, the drug kinetics. Preliminary study the anti-tumor effect ofγ-schizandrin. The main results were shown as follows:1.The optimum technological conditions of solvent extraction:solvent strength 90%, Liquid:Materal 7:1, processing time 2.5h, extract 3 times; The optimum technological conditions of supercritical CO2 extraction:pressure 30Mpa, temperature 40℃, granularity 40 mesh, time 2h; The optimum extraction conditions of microwave assisted extraction were as follows:power 500 W, solvent-material ratio 10:1, extraction time 8min; The optimum extraction conditions of ultrasonic waves:power180 W, solvent-material ratio 10:1, extraction time 40min.2.AB-8 resin is suitable forγ-schizandrin separation and purification with better capabilities of absorption and desorption in the research. The technological parameters of AB-8 edulcorateγ-schizandrin:concentration ofγ-schizandrin 0.09mg·mL-1; absorption flow rate 1mL·min-1; absorption time 150min; elution rate of 0.5 mL·min-1.3.Adopts rotating evaporation film for offering legal liposomes. Edible with encasulation efficiency as evaluation indexes. By single factor and orthogonal test, for optimum technological conditions 1:0.1,γ-schizandrin grain cindoruk 1%, Rotating evaporation temperature for 60℃, pH 6.9, encapsulation rates 92.4%.4. Prepared by rotating evaporation film legalγ-schizandrin -doxorubicin liposomes. The best preparation process for:Soybean lecithin and cholesterol than for the 1:0.05, Soybean lecithin and than for the adriamycin 2:0.01. In rotating evaporation temperature for 60℃conditions and pH= 7.4. This method of the preparation of fructusγ-schizandrin element of liposomes doxorubicin particle distribution of 50～200 nm. Average particle size value is (135.2±19.6) nm (n=3). Measured by flocculation separation fructusγ-schizandrin doxorubicin liposomes encapsulation efficiency can reach(96.45±1.86)%, Zeta Potential for(-38.6±2.5) mV. Results show that fructusγ-schizandrin doxorubicin can significantly improve the liposome drug concentration of plasma adriamycin. Fructus schisandrae meal can enhance the party of adriamycin, efficacy, together they can prolong drug use in the retention period of blood, to get plenty of time to get to target sites, increases healing done.5. For acute toxicity tests, fructus schisandrae b grain doxorubicin liposomes LD50=52.8 mg/kg, Doxorubicin liposomes LD50= 41.5 mg/kg, Doxorubicin LD50= 1.82 mg/kg. We can see that, Fructusγ-schizandrin to significantly reduce the toxicity of adriamycin. Using animal S-180 solid tumors model, adopt tail intravenous drug laws fructusγ-schizandrin doxorubicin liposomes anti-tumor effect. Results show that, Meat and edible bliposome fructusγ-schizandrin element has certain antitumor function but not significant, doxorubicin liposomes compared with adriamycin, Cancer-fighting improve also was not significant. But fructusγ-schizandrin doxorubicin anti-cancer effects of liposomes significantly higher than doxorubicin.6.γ-schizandrin extract had effective scavenging scavenging effect of.OH more than the same concentration of Vc, the IC50 is 0.2 mg.mL-1, scavenging effect of O2.- less than Vc, the IC50 is 0.24 mg.mL-1.γ-schizandrin extract against Staphylococcus aureus, Candida albicans, Salmonella, E. coli, Bacillus subtilis has a certain extent, of which Candida albicans and Staphylococcus aureus in the strongest inhibition, minimum inhibitory concentration of 0.25 mg.mL-1; on Rhizopus, Aspergillus niger, Nanyang yeast had no significant antibacterial activity.更多还原