【作者】 张宇；

【导师】 罗擎英； 胡志上；

【作者基本信息】 四川农业大学 ， 农业硕士（专业学位）， 2020， 硕士

【摘要】 花椒（Zanthoxylum bungeanum Maxim）籽是芸香科植物花椒树的果实,是花椒香辛料生产过程中的主要副产物。为对花椒籽的充分开发利用提供依据,本研究首先以汉源花椒籽为原料,通过响应面法优化花椒籽油的提取工艺并对花椒籽油理化性质进行测定。在此基础上,本研究对花椒籽油的体外抗氧化能力进行检测,并建立高脂膳食小鼠模型,观察花椒籽油对高脂膳食小鼠脂质代谢的影响,并对花椒籽油降脂分子机制进行探索。主要研究结果如下:（1）花椒籽油提取工艺优化和理化性质的研究以汉源花椒籽为原料,石油醚（沸程60℃～90℃）为溶剂,采用超声辅助法对花椒籽油进行提取,在单因素实验基础上通过响应面法得到最佳参数:超声功率270 W,料液比1∶16 g/m L,提取时间为75 min。此条件下有最大提取率为22.94%,超声辅助提取法缩短提取时间,降低料液比。使用气相色谱-质谱联用法对花椒籽油脂肪酸成分进行分析,并按照植物油食品安全国标GB/2716-2018对花椒籽油理化性质的测定。实验结果表明,超声辅助提取法显著提高花椒籽油提取效率,汉源花椒籽油中富含不饱和脂肪酸,花椒籽油的酸价较高,可以进一步精炼。（2）花椒籽油生物活性功能的研究通过花椒籽油对DPPH自由基清除能力的测定,对花椒籽油的抗氧化活性进行评价。花椒籽油半抑制浓度IC₅₀为1.87 mg/m L,低于对照组BHT(半抑制浓度IC₅₀为0.31mg/m L),具有一定的抗氧化性。动物试验中,将昆明小鼠随机分组:对照组、模型组、低剂量组（0.1 m L/d）、中剂量组（0.2 m L/d）、高剂量组（0.3 m L/d）和药剂组（辛伐他汀水溶液0.3 m L/d）,通过高脂膳食喂养小鼠建立高血脂小鼠模型。结果表明,高脂模型组小鼠与对照组相比,体重显著上升,血清中胆固醇（TC）、低密度脂蛋白胆固醇（LDL-C）和甘油三酯（TG）含量显著增高,高密度脂蛋白胆固醇（HDL-C）显著降低,模型组小鼠血脂代谢异常。与模型组相比,低剂量血脂各项指标有所下降但效果并不显著;中剂量小鼠体重显著下降,TC和LDL-C含量显著下降,但TG和HDL-C含量变化不显著;而高剂量组小鼠血清中TC、LDL-C和HDL-C都有显著变化。结果表明,花椒籽油对血脂代谢异常具有调节作用,并且摄入量在0.3 m L/d时,效果最好。花椒籽油对小鼠肝脏抗氧化指标测定结果表明,高脂膳食条件下小鼠血脂代谢紊乱可能导致肝脏超氧化物歧化酶（SOD）、过氧化氢酶（CAT）活性降低,同时增加脂质过氧化物丙二醛（MDA）在肝脏中的积累。而不同剂量花椒籽油摄入,均可改善小鼠肝脏抗氧化指标,并且高剂量组小鼠效果最好。此外,通过免疫印迹试验检测小鼠肝脏中胆固醇合成限速酶HMG-Co A还原酶和清道夫受体蛋白CD36表达情况。结果表明,摄入花椒籽油后,小鼠HMG-Co A还原酶表达水平显著下降,而CD36蛋白表达水平上升,这可能是改善脂代谢异常的潜在分子机制之一。更多还原

【Abstract】 In this study,Han Yuan Zanthoxylum bungeanum Maxim seeds were used as raw materials to optimize the extraction process of seed oil by response surface methodology.The physicochemical properties of seed oil were determined,then the antioxidant capacity of Zanthoxylum bungeanum Maxim seed oil was test.Next,a hyperlipemia mice model was established by high-fat diet to observe the effect of Zanthoxylum bungeanum Maxim seed oil on lipid metabolism in high-fat diet mice,and explore the molecular mechanism.The main results are as follows:（1）Study on optimization of extraction and physicochemical properties of Zanthoxylum bungeanum Maxim Seed OilAs raw material,the Zanthoxylum bungeanum Maxim seed oil was extracted by using ultrasound-assisted extracted method.Based on the single factor experiment,the best parameters were obtained by response surface method.The optimized extraction methods were as follow:ultrasonic power 270 W,material-liquid ratio 1:16 g/m L,extraction time 75min,with the maximum extraction rate reached 22.94%.Ultrasound-assisted extraction method greatly shortens the extraction time and reduces the material-liquid ratio.The fatty acid compositions of Zanthoxylum bungeanum Maxim seed oil were analyzed by gas chromatography-mass spectrometry（GC-MS）,and physicochemical properties were determined according to the national standard GB/2716-2018.The results showed that the ultrasound-assisted extracted method significantly improves the efficiency of extraction.Han yuan Zanthoxylum bungeanum Maxim seed oil is rich in unsaturated fatty acids,but the acid value of seed oil is high.（2）Study on the biological activity of Zanthoxylum bungeanum Maxim seed oilThe antioxidant activity of Zanthoxylum bungeanum Maxim seed oil was evaluated by determination of DPPH free radical scavenging ability.The IC₅₀of Zanthoxylum bungeanum Maxim seed oil is 1.87 mg/m L,lower than that of control group BHT(IC₅₀is 0.31 mg/m L),indicating that Zanthoxylum bungeanum Maxim seed oil has certain antioxidant activity.In animal experiments,KM mice were randomly divided into control group,model group,low-dose group（0.1 m L/d）,medium-dose group（0.2 m L/d）,high-dose group（0.3m L/d）and positive control group（0.3 m L/d Simvastatin）.The hyperlipidemia mouse model was established by a high-fat diet.The results showed that,the mice in the model group gained significantly more weight,significantly increased serum cholesterol（TC）,low-density lipoprotein cholesterol（LDL-C）and triglyceride（TG）,and significantly decreased high-density lipoprotein cholesterol（HDL-C）,indicating that the a hyperlipemia mice model was established,with abnormal lipid metabolism.Compared with the model group,the mice in the low-dose group had significantly lower body weight and lower blood lipid,but the effect was not significant.The body weight of medium-dose mice decreased significantly,and the contents of TC and LDL-C decreased significantly,but the contents variation of TG and HDL-C were not significant.The contents of TC,LDL-C and HDL-C in serum were significantly changed in the high-dose group.The results showed that Zanthoxylum bungeanum Maxim seed oil had a regulating effect on abnormal lipid metabolism,and the best effect was achieved when intake was 0.3 m L/d.The results of the determination of antioxidant index of the liver of mice with Zanthoxylum bungeanum Maxim seed oil showed that the dyslipidemia of the liver would reduce the activity of superoxide dismutase（SOD）and catalase（CAT）,and lead to the accumulation of lipid peroxides malondialdehyde（MDA）in the liver.And the intake of Zanthoxylum bungeanum Maxim seed oil at different doses all improved the antioxidant index of mice liver,and the optimum efficiency was achieved when high dose was given.In addition,the expression of HMG-Co A reductase and CD36 protein in mice liver was detected by Western-blot.The results showed that,after ingesting Zanthoxylum bungeanum Maxim seed oil,the expression level of HMG-Co A reductase in mice decreased significantly while the expression level of CD36 protein increased.This may be one of the potential molecular mechanisms of Zanthoxylum bungeanum Maxim seed oil in regulating abnormal lipid metabolism.更多还原