文献类型：学位论文

中文题名：马氏珠母贝肉醒酒作用及其醒酒机理的研究

英文题名：Study on Anti-alcoholism Effect and It’s Mechanism of Shellfish Meat from Pinctada Martensii

作者：韩丽娜[1];

机构：[1]广东海洋大学;

导师：秦小明;广东海洋大学

授予学位：硕士

语种：中文

中文关键词：马氏珠母贝;蛋白;糖原;牛磺酸;游离氨基酸;醒酒

外文关键词：Pinctada martensii; Protein; Glycogen; Tautine; Free amino acid; Anti-alcoholis

中文摘要：马氏珠母贝是我国南方海水珍珠养殖的主要贝类,在广东、广西、海南、和台湾沿海等地均有分布。马氏珠母贝肉含有丰富的蛋白质、糖原、必需氨基酸、多种维生素、锌、硒等具有特殊生理作用的矿物质,以及牛磺酸、活性肽等生理活性物质。具有抗肿瘤、抗疲劳、抗衰老等生理作用。另外本课题组的研究结果显示马氏珠母贝肉酶解液的醒酒效果优于牛磺酸对照,推测贝肉中的多肽可能具有醒酒作用,但是其醒酒因子及醒酒机理尚未清楚,也未见有相关报道。本研究在对马氏珠母贝肉粗提物的醒酒作用进行初探的基础上,分析了马氏珠母贝肉糖原、蛋白以及酶解液等组分的醒酒作用,并初步分析了醒酒功效因子的醒酒机理,为马氏珠母贝肉醒酒功能的研究提供理论依据。 试验方法和结果: 1、在系统分析马氏珠母贝肉主要成分的基础上对其醒酒作用进行了初探。其中,蛋白、脂肪、糖原和牛磺酸的含量分别为10.85/%、1.88/%、1.35/%和0.60/%。试验结果表明,与生理盐水组相比,马氏珠母贝肉具有显著的醒酒效果,将小鼠血清中乙醇浓度极显著降低了48.16/%/(P<0.01/)。 2、通过水煮方式将马氏珠母贝肉分为蒸煮液组分/(含牛磺酸4.70mg//ml,糖原8.88mg//ml/)和水不溶性物组分/(含牛磺酸0.01mg//ml,糖原0.31mg//ml/)。采用酶解方法/(中性蛋白酶和木瓜蛋白酶/)对以上水不溶物组分进行酶解,制得酶解液。对马氏珠母贝蒸煮液、水不溶物组分及其酶解液进行醒酒效果分析。试验结果表明,与生理盐水组相比,三者分别将小鼠血清中乙醇浓度显著降低了27.70/%、28.21/%和44.04/%,且水不溶物酶解前后醒酒效果差异显著/(P<0.01/)。经检测马氏珠母贝蒸煮液中亮氨酸、丙氨酸、缬氨酸含量分别为2.27/%、13.41/%和3.71/%。 3、采用三氯甲烷//甲醇混合溶剂/(1:1/)环流方法将马氏珠母贝肉完全脱脂,并通过糖化酶和淀粉酶将糖原充分降解后得到马氏珠母贝纯蛋白/(蛋白99/%/)。采用酶解方法/(中性蛋白酶和木瓜蛋白酶/)对纯蛋白组分进行酶解,制得酶解液。对马氏珠母贝纯蛋白及其酶解液进行醒酒效果分析。试验结果显示,与生理盐水组相比,马氏珠母贝纯蛋白及其酶解液无明显醒酒效果。 4、通过酶解方法/(中性蛋白酶和木瓜蛋白酶/)对马氏珠母贝肉进行酶解,制得酶解液经α-淀粉酶和糖化酶酶解,将糖原充分降解后,再经Sephadex G凝胶柱分离纯化,得到不同分子量的各组分,分别进行小鼠醒酒试验。试验结果显示,与生理盐水组相比,TP2组分及其牛磺酸对照组具有显著的醒酒效果,分别将小鼠血液中乙醇浓度降低15.69/%和27.17/%,其二者间无显著差异,其他组分均无醒酒功效。 5、采用碱液加热方法/(2.3 mol//L KOH,沸水浴加热1h/)制得马氏珠母贝肉糖原/(糖原含量99/%/),其进行醒酒效果分析。试验结果表明马氏珠母贝肉糖原具有明显的醒酒作用,与生理盐水组相比,将小鼠血醇浓度显著降低了56.63/% /(P<0.01/)。 6、通过急性毒性试验、防醉、解酒试验,对马氏珠母贝肉糖原进行了毒理和醒酒作用分析,并分析了其醒酒作用的量效关系。试验结果表明,灌胃剂量为64mg//kg体重时,马氏珠母贝肉糖原无任何毒副作用。马氏珠母贝肉糖原可显著延长小鼠耐受时间,缩短醉酒维持时间。醒酒量效关系分析结果表明,马氏珠母贝肉糖原的最佳醒酒剂量分别为0.80mg//ml,与生理盐水组相比,小鼠血清乙醇浓度显著降低了56.63/%/(P<0.01/)。 7、以乙醇脱氢酶/(ADH/)和乙醛脱氢酶/(ALDH/)为指标,对马氏珠母贝肉糖原醒酒机理进行了初步研究。试验结果显示,相对于对照组,糖原可显著提高ADH和ALDH的酶活力,其中,糖原在所考察的时间点内分别将ADH和ALDH酶活力平均提高了85.46/%和99.84/%。表明马氏珠母贝肉糖原是通过促进ADH和ALDH的酶活,将乙醇和乙醛快速转化为乙酸并排出体外从而达到醒酒作用。

外文摘要：Pinctada martensii is one of the primary seashells of marine pearl cultivation in the southern china. It is fruitful and widely distributed in Guangdong, Guangxi, Hainan, Taiwan and so on. Pinctada martensii has abundant proteins, glycogen, essential amino-acid, varied vitamin, the minerals with particularities effects in physiology, such as zinc and selenium,and some physiological active substance, for example tautine, bioactive peptides etc. Pinctada martensii has many physiological function. These physiological effects include anti-tumor, anti-fatigue, anti-aging and so on. In addition, our research showed that compared with the taurine saline group, the hydrolyzate of Pinctada martensii meat has significant anti-alcoholism effect /(P<0.01/). We spectulated that peptide may have anti-alcoholism effect. However it’s functional factors and mechanism did’t clear, there is no related reports. In this study, based on the preliminary study of sober up role of Pinctada martensii meat extracts, we analyzed the anti-alcoholism effect of the glycogen, protein and its protease hydrolysates, and then we gave the mechanism of anti-alcoholism factor an preliminary analysis, which will provide a scientific theory for the study of the anti-alcoholism effect of Pinctada martensii. Test Method and Results: 1. Based on systematic analysis of the major components of Pinctada martensii meat, we studied its’effect on anti-alcoholism. It has protein 10.85/%,fat 1.88/%,glycogen 1.35/% and taurine 0.60/%. The results show that, compared with the saline group, the meat of Pinctada martensii has significant anti-alcoholism effect /(P<0.01/), the ethanol concentration of blood serum in mice was decreased by 48.16/%/(P <0.01/). 2.The meat of Pinctada martensii was distributed into two parts by boiling, that is cooking liquid components /(taurine 4.70mg//ml, glycogen 8.88mg//ml/) and water-insoluble components /(taurine 0.01/%,glycogen 0.31/%/). By using the neutral protease and papain, we enzymied the water-insoluble components,and got enzymatic hydrolysate. Then we studied their effect on anti-alcoholism. The results showed that, compared with the saline group, all of them have significant anti-alcoholism effect /(P<0.01/), and reduced concentration of ethanol in serum by 27.70/% ,28.21/% and 44.04/%, respectively /(P <0.01/). Separately, there is significant difference between water insoluable residue and its protease hydrolysates. The study of the free amino acids show that ,the cooking liquid of Pinctada martensii has leucine 2.27/%, alanine 13.41/% and valine 3.71/%。 3. By using chloroform // methanol mixed solvent /(1:1/) circulation method, we made the Pinctada martensii meat completely defatted, and then fullly hydrolyzed the glycogen by glucoamylase, and finally gave the purified protein, /(protein 99/%/). In addition, by using the neutral protease and papain, we enzymized the purified protein and got enzymatic hydrolysate. Than we studied their effect on anti-alcoholism. The results show that: the purified protein and its protease hydrolysates did not appear any different comparing with the control. 4. By using the neutral protease and papain, we enzymized the meat of Pinctada martensii and got enzymatic hydrolysate. And then fullly hydrolyzed the glycogen by glucoamylase andα-amylase. It was purified on Sephadex G . The difference molecular wieght components were gained. And we studied their effect on anti-alcoholism. The results showed that, compared with the saline group, TP2 and have significant anti-alcoholism effect /(P<0.01/), and reduced concentration of ethanol in serum by 15.69/% and 27.17/%, respectively /(P <0.01/). Separately, there is no significant difference between them. However, other components did not appear any different comparing with the control. 5. By using hot-alkali extraction method /(2.3 mol //L KOH, boiling water bath heating 1h/), we got the Pinctada martensii glycogen /(glycogen 99/%/). The effect of glycogen commodity on anti-alcoholism was analyzed. The results show that Pinctada martensii glycogen have significant sober up effect /(P<0.01/). Compared with the saline group, the two drugs significantly reduced the ethanol concentration by 56.63/%/(P<0.01/). 6. Through acute toxicity test, anti-drunk test, anti-inebriation tests on Pinctada martensii glycogen, we study its sober up effect, and then we analyzed its dose-effect relationship. Test results show that when the the oral dose of Pinctada martensii glycogen is 64mg //kg body weight with no side effect. Pinctada martensii glycogen can significantly prolong the tolerance time and short the sorb up time. The best doses of glycogen is 0.80mg//ml. Compared with the saline group, the serum ethanol concentration of Pinctada martensii glycogen group is significantly reduced by 56.63/% /(P <0.01/). 7. By measuring the activity of alcohol dehydrogenase /(ADH/) and aldehyde dehydrogenase /(ALDH/) as an index, we gave the Pinctada martensii glycogen hangover a preliminary study on mechanism of anti-alcoholism. The results show that, compared with the control group, Pinctada martensii glycogen can significantly improve the activity of both ADH and ALDH/(P<0.01/). Compared with the saline group, in those time point we studied, in the glycogen group, the activity of ADH and ALDH rose by an average of 85.46/% and 99.84/%, respectively. It means that, through the promotion of the activity of ADH and ALDH, Pinctada martensii glycogen can rapidly translate the ethanol and acetaldehyde into acetic acid which then passes out of the body, and finally achieve the anti-alcoholism effect.

年份：2010