【作者】 常帅；

【导师】 郭亚平；

【作者基本信息】 山西大学 ， 动物学， 2008， 硕士

【摘要】 本文用来自蜜蜂分泌的蜂王浆喂养果蝇,研究和检测蜂王浆对果蝇生长发育、寿命和老化的的影响,并且探讨了造成影响的原因。天然来源的许多成分复杂的物质,经常被用来预防和治疗疾病。这些物质多数来自植物,少数来自动物和真菌。正因为成分是混合的,给分析其机理带来不少困难。这可以从两个方面给以解决,一方面用这种混合物在动物上进行试验,看是否有某种功效,就会为人类使用这些物质作一参考。另一方面,将混合物所含成分进行解析,可以是分离检测或根据已有数据的分析,来确定主要成分的多少,将各主要组分的确定功能叠加起来,就能大体上看出这一混合物的功能如何。我们研究蜂王浆就是沿着这个思路进行的。首先做的是果蝇培养和生活史观察,熟悉其生活习性,为以后的实验打下基础。我们在参阅已有文献的基础上,作了较为详细的观察。先前设想蜂王浆既然可以改变蜜蜂的生长、发育和寿命,也将在果蝇中得以体现。随后用培养瓶进行了羽化以后的寿命实验,将果蝇的雌蝇和雄蝇在含蜂王浆与不含蜂王浆的培养基分别培养,观察和记录果蝇的死亡情况,以存活率对时间作图,得出果蝇生存的趋势线,比较出蜂王浆有无延长果蝇寿命的效果。结果发现,蜂王浆延长了雌蝇的平均寿命,没有延长雄蝇的平均寿命,并且曲线的形状在用蜂王浆喂养的实验组中有所变化。可知蜂王浆的确对果蝇寿命有所影响。接下来,用培养管做了果蝇从产卵到羽化的实验,测试蜂王浆对果蝇幼虫的生长影响。产卵后即在实验组加入蜂王浆,对照组未加蜂王浆,然后观察和记录果蝇羽化时间,在加入与不加蜂王浆实验的基础上,并且改变了酵母的含量,在蜂王浆和酵母间也作一比较。从开始羽化后的几天时间里,比较了开始羽化的时间和羽化量最多的一天。发现蜂王浆的确使羽化时间和羽化量最多时间提前,同时酵母也有一定作用。可知蜂王浆对果蝇生长有所影响。后来的实验研究了果蝇体内酶活性和丙二醛含量的高低。设想蜂王浆既然能表现为改变生长与寿命的作用,也必然可以改变果蝇体内的生化指标,即酶活性及其它小分子的量的高低。用培养瓶培养羽化后的果蝇,每隔一段时间每瓶取出存活的果蝇25只,余者继续培养。测定不同时间收集的果蝇体内SOD、CAT比活性和MDA含量,在添加蜂王浆培养的果蝇与未添加的培养果蝇间作比较。用的是果蝇的匀浆后的上清液。结果确实发现了一定的差异。与寿命的变化基本相符合,从而进一步证实了我们的预想。可能的机制是,蜂王浆的加入改变了抗氧化酶活性,然后由SOD和CAT去减少MDA的含量。在解释结果时使用已有资料进行蜂王浆成分剖析,把组分与生理意义对应起来,找出产生结果的原因是蜂王浆有某些成分,而某些成分又恰好具有这些作用,使果蝇的生长、发育和老化状况得到改变,可能也改变了生长、发育和老化过程的激素信号。蜂王浆含有的王浆蛋白质、肽类、激素、酶、脂肪酸和多量维生素等可能起到重要作用。这可为蜂王浆的生产和利用提供一定的参考依据。在前言中介绍前人已取得的研究成果,介绍蜂王浆的部分研究结果,介绍生长发育尤其是老化的有关内容,并对模式生物果蝇做了基础介绍,包括果蝇的遗传操作。为了能在分子水平上开展工作,我们对于果蝇基因的染色体定位克隆方法,以及用到的一些遗传操作方法作了认真的介绍,给出典型的实例,包括抗氧化酶过氧化氢酶的例子,以利于今后同行开展类似工作。更多还原

【Abstract】 Royal jelly secreted from the honey bee is used to feed on fruit flies and test the impact of Royal jelly on growth and development, lifespan and aging of fly, and to explore the reasons which cause these impacts. Many natural sources of material with complex components are often used to prevent and treat disease. Most of these substances come from plants, minorities of these come from animals and fungi. It is precisely because the ingredients are mixed, so bring a lot of difficulties in analysising its mechanism. This can be given in two ways to solve. On the one hand, we use this mixture in the test conducted on animals to see if it have a certain effect, Once it have, result will be as reference for human use of these substances. On the other hand, we do analysis to composition contained in the mixture, it may be separated and examinated or tested according to the analysis of available data, to determine how much of major components, if we stack up the major components of certain functions, we can see what about the functions of the mixture generally. We have studied royal jelly and are conducted along this thinking.First of all we did was to culture fruit flies and observe theirs life history, and get familiar with their living habits, to lay the foundation for future experiments. We have done a more detailed observation based on some references.As previously, since Royal jelly can change the honey bee’s growth, development and life, will also be reflected in the fruit fly. Followed by lifespan experiment used cultivating bottle after the eclosion of fly, the female flies and male flies of fruit flies were cultured respectively in the culture medium that without royal jelly or with royal jelly and the death situation of the fly are observed and recorded. We then mapped with survival rate to time and get the survival trend line of the fruit fly, compared whether royal jelly had result that can extend the life of Drosophila. The results showed that royal jelly extend the average life expectancy of the female flies, but not extend the average life expectancy of male flies, and the shape of the curve changes in the experimental group feeding with royal jelly. It can be known that royal Jelly really affect the lifespan of the fruit fly.Next, using the cultivating tube, we have done the experiment of the flies emerge from the eggs to eclosion, testing of royal jelly on the larvae growth of the fruit fly. After spawning the royal jelly was added to the experimental group, but royal jelly was not added to the control group. Then the emergence times of flies were observed and recorded, On the basis of experiment royal jelly was added to or was not added to culture medium, and we change the content of the yeast, there also was a comparison between royal jelly and yeast. From the beginning after the emergence, until a few days later, we compared the initiatory emergence time and the day that emergence number is most. We find that royal jelly indeed advance the emergence time and the time emergence number is most, and yeast also have a role. It is known that royal Jelly have an impact on the growth of the fruit fly.Later experiments were turned to research degree of activities of the enzyme and the MDA content in fruit flies. Since the royal jelly can performance effect of changing the growth and lifespan, it also can inevitably change the biological chemistry indicators in the body of fruit flies that is the degree of activity of enzymes and content of other small molecules. We culture flies with cultivating bottle after their eclosion, taking out twenty five fruit flies within survival whenever after a period of time from each bottle, the remaining flies are continued to culture. Determining SOD specific activity and the content of MDA of collections of fruit flies on different time, we compared cultured fruit flies between which royal jelly was added in the culture medium and which royal jelly was not added in the culture medium, using the fruit fly’s supernatant after homogenate. The results did find that a certain difference, and have fundamental conformity with lifespan changes, which so further proved what we have expected. Possible mechanism is that the addition of royal jelly has changed antioxidant activity, and then by the SOD and CAT to reduce the content of MDA. For explaining the results, information has been used to analysis components of royal jelly, we set out to find corresponding relationships between the components and physiological significances of royal jelly, and to find out the outcome of the reasons is that there are some components in royal jelly, and because just some components have these roles, so that the growth, development and aging state of fruit fly are changed, may it have changed hormone signal of the growth, development and aging processes. Royal jelly containing the Royal jelly protein, peptides, hormones, enzymes, fatty acids, some kinds of vitamins with high levels, and so on may play an important role. These results can provide a frame of reference for the production and use of royal jelly.In the foreword, we introduce predecessors’ research results that have been made on the part of the research findings about Royal jelly as well as on growth and development especially on the ageing-related contents, and also have introduced basic information on model organism fruit fly, including generic operation of fruit fly. In order to carry out work on the molecular level, we have serious introduction for chromosome location cloning method of the genes in fruit fly and genetic manipulation that could be used. Typical practical examples are given of this method, including example of the antioxidant enzyme catalase, to facilitate the similar work in future.更多还原