【作者】 朱威；

【导师】 胡福良；

【作者基本信息】 浙江大学 ， 特种经济动物饲养， 2004， 硕士

【摘要】 工蜂虫蛹自古就是药膳两用食品，我国许多中药药典都有工蜂虫蛹用于疾病治疗以及保健的记录。因其具有较高的开发利用价值，近几年已有一些研究人员对虫蛹的功效重新进行了探讨并取得了一定成果。本实验采用酶联免疫、荧光分光、紫外分光、免疫组化等技术，观察比较工蜂胎的水提物和醚提物对多种炎症动物模型的影响，首次在整体水平、细胞水平、分子水平系统地研究了工蜂胎的抗炎作用及其机理，并对其抗炎免疫成分进行了初步筛选。结果表明： 1．工蜂胎的水提物能够在一定程度上降低炎症区域毛细血管的通透性，能够抑制完全弗氏佐剂引起的炎性肿胀，对角叉菜胶引起的炎性肿胀效果不明显，对油酸加LPS两次打击所致大鼠急性肺损伤有明显的保护作用，能对抗肺水肿的产生；对慢性炎症（棉球肉芽肿模型）具有显著的抑制作用。而醚提物有一定的趋势，但效果并不明显。 2．工蜂胎的水提物能够减少角叉菜胶致大鼠急性胸膜炎炎症组织中PGE₂和总蛋白的含量，降低炎症组织脂质过氧化物代谢产物（MDA）的含量，减轻炎症发生程度，抑制炎症组织中白细胞总数的增加。工蜂胎水提物对大鼠肾上腺中胆固醇的含量和VitC含量影响明显，提示抗炎免疫作用有可能是依赖于垂体-肾上腺皮质系统。但工蜂胎的醚提物对于炎症介质以及胆固醇和VitC含量的影响不大。 3．工蜂胎水提物对急性炎症渗出液中NO及溶菌酶含量的升高表现出抑制作用，能够对抗炎症模型内细胞因子IL-6的异常升高，对IL-2和IFN-γ影响不明显，暗示工蜂胎水提物的抗炎免疫通过作用于B淋巴细胞系统起作用。 4．工蜂胎水提物能够在一定程度上抑制急性肺损伤大鼠支气管上皮细胞中CD54的表达，阻止核转录因子NF-kB p65的活化，抑制大鼠支气管上皮细胞中NF-kB p65 mRNA的表达。提示工蜂胎水提物主要通过抑制炎症细胞中NF-kB p65的活化及其NF-kBp65 mRNA的转录增加控制炎症反应的中心环节，使有害细胞因子的产生减少，保护机体免受有害刺激的进一步损伤，从而达到有效抗炎目的。工蜂胎的醚提物对于CD54的表达、NF-kB p65的活化影响不大。浙江大学硕士学位让走5工蜂胎的功效成分可能主要存在于其水提物中，二蜂胎的水提物呈现出较为明显的抗 炎免疫效果，而醚提物有一定的抗炎趋势，但效果跟工蜂胎的全组分相比较，有一定 的差别。暗示工蜂胎对于免疫方面的作用是一个整体作用，也有可能在分离过程中破 坏了功效成分的原有状态，将其活性成分的药理学效应降低。6.工蜂胎及其水提物和醚提物的急性毒性实验发现，工蜂胎水提物和醚提物的最大耐受 量均在正常剂量的600倍以上，属于无毒物质。用药期间对于胸腺等免疫器官影响不 明显，并且能够对抗炎症造模期间的体重减轻。 同时，对工蜂胎的开发利用的研究方向进行了讨论，认为工蜂胎今后的研究方向是 加工工艺、保藏方式以及药物作用机理方面的研究为主。本实验结果为工蜂胎的开发利 用提供了理论依据。更多还原

【Abstract】 Worker honeybee larvae and pupae (WHLP) are used for meals and medicines for its well-known biological activities since the ancient time. Compendium of Materia Medica and other Chinese traditional pharmacopoeia recorded the application of WHLP for treatment and health care. Recently some researchrers made a new sight into WHLP for its high economic value. This experiment determined the anti-inflammatory effect of water extraction (WE) and ether extraction (EE) of WHLP through enzyme-linked immunosorbent assay, fluoro-spectrophotometry, ultraviolet-spectrophototometry, immunohistochemistry and in situ hybridization at the level of the cell and molecules. The results are as follow:1. WE could debase the permeability of capillary vessel of inflammatory tissue in some degree, and show a complete inhibitory effect on the edemas induced by Freund’s adjuvant complete and there is no obvious inhibitory effect found on edemas induced by carrageenan; WE indicates an obvious protective effect on the "twice-hit" animal model of acute lung injury in rats induced, by oleic acid and lipopolysaccharide (LPS) from lung edema. It also shows an excellent inhibition function on the chronic granuloma as well. EE shews indistinctive effect.2. WE can decrease the content of inflammatory mediators (PGEi and total protein), total protein and lipid oxide products (MDA) in inflamed tissue, and alleviate the inflammatory response and reduce the amounts of leukocytes. The obvious effect of WE and indistinctive effect of EE on the contents of cholesterol and VitC in adrenal glands suggestes that the inhibition effects on inflammation tissue of WE may depend on the pituitary-adrenal system.3. WE can inhibit the abnormal increase of NO, lysozyme and cytokines IL-6 in acute inflammatory tissue fluid. It shows no significant effect on the content of cytokines IL-2 and IFN- Y . This suggests that WE act through B lymphocyte system.4. WE can inhibt the expression of CD54 in the bronchial epithelium tissue and prevent the activation of NF-kB p65 and transcriptiion of mRNA of NF-kB p65 in the bronchialepithelium tissue of acute lung injured rats. It protects the tissue from farther injury by controlling the key step to inflammatory response. EE doesn’t show a similar result.5. The main acting components of WHLP could be possibly extracted by water. WE shows an obvious anti-inflammation effect while EE is not conspicuous for its inhibitory effect, and both of them isn’t as effective as the whole WHLP. It suggests that it may be the resultant effect of whole WHLP for anti-inflammation, and the process of extraction destroys some active ingredients of WHLP.6. Acute toxicity testing of WE and EE demonstrate that the maximal tolerance dose exceeds the normal dose more than 600 times, which can be considered as nontoxicity. There are no significant effects on immune organ in the course of giving drugs, and they can antagonize body weight loss when creating the model.At the same time, the research direction of exploitation and development are discussed. Future aim of WHLP is to study the process technique and its pharmacology mechanism. The research results provide scientific basis for the application of WHLP as health food and medicine use.更多还原