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摘要：目的：通过测定蚂蟥在冬眠期间体内糖、脂肪、蛋白质代谢相关酶基因的表达，研究蚂蟥冬眠期间物质代谢的机理。方法：选取冬眠0、30、60、90d及正常25℃停食2d蚂蟥的肌肉和消化道，通过实时荧光定量PCR测量相关基因表达。结果表明：消化道中PK、FBP、PEPCK、ACC、HSL、TLSP、AST、GDH基因在冬眠期间的表达呈现先上升后下降的趋势，肌肉中除FBP基因冬眠时期呈先下降、上升、下降的趋势外，其余基因均呈先上升再下降的趋势，且在60d前后达到最高，差异显著（P＜0.05）。结论：蚂蟥在冬眠初期会降低自身代谢，随着冬眠时间的增长，会通过消耗自身物质提供能量。31338
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毕业论文关键词：蚂蟥；冬眠；物质代谢；基因表达
Study on gene expression of the key enzymes in primary metabolism from Whitmania pigra Whitman during hibernation
Abstract： The experimental purpose is to explore the related enzyme gene expression of sugar, fat, protein synthesis to understand the mechanism of W. Pigra metabolism during hibernation.The methods is to select the W. Pigra in different stages of hibernation as material,The gene expression of key enzymes in substance metabolism at different stages was measured by real-time quantitative PCR.The results showed that the expression of PK, FBP, PEPCK, ACC, HSL, TLSP, AST and GDH in the digestive tract increased first and then decreased during hibernation,except for the FBP gene in the muscle during hibernation, it was the first to decrease and increase and decrease, the decrease was the trend of the other genes,It reached the highest level before and after 60 days, and the difference was remarkable（P＜0.05）. The W. Pigra will reduce their metabolism during hibernation early during hibernation with growth time, will provide energy through the consumption of their own material.
Key words:Whitmania pigra Whitman；hibernation；secondary metabolites ；gene expression
目 录
摘要 1
关键词 1
Abstract 1
Key words 1
引言 1
1 材料与方法 2
1．1 材料 2
1．1．1 仪器与试剂 2
1．1．2 实验动物 2
1．2 方法 2
1．2．1 蚂蟥冬眠模型的设计 2
1．2．2 样品的采集与保存 2
1．2．3 RNA提取与cDNA合成 2
1．2．4 引物特异性检测 2
1．2．5 蚂蟥不同部位物质代谢酶基因及抗凝基因表达量的测定 3
1．2．6 数据分析 3
2 结果与分析 3
2．1 RNA提取质量 3
2．2 引物特性检测 3
2．3 冬眠对蚂蟥糖代谢相关酶基因表达量的影响 4
2．4 冬眠对蚂蟥脂肪代谢相关酶基因表达量的影响 5
2．4 冬眠对蚂蟥氨基酸代谢相关酶基因表达量的影响 5
3 讨论 6
3．1 蚂蟥冬眠及冬眠过程中糖的利用 6
3．2 蚂蟥冬眠及冬眠过程中脂肪的利用 6
3．3 蚂蟥冬眠及冬眠过程中蛋白质的利用 7
小结 7
致谢 7
参考文献 8
蚂蟥冬眠过程中糖、脂肪、蛋白质关键酶基因表达的研究
蚂蟥（Whitmania pigra Whitman），水蛭科动物，别名宽体金线蛭，其干燥全体是药材水蛭的主要基原动物，具有破血消症，逐瘀通经的功效[1]，是治疗心脑血管中成药的主要原料。近年来由于野外生存环境污染和市场需求的加大，蚂蟥的供需矛盾日益凸显，人工养殖备受重视。