【作者】 李娜；

【导师】 金丽华；

【作者基本信息】 东北林业大学 ， 遗传学， 2019， 硕士

【摘要】 胰岛素抵抗是由于胰岛β细胞所分泌的胰岛素不能发挥正常作用而引起的代谢紊乱。胰岛素信号通路是调节胰岛素代谢的直接途径。研究表明,通过自噬清除受损的细胞器以及错误折叠的蛋白质等,对调节β细胞的数量及结构起到关键性作用。因此,由于饥饿应激条件下造成的代谢紊乱以及细胞自噬之间的关系成为研究热点。果蝇的胰岛素信号通路通过营养条件调控细胞的增殖与生长,并最终实现细胞代谢和细胞大小的相互统一,营养均衡吸收会促进合成代谢、细胞生长、细胞循环;反之,饥饿则会引起相反的作用。中药具有来源广泛、历史悠久、作用范围较大、价格较低廉等优点。为了探究中药对饥饿诱导的果蝇胰岛素信号的影响,本实验通过对果蝇的爬行能力、生长速度以及饥饿生存率分析,从150种中药中筛选了五味子、元胡、茵陈、竹茹、降香、菖蒲、杜仲、山慈菇、女贞子、白鲜皮10种效果明显的中药。为了进一步探究这10种中药对胰岛素信号通路以及细胞自噬的影响,分析了喂食中药后饥饿条件下的脂滴大小、细胞核面积、p62的积累水平、mCherry-Atg8a的表达量、PH3+细胞数量以及Chico¹/CyO、Akt1/Q/TM3.Sb两种胰岛素通路突变体的生存率。实验结果表明在饥饿诱导的条件下,五味子、元胡、茵陈、降香、竹茹、菖蒲能够显著增加脂肪油滴面积;五味子、元胡、茵陈、竹茹、降香、菖蒲、山慈菇、白鲜皮增加p62的积累并降低mCherry-Atg8a表达量;五味子、元胡、茵陈、竹茹、杜仲增加PH3阳性细胞的数量。另外,五味子、元胡、茵陈、降香、竹茹、菖蒲、杜仲、山慈菇、女贞子、白鲜皮能够显著提高Chico¹/CyO和Akt₁Q/TM3.Sb胰岛素通路突变体果蝇饥饿后的生存率。因此,我们筛选的10种中药在一定程度上能够缓解饥饿诱导果蝇的胰岛素代谢紊乱和细胞自噬的影响,此结果能够为中药的综合开发与利用提供理论基础。更多还原

【Abstract】 Insulin resistance is a metabolic disorder caused by the dysfunction of insulin,which is secreted by islet β cells.The insulin signaling pathway is a direct pathway for the regulation of insulin.It has been reported that the clearance of damaged organelles and misfolded proteins by autophagy plays a key role in regulating the number and structure of β-cells.Therefore,increasing attention has been paid to the study of the relationship between metabolic disorders and autophagy caused by starvation stress.The insulin signaling pathway in Drosophila regulates organ metabolism and cell size through nutrient conditions;this regulation is mainly achieved by consistent regulation of cell proliferation,growth and metabolism.Balanced absorption of nutrients promotes anabolism,cell growth,and cell cycle;on the contrary,deprivation of food can cause the opposite effect.Traditional Chinese herbal medicine has the advantages of wide source,long history and low price.To explore the effects of traditional Chinese medicine on starvation-induced insulin signaling pathway in Drosophila,we analyzed the status of growth and development of Drosophila reared in 150 traditional Chinese medicines by examining crawling ability,growth rate and starvation survival rate.Among these,10 kinds of traditional Chinese medicines:Schisandra chinensis（Turcz.）Baill,Corydalis yanhusuo W.T.Wang,Artemisia capillaris Thunb，Bambusa tuldoides Munro,Dalbergia odorifera T.chen,Acorus calamus L,Eucommia ulmoides Oliv,Cremastra appendiculata（D.Don）Makino,Ligustrum lucidum Ait,Dictamnus dasycarpus Turcz were selected for further analysis.To explore the effects of these 10 Chinese medicines on insulin signaling pathway and autophagy,we analyzed the lipid droplet size,nuclear area,p62 accumulation level,mCherry-Atg8a expression,PH3+ cell count and the survival rate of Chico¹/CyO,Akt₁Q/TM3.Sb mutants under starvation conditions.The results showed that:Schisandra chinensis（Turcz.）Baill,Corydalis yanhusuo W.T.Wang,Artemisia capillaris Thunb,Bambusa tuldoides Munro，Dalbergia odorifera.chen,Acorus calamus L and Eucommia ulmoides Oliv can significantly increase the area of fat droplets under the starvation;Schisandra chinensis（Turcz.）Baill,Corydalis yanhusuo.W.T.Wang,Artemisia capillaris Thunb,Bambusa tuldoides Munro,Dalbergia odorifera T.chen,Acorus calamus L,Cremastra appendiculata（D.Don）Makino,Dictamnus dasycarpus Turcz increased the accumulation of p62 and decreased the expression of mCherry-Atg8a.Schisandra chinensis（Turcz.）Baill,Corydalis yanhusuo W.T.Wang,Artemisia capillaris Thunb,Bambusa tuldoides Munro,Eucommia ulmoides Oliv increased the number of PH3 positive cells.In addition,Schisandra chinensis（Turcz.）Baill,Corydalis yanhusuo W.T.Wang,Artemisia capillaris Thunb,Bambusa tuldoides Munro,Dalbergia odorifera T.chen,Acorus calamus L,Eucommia ulmoides Oliv,Cremastra appendiculata（D.Don）Makino,Ligustrum lucidum Ait,Dictamnus dasycarpus Turcz can significantly improve the survival rate of Chico¹/CyO and Akt₁Q/TM3.Sb insulin pathway mutants of Drosophila.The 10 traditional Chinese herbal medicines can alleviate starvation-induced insulin metabolism and autophagy in Drosophila to some extent.These data can provide a theoretical basis for the comprehensive development and utilization of traditional Chinese herbal medicine.更多还原