金线莲醇提物在斑马鱼中降血糖效果的探究  

Exploration of the hypoglycemic effect of Anoectochilus roxburghii alcohol extracts on diabetic zebrafish models

作　　者：许璟瑾 张文娟[1] 陈志永 李秀敏[1] 潘裕添[1] 王兵丽[1] 欧一新[1,2] 薛钰 XU Jingjin;ZHANG Wenjuan;CHEN Zhiyong;LI Xiumin;PAN Yutian;WANG Bingli;OU Yixin;XUE Yu(The Engineering Technology Center of Mushroom Industry of Minnan Normal University,Zhangzhou 363000,China;School of Life Sciences and Biotechnology,Shanghai Jiao Tong University,Shanghai 200240;State Key Laboratory of Membrane Biology,Beijing 100101)

机构地区：[1]闽南师范大学菌物产业工程技术中心,福建漳州363000 [2]上海交通大学,生命科学技术学院,上海200240 [3]膜生物学国家重点实验室,北京100101

出　　处：《中国比较医学杂志》2018年第6期21-28,共8页Chinese Journal of Comparative Medicine

基　　金：福建省自然科学基金项目资助(2015J05070); 福建省高校杰出青年科研人才培育计划项目; 福建省教育厅科技项目(JK2015028); 福建省中青年教师教育科研项目(JA15318)

摘　　要：目的以斑马鱼为模型筛选金线莲具有降血糖效果的活性组分,并进行初步的机理研究。方法对金线莲进行分离与提取,分成三个组分:金线莲醇提物、大分子多糖(≥5×10~3)和小分子多糖(<5×10~3)。将发育至24 h的斑马鱼胚胎暴露于2%浓度的葡萄糖溶液(2%Glu)中,模拟急性高糖模型,待胚胎发育至48 h时再分别加入三种组分,同时继续用2%葡萄糖溶液处理,72 h时检测不同组分处理后幼鱼的组织液葡萄糖含量,筛选出具有降血糖效果的活性组分。利用半定量PCR和整胚原位杂交技术检测糖代谢相关基因mRNA水平的表达。结果本研究利用高糖胁迫构建斑马鱼糖尿病模型,在该模型下,糖代谢关键因子胰岛素基因(preproinsulin,insulin)、磷酸烯醇式羧基酶1基因(phosphoenolpyruvate carboxykinase 1,pck-1)、胰腺十二指肠同源盒基因1(pancreatic and duodenal homeobox 1,pdx-1)的表达都发生了明显改变,金线莲醇提物能够改善这些基因在高糖胁迫下引起的的异常表达,甚至恢复到正常水平。结论金线莲醇提物对斑马鱼高糖模型具有明显的降血糖功效,该研究为斑马鱼作为糖尿病模型用于探索降血糖药物提供了实验思路和手段。Objective The aim of this study is to explore the hypoglycemic effect of active components ofAnoectochilus roxburghii on zebrafish models. Methods Anoectochilus roxburghii components were extracted and separatedinto three groups： the alcohol extraction group, macromolecular polysaccharide group （≥ 5 × 10^3） and small molecularpolysaccharide group （ 〈 5 × 10^3）. Zebrafish embryos were exposed to 2% glucose solution （2% Glu） at 24 h to imitateacute hyperglycemia phenotype, and then treated with the three Anoectochilus roxburghii components. Based on this high-glucose model, the zebrafish embryos at 72 h were collected to detect the whole tissue glucose value. Furthermore, semi-quantitative PCR and whole mount in situ hybridization were performed to detect the expression of mRNA levels of glycometabolism-related genes. Results An acute diabetic zebrafish model was induced by high glucose stress. In this model,some key factors during glycometabolism such as insulin, pck-1 and pdx-1 were significantly affected, while the alcohol extracts ofAnoectochilus roxburghii obviously reversed these abnormalities induced by high glucose stress, even to normal levels.Conclusions The alcohol extracts of Anoectochilus roxburghii has obvious hypoglycemic effect on diabetic zebrafish model. Ourresult suggest that Anoectochilus roxburghii has a potential application in the hypoglycemic drug screening.

关 键 词：金线莲醇提物 斑马鱼 组织液葡萄糖 高糖模型 糖代谢基因 

分 类 号：R-33[医药卫生]