运用家蚕糖尿病模型和切除HPLC法确认壮药酸荔枝中的降糖成分

Silkworm as drug screening in animal models has many advantages, Sekimizu Kazuhisa, a professor at University of Tokyo in Japan, reported that the total sugar in the silkworm was the index, successfully established for the establishment of type II diabetic animal model. The establishment of type II diabetic animal model by using the silkworm blood trehalose as index is feasible, the Shandong University scholar Professor Cui Weizheng reported. But no reports at home and abroad, which study Chinese medicine research reported in the silkworm as a valid model of diabetes and by knock out HPLC. Our group was found in the silkworm blood glucose as an indicator of Type II diabetes model can be established, to be drug pharmacokinetics and immunoblot analysis of its mechanism to make a detailed study, the main test silkworm phosphorylated antibodies in the blood Akt, phospho-JNK, phosphorylation of AMPK and β-actin protein expression. And on this basis, we intend to preclude the use of different extraction purification and other separation methods to identify the best wild lychee high medicine hypoglycemic activity of acid. Then take the current knock out HPLC method to find lead compounds. Initially establish a hypoglycemic drug screening of low cost animal model for further study on the hypoglycemic herbs, to provide a new way.

以家蚕作为药物筛选模式动物具有诸多优势, 如日本东京大学教授Kazuhisa Sekimizu 报道以家蚕血液总糖为指标成功的建立了二型糖尿病动物模型。山东大学学者崔为正教授报道以家蚕血液海藻糖为指标建立二型糖尿病动物模型是可行的。但是国内外未有以家蚕血液游离葡萄糖为指标作建立糖尿病模型和切除HPLC法来研究中药有效成分研究的报道。本课题组预实验发现以家蚕血液游离葡萄糖为指标的二型糖尿病模型是可以建立的，拟用药代动力学和免疫印迹分析法对其机理做一个详细的研究，主要检测家蚕血液中抗体磷酸化Akt、磷酸化JNK，磷酸化AMPK和β肌动蛋白的表达。并在这个基础上拟釆用不同分离萃取提纯等方法, 找出野生壮药酸荔枝最佳的降糖活性混合物。 进而采取切除HPLC法来找出有前景的化合物，初步建立一种低成本的降糖药物筛选动物模型, 为今后研究降糖中草药提供一种新的思路。

采用广西本地蚕种, 以本地桑叶为饲料建立成功稳定的家蚕糖尿病模型，建模成功率在99%以上。采用以上模型对广西酸荔枝提取物进行了降糖效果评价，发现了两个单体化合物降血糖效果良好，分别是Quercetin hydrate和Qurecetin-3-beta-D-Glucoside。这确实证明了此模型具有应用价值，将来可以用来对中草药降血糖活性成分进行初筛。. 出现的意外情况：采用切除HPLC法，对酸荔枝提取物的降糖活性部位进行筛选，发现由于蚕的个体差异有些偏大，很难确定。目前我们采用的方法尽量是选择同年龄同大小进一步打算想选同性别的蚕进行精准建模，将来可能解决这个问题。. 同时在本课题的研究中还意外的发现.1利用糖尿病家蚕可以建立糖尿病足模型，并且此模型的成功率目前为65%。2 铬的有机配合物在降血糖方面有良好的效果，并在蚕糖尿病模型上进行了验证。3发现苹果酸铬可以诱导家蚕吐丝直接成蚕丝被。

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