INTESTINAL MUCOSAL FUNCTION IN DIABETES

大鼠未加控制的糖尿病导致多发性 毛刷边缘膜功能的改变 肠细胞包括主动转运的刺激 膜结合力的各种机理及增加 水解酶。对涉及的机制的阐明可能会提供 对人类疾病病理生理学的重要洞察 以及对刷子边框性质的规定 蛋白质。我们的长期目标是澄清这些 机械装置。我们的具体目标是回答以下问题 问题：(1)蔗糖酶的生物合成速度是否增加- 糖尿病动物体内异麦芽糖酶的转录， 翻译，或翻译后的变化；(2)是变化 是否特定或更改了所有画笔边框的规则 水解酶；(3)调节变化的介体是什么；(4) 在其他情况下水解酶的变化是否与 糖尿病的变化。为了回答这些问题，我们将研究 糖尿病和其他情况下水解酶活性的影响 在前体的合成和细胞内处理上发生改变 选定的笔刷边缘水解酶。这将通过以下方式完成 脉冲/追逐实验之后是免疫沉淀和 十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法和荧光法 免疫电子显微镜。对信使核糖核酸水平的影响 在无细胞翻译实验中寻求的，以及 器官中生物合成过程的几种可能的介体- 肠道培养结果确定。最后，由于细胞内 对这些蛋白质的加工还不完全了解，我们会的， 在这些实验的过程中，更清楚地定义 在正常大鼠肠道中的加工，在几种情况下 水解酶，人体肠道。

Uncontrolled diabetes mellitus in the rat results in multiple alterations in the function of the brush border membrane of the enterocyte including the stimulation of active transport mechanisms and increases in a variety of membrane-bound hydrolases. Elucidation of the mechanisms involved may provide important insight into the patho-physiology of the human disease as well as into the nature of the regulation of brush border proteins. Our long term objective is to elucidate these mechanisms. Our specific aims are to answer the following questions: (1) is the increased rate of biosynthesis of sucrase- isomaltase in diabetic animals the result of transcriptional, translational, or posttranslational changes; (2) are the changes specific or is there altered regulation of all brush border hydrolases; (3) what are the mediators of altered regulation; (4) are alterations in hydrolases in other situations related to the diabetic changes. To answer these questions we will study the effects of diabetes and other conditions where hydrolase activity is altered on synthesis and intracellular processing of precursors of selected brush border hydrolases. This will be done by pulse/chase experiments followed by immune precipitation and SDS-polyacrylamide gel electrophoresis and fluorography and by immunoelectron microscopy. Effects on mRNA levels will be sought in cell free translational experiments, and the effects of several possible mediators on biosynthetic processes in organ- cultured intestine determined. Finally, since intracellular processing of these proteins is incompletely understood, we will, in the course of these experiments, define more clearly this processing in normal rat intestine and, in the case of several hydrolases, human intestine.