C3G－SOD双靶点阻遏ROS多环节改善PINF的实验研究

项目名称： C3G－SOD双靶点阻遏ROS多环节改善PINF的实验研究

项目编号： No.30872531

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 吴育连

作者单位： 浙江大学

项目金额： 30万元

中文摘要： 亲体器官移植可缓解供体紧缺、减少排斥。亲体胰岛移植发展限制的主要原因是胰岛获取及移植后原发性移植物无功能－PINF。我们新近发现C3G 可通过抑制ROS 氧化损伤而明显改善供体鼠胰岛功能；但通过单一C3G 竞争结合自由基可能还不足有效清除ROS 对供体胰岛的损伤，同时结合ROS 酶解激活可能对C3G 有互补协同作用。本项目首次设想：在对杨梅来源的C3G 进行定量分析纯化基础上，利用C3G 在体内外预处理供体、移植物，研究其对胰岛功能及分离纯化产量的影响；同时体外构建稳定表达SOD 基因的供体胰岛，应用C3G 竞争结合自由基及SOD 对内源性ROS 的酶解清除作用，在已建立的临界剂量胰岛移植模型上，通过研究C3G 和SOD 基因转染技术阻遏ROS 不同靶点和PINF 发生的不同环节的作用及其协同效应，探讨其提高胰岛产量和活力及预防PINF 的效率，以达到减少移植供受体比例和移植成本的目标，为推动亲体胰岛移植奠定实验基础。

中文关键词： 胰岛移植物功能；ROS；C3G；SOD

英文摘要： Living islet transplantation could palliate the stress of donor shortage and reduce the risk of graft rejection. However, it is circumscribed due to the limited islet harvest and primary islet nonfunction (PINF). We recently found that cyanidin-3-O-glucoside(C3G) could improve the islet function both in vitro and in vivo studies, which was mainly attributed to the alleviation of ROS-mediated oxidative injury. While the protective effect with single agent is limited, and we presume that activation of intracellular ROS enzymolysis may produce synergistic effect. Thus, the present project firstly hypothesis and focus on the following themes: 1) the effect on the function and quantity of graft islets after treated with purified C3G extracted from Chinese bayberry both in vivo (to donar before islets isolation) and in vitro (to graft islets during culture); 2) we transfect primary islets with SOD gene to enhance the ability of intracellular ROS enzymolysis and to observe whether it could greatly improve the viability and function of graft islets and could effeciently prevent the occurrence of PINF when in combination with C3G treatment for scavenging ROS extracellularly, since these two interventions are targeting different pathways. If so, the present experiment could obviously lower the ratio of donar to recipient as well as the transplantation cost, and may play a role in promoting clinical islet transplantation.

英文关键词： islet graft function;ROS；C3G；SOD