Gene Therapy of Combined Nitric Oxide Synthases and Related Enzymes to Achieve Complete Regression of Advanced Atherosclerosis-Combined Regulation of Substrate of the Enzymes, Co-factors, Transcriptional Factors and Intracellular Enzymes-

背景）我们先前发现，在球囊损伤+高胆固醇饮食诱导的兔动脉粥样硬化中，通过体内基因转移eNOS可以部分消退。(JSPS科学研究补助金（B）项目编号09470166）。目的）探讨eNOS、iNOS及相关酶联合基因转移对动脉粥样硬化（AS）血管充分消退的可能性方法）1）探讨eNOS、iNOS及eNOS+iNOS联合基因转移对AS血管充分消退的可能性。2）成功制备了ERα（雌激素受体α）腺病毒载体。3）制备自发性糖尿病大鼠和老年大鼠的动脉。本实验观察了eNOS、iNOS或eNOS+iNOS基因转染对血管功能的影响。4）阐明了内皮细胞中的酰化和小窝的功能及其与eNOS的关系。 ...更多信息 5）阐明了iNOS在血管功能中的促进或抑制作用。6）探讨eNOS、iNOS或ERα基因联合转染对动脉粥样硬化的治疗作用。结果1）eNOS基因转染可使动脉粥样硬化部分消退，而iNOS或eNOS+iNOS基因转染不能使动脉粥样硬化部分消退。2）制备ERα腺病毒载体，观察其基因转染后NO释放的增加。3）制备自发性糖尿病大鼠动脉和老年大鼠动脉。eNOS基因转染可改善血管功能。4)eNOS的酰化是内皮细胞存活的关键。小窝的形态变化与eNOS活性的变化有关。5)iNOS的siRNA改善血管功能。6）eNOS + ERα联合基因转染可使晚期动脉粥样硬化部分但充分消退，而iNOS不能。结论）eNOS + ERα联合基因转染可使晚期动脉粥样硬化部分但充分消退，而eNOS + iNOS或iNOS + ERα联合基因转染则不能。联合基因转移可能成为实现动脉粥样硬化消退的一种工具。少

Background)We previously found the partial regression by in vivo gene transfer of eNOS in balloon injury + high-cholesterol diet induced atherosclerosis in rabbit. (JSPS Grant-in-Aid for Scientific Research (B) project number 09470166). However, the regression was not sufficient and the mechanism was not well known.Objectives)We investigated the possibility of the sufficient regression by combined gene transfer of eNOS, iNOS and related enzymes in atherosclerotic arteries.Methods)1) We investigated the possibility of the sufficient regression by combined gene transfer of eNOS, iNOS or eNOS+iNOS in advanced atherosclerotic arteries. 2)We produced adenovirus vector of ERα(estrogen receptor α). 3)We prepared the arteries from spontaneously developed diabetic rats and those from senile rats. We investigated the effect of gene transfer of eNOS, iNOS or eNOS+iNOS on their vascular function. 4)We elucidated the function of acylation and caveolae with the relation to eNOS in endothelial cells. … More 5)We elucidated the effect of promotion or inhibition of iNOS in vascular function. 6)We investigated the possibility of the sufficient regression by combined gene transfer of eNOS, iNOS or ERα in advanced atherosclerotic arteries.Results)1)In vivo gene transfer of eNOS, but not iNOS or eNOS+iNOS partially regressed the advanced atherosclerotic arteries. 2)We produced adenovirus vector of ERα and observed the increase of NO release its gene transfer on. 3)We prepared the arteries from spontaneously developed diabetic rats and those from senile rats. We eNOS gene transfer improved their vascular function. 4)Acylation of eNOS is critica for survival of endothelial cell. The morphology of caveolae changes with the relation of eNOS activity. 5)SiRNA of iNOS improved vascular function. 6)We achieved the partial but sufficient regression by combined gene transfer of eNOS plus ERα,but not iNOS in advanced atherosclerotic arteries.Conclusion)Combined gene transfer of eNOS plus ERα,but not eNOS plus iNOS or iNOS plus ERα made possible partial but sufficient regression in advanced atherosclerotic arteries. Combined gene transfer may become one tool to achieve regression of atherosclerosis. Less