Diabetes Erectile dysfunction and apoptosis of cavernous tissues

糖尿病勃起功能障碍与海绵体组织凋亡

• 批准号: 7487368
• 负责人: RAJVIR DAHIYA
• 金额: $ 30.74万
• 依托单位: NORTHERN CALIFORNIA INSTITUTE/RES/EDU
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-15 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7487368
• 关键词: Actins; Affect; Animal Model; Animals; Antibodies; Antioxidants; Apoptosis; Apoptotic; Biochemical; Cell physiology; Condition; DNA Damage; DNA Fragmentation; Daily; Data; Desmin; Development; Diabetes Mellitus; Differentiation Antigens; Dose; Electron Microscopy; Erectile dysfunction; Event; Experimental Diabetes Mellitus; Gene Delivery; Gene Expression; Genes; Goals; Harvest; Hemoglobin; Histology; Hyperglycemia; Hypoglycemia; Immunohistochemistry; In Situ Hybridization; Injection of therapeutic agent; Insulin; Laminin; Lead; Light; Measurement; Mediating; Messenger RNA; Molecular; Myosin ATPase; Nerve; Numbers; Oxidative Stress; Oxidative Stress Pathway; Pathway interactions; Penile Erection; Polymerase Chain Reaction; Principal Investigator; Publications; Rattus; Reverse Transcriptase Polymerase Chain Reaction; Screening procedure; Smooth Muscle; Stream; Streptozocin; Structure of beta Cell of islet; Subcutaneous Injections; Superoxide Dismutase; TdT-Mediated dUTP Nick End Labeling Assay; Techniques; Testing; Time; Tissues; Transfection; United States; Vimentin; Vinculin; Week; adeno-associated viral vector; base; blood glucose regulation; caspase-3; cell type; cytochrome c; day; diabetic; diabetic rat; dosage; gene therapy; glucose monitor; glutathione peroxidase; indexing; intracellular protein transport; men; novel; penis; piperidine; pressure; programs; protein localization location; research study; restoration; treatment duration

DESCRIPTION (provided by applicant): The main goal of this proposal is to investigate whether oxidative stress and apoptosis of cavernous tissues are major pathways in erectile dysfunction. We will also investigate whether inhibition of oxidative stress and apoptosis can restore erectile function. The rationale for this project is that erectile dysfunction affects about 30 million men in the United States and more than 200 million men worldwide. Based on our preliminary data and prior publications, we hypothesize that apoptosis of cavernous tissues are down stream events in erectile dysfunction. We further hypothesize that anti-oxidative agent or Bcl2 gene therapy can restore erectile function in animal model. We will test these hypotheses through the following experiments: Specific Aim# 1. To test the hypothesis that diabetes-induced erectile dysfunction is due to apoptosis of cavernous tissues. Under this specific aim, we will analyze erectile function in diabetic rats and then analyze apoptotic index, anti-apoptotic genes, pro-apoptotic genes, cytochrome c, superoxide dismutase and smooth muscle differentiation markers. We will also investigate whether an anti-oxidative agent (TEMPOL) can restore erectile function in diabetic rats. We will use the following techniques: RT-PCR, real-time RT-PCR, in situ hybridization, immunohistochemistry, electro-stimulation of cavernous nerve for pressure measurements, gene delivery, histology, light and electron microscopy. Specific Aim # 2. To test the hypothesis that insulin treatment can restore erectile function through restoration of anti-apoptotic factors in cavernous tissues of diabetic rats. Under this specific aim, we will treat diabetic rats with insulin and then investigate whether erectile function is restored through restoration of anti-apoptotic pathways described under specific aim # 1. Specific Aim # 3. To test the hypothesis that gene therapy with the Bcl2 anti-apoptotic gene can restore functional, cellular and molecular mechanisms of erectile dysfunction in diabetic rats. Under this specific aim, the Bcl2 gene will be transfected into the cavernous tissue of diabetic rats using an adeno-associated virus vector. We will do the following experiments: (a) Functional assessment of the effects of Bcl2 gene therapy on penile erection in diabetic rats, (b) Analysis of the effects of Bcl2 gene therapy on the oxidative stress pathways and the apoptotic pathways described under specific aim 1. (c) Analysis of the effects of Bcl2 gene therapy on the expression of smooth muscle markers in diabetic rat penis. This proposal is novel because oxidative stress and apoptotic pathways in diabetes-induced erectile dysfunction have never been investigated. Accomplishment of these experiments will provide novel molecular mechanisms of erectile dysfunction and can help guide us towards the development of new therapies for the treatment of erectile dysfunction.

描述（由申请人提供）：该提案的主要目标是研究氧化应激和海绵状组织的凋亡是否是勃起功能障碍的主要途径。我们还将研究抑制氧化应激和凋亡是否可以恢复勃起功能。该项目的理由是，勃起功能障碍影响着美国约3000万人，在全球范围内影响超过2亿人。根据我们的初步数据和先前的出版物，我们假设海绵状组织的凋亡是勃起功能障碍中的溪流事件。我们进一步假设抗氧化剂或BCL2基因治疗可以恢复动物模型中的勃起功能。我们将通过以下实验检验这些假设：特定目的＃1。为了检验糖尿病诱导的勃起功能障碍的假设是由于海绵状组织的凋亡引起的。在这个具体目的下，我们将分析糖尿病大鼠中的勃起功能，然后分析凋亡指数，抗凋亡基因，促凋亡基因，细胞色素C，超氧化物歧化酶和平滑肌分化标记物。我们还将研究抗氧化剂（Tempol）是否可以恢复糖尿病大鼠的勃起功能。我们将使用以下技术：RT-PCR，实时RT-PCR，原位杂交，免疫组织化学，海绵神经的电刺激用于压力测量，基因递送，组织学，光和电子显微镜。具体目的＃2。检验以下假设：胰岛素治疗可以通过恢复糖尿病大鼠海绵状组织中的抗凋亡因子来恢复勃起功能。在这个具体目标下，我们将通过胰岛素治疗糖尿病大鼠，然后研究是否通过恢复特定目的1的抗凋亡途径来恢复勃起功能。特定目的＃3。要测试用Bcl2抗凋亡基因进行基因治疗的假说可以恢复甲基甲基甲基甲基甲基化的功能，细胞和分子机械依次恢复功能性，细胞和分子机制。在这个具体目的下，使用与腺相关的病毒载体将BCL2基因转染到糖尿病大鼠的海绵状组织中。我们将进行以下实验：（a）对Bcl2基因治疗对糖尿病大鼠阴茎勃起的影响的功能评估，（b）分析Bcl2基因治疗对氧化应激途径的影响以及在特定目标下描述的凋亡途径。该建议是新颖的，因为从未研究过糖尿病引起的勃起功能障碍的氧化应激和凋亡途径。这些实验的完成将提供勃起功能障碍的新型分子机制，并可以帮助我们开发新的勃起功能障碍治疗的疗法。