联合嗅鞘细胞及GDNF的生物可降解膜系统在海绵体神经损伤勃起功能障碍中的作用及机制研究

Erectile dysfunction caused by radical prostatectomy related injury of the cavernous nerve（CN） has a high incidence. This complication can seriously affect the patients' quality of life and cannot be well treated at present. As an specific gliacyte in olfactory system，olfactory ensheathing cells(OECs) play an important role in nerve regeneration. It also be widely used in repair of nerve injury. Glial cell line-derived neurotrophic factor(GDNF) is the most effective neurotrophic factor to promote motor neuron growth in vitro for now. Combining OECs and GDNF show a promising effects in our previous work on repair spinal cord injury and CN ablation. In this research, we attempt to treat rat erectile d ysfunction caused by cavernous nerve injury by Olfactory ensheathing cells and glial cell line-derived neurotropic factor (GDNF) -incorporated PGA/PLA membrane system. And this research will provide some evidences on preventive treatment of this kind of erectile dysfunction. After the animal model used rat is successfully built, a series of experiments will be done. Such as measuring intracavernousal pressure to evaluate the degree of restoring erectile function after cavernous nerve injury. We use electron microscope to observe axon regeneration of injured nerve directly, and to test the function of regenerated nerve fibers, electro-physiological examination and horseradish peroxidase retrograde Tracing will be performed. We use Nissl staining, caspase-3 immunofluorescence staining, terminal deoxynucleotidyl transferase dUTP nick end labeling to observe the survival and apoptosis of neurons in both proximal and distal of injured area. Nitric oxide synthase and neurofilament in cavernosum and dorsal penile nerve, and some specific makers associating to survival or apoptosis of endothelial cells and smooth muscle cells will be showed by immunofluorescence and quantified by west blot. Results from these methods above, may help us discover the mechanisms by which OECs -GDNF- incorporated PGA/PLA membrane system restore erectile function in a rat model of cavernous nerves injury.

前列腺癌根治术中损伤海绵体神经（CN）导致的术后勃起功能障碍（ED）发生率高，严重影响生活质量。现有治疗方法尚未取得满意的效果。嗅鞘细胞(OECs) 是嗅觉系统特有的神经胶质细胞，在神经再生中起重要作用。GDNF是目前发现体外支持运动神经元生长最有效的神经营养因子。在我们前期对脊髓损伤及CN离断修复中，二者联合应用展现出巨大的神经修复前景。本课题拟采用种植有OECs的可降解生物高分子材料-PGA/PLA膜，联合GDNF治疗CN钳夹损伤导致的ED。研究采用大鼠构建动物模型，模拟前列腺癌根治术中CN损伤。采用可降解膜作为联合OECs及GDNF的桥梁。通过评估模型损伤/损伤+修复后的勃起功能、神经元存活、凋亡及轴突再生及功能、海绵体形态学变化等来探索PGA/PLA膜联合OECS及GDNF重建前列腺癌根治术后ED的勃起功能的治疗效果和可能机制并为临床预防性治疗前列腺癌根治术后ED提供新的思路。