经皮电刺激对大鼠海绵体神经损伤后nNOS表达的研究

发布时间：2018-09-05 16:14

【摘要】： 【研究背景】阴茎勃起功能障碍(erectile dysfunction,ED)是最常见的男性性功能障碍,法医学领域在人身伤害、交通事故、离婚、强奸、医疗纠纷等方面都会涉及阴茎勃起功能的鉴定。虽然通过夜间勃起功能监测(NPT)、彩色双功能多普勒(CDDU)、阴部诱发电位等方法可以对阴茎勃起功能进行鉴定,但为了深入研究ED的发生机制,并探讨ED分子水平可能的诊断标准,必须建立动物模型。作为支配阴茎勃起的最重要神经,海绵体神经是建立神经性ED模型的首选神经,而大鼠在盆腔、阴茎的血管及神经解剖等方面都与人有较强的相似性,而且其价格也是ED研究的动物中最适宜的,因此大鼠海绵体神经损伤是ED研究的理想实验模型。神经性ED的诊断技术虽然取得了很大进步,但神经性ED的治疗仍然是十分困难的,目前尚无有效的治疗方法。电刺激是一种传统的神经康复疗法,可能通过促进生长因子的趋化作用、增加雪旺细胞的活性等促进神经再生。一氧化氮(NO)是阴茎勃起的关键性神经递质,通过舒张阴茎海绵体平滑肌细胞而引起阴茎勃起,一氧化氮合酶(NOS)是合成NO的关键酶。通过神经性一氧化氮合酶(nNOS)检测可以反映电刺激后海绵体神经功能恢复的情况,间接反映阴茎的勃起功能。【目的】本实验拟建立SD大鼠双侧海绵体神经钳夹伤模型,观察伤后勃起功能及nNOS的改变,并定时定量给予经皮电刺激后观测nNOS恢复的情况。为海绵体神经损伤所致ED的法医学鉴定提供分子水平依据,也为临床上神经性ED的治疗提供一种可能的方法。【材料与方法】将30只SD成年雄性大鼠分成实验组和对照组,建立海绵体神经损伤致神经性ED模型。然后另取30只大鼠分为实验组和对照组,按海绵体神经损伤后是否给予经皮电刺激,将实验组分为2组(损伤后定时、定量给予经皮电刺激组10只;损伤后未给予任何干预措施组10只);对照组为正常SD大鼠,10只,未作任何处理。通过电刺激海绵体神经观察其勃起现象,一个月后取阴茎中段组织,切片,光镜观察大鼠阴茎海绵体的解剖结构,免疫组化方法检测nNOS表达的情况。分析海绵体神经损伤后nNOS表达变化及经皮电刺激后nNOS恢复情况。【结果】通过电刺激反应证实造模成功,损伤组与对照组有显著性差异。在切片上观察任意4个高倍视野(400×),计数视野中nNOS阳性神经纤维灰度和吸光度,损伤后未给经皮电刺激组198.12±3.27和6.12±1.17,损伤后给予经皮电刺激组191.27±4.20和7.68±1.72,正常对照组分别为180.39±3.77和8.25±1.03。损伤后未给经皮电刺激组与对照组nNOS阳性神经纤维灰度和吸光度存在显著性差异(P0.05),损伤后未给经皮电刺激组与经皮电刺激组间nNOS阳性神经纤维灰度和吸光度存在显著性差异(P0.05)。【结论】海绵体神经位于前列腺的后外侧,通过双侧钳夹海绵体神经,可以导致勃起功能的丧失,即神经性ED。海绵体神经损伤是建立神经性ED模型的良好方法,通过对盆神经的电刺激实验可以证实模型是否成功。损伤后定期给予定时定量的经皮电刺激可以促进海绵体神经功能的恢复,这可以通过电刺激盆神经观察勃起情况证实,也可以通过nNOS定量变化来反映。该研究对于实际检案中合并海绵体神经损伤所致的ED有一定指导性,也为临床研究海绵体神经再生提供了依据。鉴于国内外有大量海绵体神经损伤所致的神经性ED的现状,为进一步研究其病理生理机制、寻找切实有效的治疗方法提供了重要的依据。
[Abstract]:[BACKGROUND] Erectile dysfunction (ED) is the most common male sexual dysfunction. Forensic medicine involves the identification of erectile dysfunction in the fields of personal injury, traffic accidents, divorce, rape, medical disputes and so on. Partial evoked potentials and other methods can be used to identify the erectile function of the penis, but in order to further study the mechanism of ED and to explore the possible diagnostic criteria of ED molecular level, animal models must be established. The blood vessel and nerve anatomy are similar to human beings, and the price is the most suitable animal for ED study. Therefore, the rat cavernous nerve injury is an ideal experimental model for ED study. Electric stimulation is a traditional neurorehabilitative therapy that promotes nerve regeneration by promoting the chemotaxis of growth factors and increasing the activity of Schwann cells. Nitric oxide (NO) is the key neurotransmitter in penile erection, which causes erection and nitric oxide synthase by relaxing the smooth muscle cells of the cavernous body. Nitric oxide synthase (NOS) is the key enzyme for NO synthesis. The detection of neuronal nitric oxide synthase (nNOS) can reflect the recovery of cavernous nerve function after electrical stimulation, and indirectly reflect the erectile function of penis.
[Objective] To establish a model of bilateral cavernous nerve clamp injury in SD rats, observe the changes of erectile function and nNOS after injury, and observe the recovery of nNOS after percutaneous electrical stimulation regularly and quantitatively. A possible method.
[Materials and Methods] Thirty adult male SD rats were divided into experimental group and control group to establish the neurogenic ED model induced by cavernous nerve injury. 10 rats in group A, 10 rats in group B, 10 rats in group B and 10 normal SD rats in group B were treated without any intervention. Changes of nNOS expression after cavernous nerve injury and nNOS recovery after transcutaneous electrical stimulation.
[Results] The model was successfully established by electrical stimulation, and there was a significant difference between the injured group and the control group. 0 and 7.68 + 1.72, respectively. The normal control group was 180.39 + 3.77 and 8.25 + 1.03. respectively.
There were significant differences in gray scale and absorbance of nNOS-positive nerve fibers between the non-percutaneous electrical stimulation group and the control group (P 0.05). There were significant differences in gray scale and absorbance of nNOS-positive nerve fibers between the non-percutaneous electrical stimulation group and the percutaneous electrical stimulation group (P 0.05).
[Conclusion] The cavernous nerve is located in the posterolateral part of the prostate. The loss of erectile function can be caused by bilateral clamping of the cavernous nerve. Quantity of transcutaneous electrical stimulation can promote the recovery of cavernous nerve function, which can be confirmed by the observation of pelvic nerve erection and can also be reflected by the quantitative changes of nNOS. Basis. In view of the present situation of neurogenic ED caused by a large number of cavernous nerve injuries at home and abroad, it provides an important basis for further study of its pathophysiological mechanism and for finding effective treatment methods.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：D919

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