生殖股神经生殖支移位海绵体神经修复大鼠神经源性勃起功能障碍的研究

发布时间：2018-06-05 06:41

本文选题：神经移位 + 生殖股神经生殖支　；参考：《华中科技大学》2014年博士论文

【摘要】：目的：探讨生殖股神经生殖支移位海绵体神经后,再生神经形态学特点及新建立的勃起反射传出通路的构成。方法：将30只成年雄性Sprague-Dawley大鼠(约250-300g)随机分为三组,每组10只,假手术组：仅显露双侧生殖股神经及海绵体神经,不做离断。损伤组：离断双侧生殖股神经生殖支及海绵体神经,断端以丝线结扎防止再生。神经移位组：离断双侧生殖股神经生殖支及海绵体神经后,双侧生殖股神经生殖支近端与海绵体神经远端行端端吻合。术后12周,将荧光金溶液(Fluorogold,FG)分别注入各组大鼠右侧阴茎脚内行神经逆行示踪研究；再通过各组大鼠远端海绵体神经半薄切片有髓神经纤维计数及电镜观察来评估神经再生情况,其中神经移位组大鼠吻合口段神经进一步行纵切面HE染色,以了解吻合口段神经的连续性。结果：逆行神经示踪发现：损伤组和神经移位组大鼠盆神经节内仅见极少量的FG阳性神经元,其平均数量分别为6.3±3.4和5.4±2.9,远少于假手术大鼠(106.7±15.9)；在各节段的脊髓切片中,假手术组和损伤组大鼠均未发现有FG阳性神经元存在,而神经移位组大鼠脊髓L1,L2前角内可见有FG阳性神经元,其平均数量分别为2.7±1.0,5.3±1.2。神经横断面半薄切片甲苯胺蓝染色显示：假手术组的海绵体神经和神经移位组的再生神经内均可见丰富蓝染的有髓神经纤维,其数量分别为346.8±17.66和621.87±81.25,明显高于损伤组远端海绵体神经内有髓神经纤维的数量(16.68±6.08)(P0.05)；超微电镜进一步观察到假手术组和神经移位组有大量有髓和无髓神经纤维存在,几乎未见变性坏死的髓鞘；而损伤组可见髓鞘广泛变性,胶原纤维大量增生。神经移位组大鼠吻合口段神经HE染色见生殖股神经生殖支可以通过吻合口长入海绵体神经。结论：生殖股神经生殖支移位海绵体神经后,生殖股神经生殖支可以再生长入阴茎海绵体,并建立新的勃起反射传出神经通路(脊髓L1,L2前角运动神经元—生殖股神经生殖支—再生神经—阴茎海绵体)。目的：运用神经电生理学和行为学方法,评估大鼠生殖股神经生殖支移位海绵体神经后阴茎勃起功能的恢复情况。方法：将30只成年雄性Sprague-Dawley大鼠(约250-300g)随机分为三组,每组10只：假手术组,仅显露双侧生殖股神经及海绵体神经,不做离断。损伤组：离断双侧生殖股神经生殖支及海绵体神经,断端以丝线结扎防止再生。神经移位组：离断双侧生殖股神经生殖支及海绵体神经后,双侧生殖股神经生殖支近端与海绵体神经远端行端端吻合。所有大鼠分别于术后4、8、12周行交配试验,观察并记录大鼠的交配行为。术后12周,各组大鼠在荧光金注射一周后分别行海绵体内压测定,电刺激相应神经时,监测并记录每只大鼠阴茎海绵体内压变化及动脉血压。结果：术后12周,假手术组和神经移位组分别可观察到8只和7只大鼠有“插入”性行为；而损伤组中仅1只大鼠有“插入”性行为,明显低于假手术组和神经移位组(P0.05)。电刺激神经移位组大鼠生殖股神经生殖支(供体神经),可见海绵体压显著升高,海绵体内压升高均值(AICP)为32.81±10.8mmHg,可达假手术组大鼠(62.11±7.67mmHg)的53%；而电刺激损伤组大鼠离断的海绵体神经近端,几乎未见海绵体内压升高,测得AICP为5.41±2.02mmHg,明显低于神经移位组和假手术组(P0.05)。假手术组、损伤组和神经移位组大鼠海绵体压升高的均值与平均动脉血压比值(△ICP/MAP)分别为：0.63±0.08,0.05±0.02和0.32±0.10,其中神经移位组和假手术组测得的比值显著高于损伤组(P0.05)。结论：生殖股神经生殖支移位海绵体神经后,部分大鼠可恢复自主交配行为,电刺激大鼠生殖股神经生殖支可引起海绵体内压显著升高,表明神经移位可部分修复大鼠的勃起功能。目的：探讨生殖股神经生殖支移位海绵体神经后阴茎海绵体NOS的表达以及组织形态学的改变。方法：将30只成年雄性Sprague-Dawley大鼠(约250-300g)随机分为三组,每组10只：假手术组,仅显露双侧生殖股神经及海绵体神经,不做离断。损伤组：离断双侧生殖股神经生殖支及海绵体神经,断端以丝线结扎防止再生。神经移位组：离断双侧生殖股神经生殖支及海绵体神经后,双侧生殖股神经生殖支近端与海绵体神经远端行端端吻合。术后12周,行功能学评估后,分别截取三组大鼠的阴茎海绵体组织,并分为两段,其中一段采用NADPH染色,于光学显微镜下观察并计数大鼠每侧阴茎背神经中蓝染的NOS阳性神经纤维。另一段则用于Masson染色以了解各组大鼠阴茎海绵体组织形态学的改变。结果：术后12周,神经移位组大鼠每侧阴茎背神经中均可见大量NOS阳性神经纤维,平均数量为58.67±13.3,明显高于损伤组(15.53±7.0)(P0.05),但低于假手术组(128.02±18.1)(P0.05)。阴茎海绵体Masson染色可见红染的为平滑肌细胞,蓝染的为胶原成分；三组大鼠阴茎海绵体切片内平滑肌和胶原的面积比分别为0.106±0.015,0.048±0.008和0.086±0.013,其中假手术组和神经移位组的比值显著高于损伤组(P0.05)。结论：生殖股神经生殖支移位海绵体神经后,大量NOS阳性的再生神经纤维重支配阴茎海绵体,可以明显减轻海绵体组织的纤维化。
[Abstract]:Objective: To investigate the morphological characteristics of the regenerated nerve after the transposition of the generative branch of the genital nerve to the cavernous nerve and the formation of the new erection reflex passage.
Methods: 30 adult male Sprague-Dawley rats (about 250-300g) were randomly divided into three groups, 10 in each group. The sham operation group was only exposed to the bilateral reproductive and cavernous nerve, and did not break off. The injured group was separated from the bilateral reproductive and cavernous nerves, and the broken ends were ligated with silk thread to prevent regeneration. After the lateral reproductive branch of the femoral nerve and the cavernous nerve, the proximal end of the bilateral reproductive branch of the reproductive nerve anastomosed with the distal end of the cavernous nerve. 12 weeks after the operation, the fluorescent gold solution (Fluorogold, FG) was injected into the right penis foot of the rats in each group, and the distal cavernous nerve was sectioning in each group. The nerve regeneration was evaluated by the myelinated nerve fiber count and electron microscopy, in which the anastomotic segment of the nerve in the nerve translocation group was further stained with the longitudinal section of the nerve to understand the continuity of the anastomotic segment of the nerve.
Results: the retrograde nerve tracer found that only a few FG positive neurons in the pelvic ganglion of the injured group and the nerve transfer group were only 6.3 + 3.4 and 5.4 + 2.9, respectively, far less than the sham operation rats (106.7 + 15.9), and the FG positive neurons were not found in the sham operation group and the injured group in the sections of the spinal cord. There were L1 and FG positive neurons in the spinal cord of the nerve translocation group, and the average number of FG positive neurons in the anterior horn of L2 was 2.7 + 1.0,5.3 + 1.2. nerve transect semi thin section of toluidine blue staining: the regenerated nerve of the cavernous nerve and the nerve transfer group of the sham operation group were all rich in blue stained myelinated nerve fibers. The number of myelinated nerve fibers in the distal cavernous nerve was significantly higher than that of 346.8 + 17.66 and 621.87 + 81.25 (16.68 + 6.08) (P0.05). Ultrastructural electron microscopy further observed that there were a large number of myelinated and unmyelinated nerve fibers in the sham operation group and the nerve translocation group, and almost no myelin sheath was found in the degeneration and necrosis, but the myelin sheath was widely changed in the injured group. In the nerve translocation group, the HE staining of the anastomotic segment of the rat showed that the genital branch of the genital nerve could grow into the cavernous nerve through the anastomotic stoma.
Conclusion: the reproductive branch of the reproductive femoral nerve translocation the cavernous nerve, the reproductive branch of the reproductive femoral nerve can grow into the corpus cavernosum, and establish a new erectile reflex efferent nerve pathway (spinal cord L1, L2 anterior horn motoneuron - reproductive nerve reproductive branch - regenerative nerve to penis corpus cavernous body).
Objective: To evaluate the recovery of erectile function after transposition of the genital nerve of reproductive system in rats.
Methods: 30 adult male Sprague-Dawley rats (about 250-300g) were randomly divided into three groups, 10 rats in each group: sham operation group, only bilateral reproductive femoral nerve and cavernous nerve were exposed. The injured group was separated from bilateral reproductive and cavernous nerve, and the broken ends were ligated with silk thread to prevent regeneration. The reproductive branches of the lateral reproductive and the cavernous nerve were anastomosed to the end end of the bilateral reproductive branch of the reproductive branch and the distal cavernous nerve. All rats were tested and recorded the mating behavior at 4,8,12 weeks after the operation. 12 weeks after the operation, the rats in each group were measured by the pressure of the sponge in the body after a week of injection of fluorescent gold. During the stimulation of the corresponding nerve, the pressure changes and arterial blood pressure in the corpus cavernosum of each rat were monitored and recorded.
Results: 12 weeks after operation, 8 and 7 rats in the sham operation group and the nerve transfer group were observed to have "insertion" behavior, while only 1 rats in the injured group had "insertion" behavior, obviously lower than the sham operation group and the nerve translocation group (P0.05). The mean pressure of body pressure increased significantly (AICP) was 32.81 + 10.8mmHg, which could reach 53% of the sham operation group (62.11 + 7.67mmHg), while the electrical stimulation injury group had almost no elevation of the cavernous nerve in the broken cavernous nerve, and the AICP was 5.41 + 2.02mmHg, obviously lower than that of the nerve translocation group and the sham operation group (P0.05). The mean and mean arterial pressure ratio (delta ICP/MAP) was 0.63 + 0.08,0.05 + 0.02 and 0.32 + 0.10 in the injury group and the nerve translocation group, respectively. The ratio between the nerve shift group and the sham operation group was significantly higher than that of the injury group (P0.05).
Conclusion: after the reproduction of the reproductive branch of the reproductive femoral nerve translocation the cavernous nerve, some rats can recover the spontaneous mating behavior. The reproductive branch of the reproductive femoral nerve can cause a significant increase in the internal pressure of the cavernous body, indicating that the nerve transfer can partially repair the erectile function of the rat.
Objective: To investigate the expression and histomorphology of NOS in corpus cavernosum after transferring the genital nerve to the cavernous nerve.
Methods: 30 adult male Sprague-Dawley rats (about 250-300g) were randomly divided into three groups, 10 rats in each group: sham operation group, only bilateral reproductive femoral nerve and cavernous nerve were exposed. The injured group was separated from bilateral reproductive and cavernous nerve, and the broken ends were ligated with silk thread to prevent regeneration. After 12 weeks of functional evaluation, the penis corpus cavernosum tissue of three groups of rats were divided and divided into two segments after 12 weeks of functional evaluation, and one of them was stained with the optical microscope and counted each rat under the optical microscope. The NOS positive nerve fibers in the dorsal penile nerve of the lateral penis were used. The other section was used for Masson staining to understand the histomorphological changes of corpus cavernosum in each group.
Results: 12 weeks after the operation, a large number of NOS positive nerve fibers in the dorsal nerve of the penis were found in the nerve translocation group, the average number was 58.67 + 13.3, which was significantly higher than that in the injury group (15.53 + 7) (P0.05), but lower than the sham operation group (128.02 + 18.1) (P0.05). The Masson staining of the penis corpus cavernosum showed that the red dye was the smooth muscle cells and the blue dye was collagen. The area ratio of smooth muscle and collagen in the cavernosum section of three groups of rats was 0.106 + 0.015,0.048 + 0.008 and 0.086 + 0.013 respectively, and the ratio of the sham operation group and the nerve translocation group was significantly higher than that of the injury group (P0.05).
Conclusion: a large number of NOS positive regenerative nerve fibers redominate the corpus cavernosum of the cavernous nerve after the reproduction of the reproductive branch of the reproductive femoral nerve, which can obviously reduce the fibrosis of the cavernous tissue.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R699.8

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