水蛭促进大鼠坐骨神经损伤修复的分子机制的实验研究

发布时间：2018-01-19 14:24

本文关键词： 水蛭坐骨神经修复分子机制　出处：《南京中医药大学》2017年硕士论文　论文类型：学位论文

【摘要】：周围神经损伤是临床常见的创伤性疾病,尽管由于显微外科的飞速发展,神经修复后功能恢复有所改善,但实际疗效仍难以达到理想水平。为了更好、更快的促进神经的恢复与再生,广大学者对此开展了大量的基础研究和临床应用研究。目前临床使用较多的以西药为主,这些常用药物的疗效有限,并且价格昂贵,经济负担沉重。因此,发挥中医中药的优势,运用中西医结合手段,探求高效低毒的修复神经损伤的药物,并阐明其分子机理,对于临床治疗有积极的意义。本实验主要研究水蛭作用于受损周围神经修复的分子机制,尤其是对神经生长因子、脑源性神经影响因子、神经生长相关蛋白等相关基因表达的定性、定量研究。目的模拟大鼠坐骨神经损伤模型,通过水蛭制剂灌胃治疗后,观察大鼠神经恢复的情况,同时测定细胞凋亡及神经生长因子、脑源性神经影响因子、神经生长相关蛋白等相关基因的表达情况。方法选取雄性SD大鼠100只,制备大鼠左侧坐骨神经损伤模型,随机分为4组,即水蛭组、弥可保组、生理盐水阴性对照组和空白对照组。水蛭组、弥可保组、生理盐水组术后预防感染3天,第4天起分别予以水蛭水煎液、弥可保片水溶解液和生理盐水灌胃。术后第4、8周,在电镜下观察神经组织损伤部位的恢复情况,并采取TUNEL法检测L5背根神经节的细胞凋亡情况,同时应用免疫组化法检测NGF、BDNF、GAP-43的相关蛋白的表达。结果水蛭组坐骨神经组织恢复情况较好,并且其NGF、BDNF、GAP-43的表达分别优于对照组,而凋亡细胞的数量少于对照组,有统计学意义差异。结论水蛭可能通过促进NGF、BDNF、GAP-43等神经生长修复相关经生长修复相关因子的表达,抑制细胞凋亡,从而对大鼠坐骨神经损伤发挥保护作用。
[Abstract]:Peripheral nerve injury is a common traumatic disease in clinic. Although the functional recovery after nerve repair has been improved due to the rapid development of microsurgery, the actual curative effect is still difficult to reach the ideal level. More rapid promotion of nerve recovery and regeneration, the majority of scholars have carried out a large number of basic research and clinical application research. At present, most of the clinical use of Western medicine, these commonly used drugs have limited efficacy. And expensive, heavy economic burden. Therefore, give play to the advantages of traditional Chinese medicine, the use of integrated traditional Chinese and Western medicine, explore the high efficiency and low toxicity of nerve repair drugs, and clarify its molecular mechanism. This experiment mainly studies the molecular mechanism of leech acting on the repair of injured peripheral nerve, especially on nerve growth factor and brain-derived neurogenic factor. Objective to simulate the rat model of sciatic nerve injury and observe the recovery of nerve after oral administration of leech preparation. Apoptosis and nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), nerve growth associated protein (NGP) and other related genes were determined. Methods 100 male SD rats were selected. The model of left sciatic nerve injury in rats was established and randomly divided into 4 groups: leech group, mycopol group, normal saline negative control group and blank control group. The rats in the saline group were treated with leech decoction, water soluble solution and normal saline solution respectively from the 4th day after operation, and the rats in the saline group were given intragastric administration for 4 weeks after operation. The recovery of injured nerve tissue was observed under electron microscope, the apoptosis of L5 dorsal root ganglion was detected by TUNEL method, and the expression of BDNF in L5 dorsal root ganglion was detected by immunohistochemical method. Results the recovery of sciatic nerve tissue in Hirudo group was better than that in control group, and the expression of BDNF / GAP-43 in Hirudo group was higher than that in control group. The number of apoptotic cells was less than that of the control group, there was significant difference. Conclusion Leech may promote the expression of NGF, BDNF, GAP-43 and other nerve growth repair related factors. Inhibition of apoptosis, so as to play a protective role against sciatic nerve injury in rats.
【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

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