發(fā)布時間：2018-11-12 18:54

【摘要】：周圍神經損傷實質上是神經元軸突的損傷，周圍神經損傷后軸突逆向運輸提供的神經營養(yǎng)因子驟減，導致部分神經元死亡，再生乏力。研究表明，靶組織合成的神經營養(yǎng)物質有利于損傷神經的再生與修復。神神經生長因子在神經損傷的再生過程中起著極其重要的營養(yǎng)和調控作用。但NGF存在無法通過血腦屏障、全身用藥毒副作用大、水溶液中生物活性半衰期短等缺陷，基于此我們利用聚乙二醇/聚（γ-乙基-L-谷氨酸酯）（PEG-PELG）水凝膠擔載NGF，制備出具有緩釋能力的新劑型NGF。在體外試驗證實NGF與VPA聯(lián)合應用有促進PC12細胞分化的協(xié)同作用的基礎上，采用Wistar大鼠坐骨神經損傷后創(chuàng)建硅膠管神經再生室的動物模型，將新劑型NGF(注射入神經再生室內)與VPA（口服）聯(lián)合使用，觀察神經損傷后軸突再生的數量、質量及神經電生理功能恢復變化情況。 實驗結果顯示：1、B、C、D、E、F、G各組在D42時間點坐骨神經的肌電圖檢測呈現(xiàn)恢復期表現(xiàn)，通過神經再生室遠端所產生的MUAP波形欠規(guī)則、離散，時限延長，各段波形不完全相同，經過神經再生室后均有明顯AMP衰減，同時經過神經再生室的CV均有明顯減慢，各組與A相比，均有統(tǒng)計學意義（P0.05）；其中D、E、F、G四組之間相比有差異，但無統(tǒng)計學意義（P0.05），，B、C兩組之間相差不大無統(tǒng)計學意義（P0.05），D、E、F、G四組與B、C兩組相比較AMP衰減率更小、CV增高明顯存在統(tǒng)計學差異（P0.05）。2、觀察各組再生神經軸突總面積及相對恢復率結果顯示各組再生的神經均通過吻合口：A組神經軸突的髓鞘較厚，數目較多，平均直徑大，其余各組光鏡下觀察見間質增多，再生的軸突髓鞘細小、分布分散，計算得出軸突總面積較空白組明顯減少，軸突面積恢復率組間也有差異，與A組對比有明顯統(tǒng)計學差異（P0.05）各組之間進行比較分析，其中E、F、G三組軸突總面積及軸突面積恢復率組間無統(tǒng)計學差異（P0.05），E、F、G三組軸突總面積及軸突面積恢復率分別均高于B、C、D三組具有統(tǒng)計學差異（P0.05）；D組軸突總面積及軸突面積恢復率高于B、C兩組具有統(tǒng)計學差異（P0.05）；B、C兩組軸突總面積及軸突面積恢復率組間相比無統(tǒng)計學差異（P0.05）。 研究結論：硅膠管神經再生室內使用新劑型NGF可以促進神經再生，且可以表現(xiàn)出高劑量效果優(yōu)于低劑量。低劑量新劑型NGF聯(lián)合使用VPA效果與高劑量及高劑量聯(lián)合VPA效果類似，具有協(xié)同作用。硅膠管神經再生室模型可以有效地進行促進神經再生藥物效果的測定。
[Abstract]:Peripheral nerve injury is essentially neuronal axonal injury. After peripheral nerve injury, the neurotrophic factor provided by reverse axon transport decreases sharply, resulting in the death of some neurons and the lack of regeneration. The results showed that the neurotrophic substances synthesized from target tissues were beneficial to the regeneration and repair of injured nerves. Nerve growth factor (NGF) plays an important role in the regeneration of nerve injury. However, NGF can not pass through the blood-brain barrier, the toxicity and side effects of systemic medication are large, and the bioactivity half-life in aqueous solution is short. Therefore, we use polyethylene glycol / poly (緯 -ethyl-L-glutamate) (PEG-PELG) hydrogel to support NGF,. Preparation of a new formulation NGF. with slow release ability On the basis of in vitro experiment to prove that NGF and VPA can promote the differentiation of PC12 cells, an animal model of silicone tube nerve regeneration chamber was established by using Wistar rats after sciatic nerve injury. A new dosage form, NGF (injected into nerve regeneration chamber) and VPA (oral administration), were used to observe the quantity, quality and electrophysiological function of axon regeneration after nerve injury. The results showed that: 1 the electromyogram (EMG) of sciatic nerve in D42 time group showed convalescence, and the MUAP waveform produced at the distal end of nerve regeneration chamber was irregular, dispersed and prolonged. The waveforms of each segment were not exactly the same, there was obvious AMP attenuation after the nerve regeneration chamber, and the CV of the nerve regeneration chamber was significantly slower than that of the A group (P0.05). There was significant difference among the four groups, but there was no significant difference between the four groups (P0.05), but there was no significant difference between the two groups (P0.05), while there was no significant difference between the two groups (P0.05). The attenuation rate of AMP in group C was smaller than that in group C, and there was a significant difference in the increase of CV between group C and group C (P0.05). The total area and relative recovery rate of regenerated axons in each group were observed. The results showed that the regenerated nerves in each group passed through the anastomotic stoma: the myelin sheath of nerve axons in group A was thicker, the number of axons was larger, the average diameter was larger, and the interstitial mass was increased under light microscope in other groups. The regenerated axonal myelin sheath was small and scattered. The total area of axon was significantly reduced and the recovery rate of axon area was also different between the two groups. There was significant statistical difference between group A and group A (P0.05). There was no significant difference in the total area of axon and the recovery rate of axon area among the three groups (P0.05). The total area of axon and the recovery rate of the area of axon in the three groups were higher than those in the group B (P 0.05). Group D had statistical difference (P0.05); The total area of axon and the recovery rate of axon area in group D were higher than those in group B and C (P0.05), but there was no significant difference in total area of axon and recovery rate of axon area between group B and C (P0.05). Conclusion: the new dosage form of NGF can promote nerve regeneration, and the effect of high dose is better than that of low dose. The effect of low dose NGF combined with VPA is similar to that of high dose and high dose VPA, and has synergistic effect. The model of silicone tube nerve regeneration chamber can be used to determine the effect of drugs for promoting nerve regeneration.
【學位授予單位】：吉林大學
【學位級別】：博士
【學位授予年份】：2015
【分類號】：R651.3

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