新劑型神經(jīng)生長因子聯(lián)合丙戊酸促進(jìn)大鼠坐骨神經(jīng)損傷再生協(xié)同作用的實驗研究
發(fā)布時間：2018-11-12 18:54
【摘要】：周圍神經(jīng)損傷實質(zhì)上是神經(jīng)元軸突的損傷，周圍神經(jīng)損傷后軸突逆向運輸提供的神經(jīng)營養(yǎng)因子驟減，導(dǎo)致部分神經(jīng)元死亡，再生乏力。研究表明，靶組織合成的神經(jīng)營養(yǎng)物質(zhì)有利于損傷神經(jīng)的再生與修復(fù)。神神經(jīng)生長因子在神經(jīng)損傷的再生過程中起著極其重要的營養(yǎng)和調(diào)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(dān)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(yīng)用有促進(jìn)PC12細(xì)胞分化的協(xié)同作用的基礎(chǔ)上，采用Wistar大鼠坐骨神經(jīng)損傷后創(chuàng)建硅膠管神經(jīng)再生室的動物模型，將新劑型NGF(注射入神經(jīng)再生室內(nèi))與VPA（口服）聯(lián)合使用，觀察神經(jīng)損傷后軸突再生的數(shù)量、質(zhì)量及神經(jīng)電生理功能恢復(fù)變化情況。 實驗結(jié)果顯示：1、B、C、D、E、F、G各組在D42時間點坐骨神經(jīng)的肌電圖檢測呈現(xiàn)恢復(fù)期表現(xiàn)，通過神經(jīng)再生室遠(yuǎn)端所產(chǎn)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(jīng)過神經(jīng)再生室后均有明顯AMP衰減，同時經(jīng)過神經(jīng)再生室的CV均有明顯減慢，各組與A相比，均有統(tǒng)計學(xué)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(xué)意義（P0.05），，B、C兩組之間相差不大無統(tǒng)計學(xué)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(xué)差異（P0.05）。2、觀察各組再生神經(jīng)軸突總面積及相對恢復(fù)率結(jié)果顯示各組再生的神經(jīng)均通過吻合口：A組神經(jīng)軸突的髓鞘較厚，數(shù)目較多，平均直徑大，其余各組光鏡下觀察見間質(zhì)增多，再生的軸突髓鞘細(xì)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(fù)率組間也有差異，與A組對比有明顯統(tǒng)計學(xué)差異（P0.05）各組之間進(jìn)行比較分析，其中E、F、G三組軸突總面積及軸突面積恢復(fù)率組間無統(tǒng)計學(xué)差異（P0.05），E、F、G三組軸突總面積及軸突面積恢復(fù)率分別均高于B、C、D三組具有統(tǒng)計學(xué)差異（P0.05）；D組軸突總面積及軸突面積恢復(fù)率高于B、C兩組具有統(tǒng)計學(xué)差異（P0.05）；B、C兩組軸突總面積及軸突面積恢復(fù)率組間相比無統(tǒng)計學(xué)差異（P0.05）。 研究結(jié)論：硅膠管神經(jīng)再生室內(nèi)使用新劑型NGF可以促進(jìn)神經(jīng)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(jīng)再生室模型可以有效地進(jìn)行促進(jìn)神經(jīng)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(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R651.3
本文編號：2327924
[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(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R651.3
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本文編號：2327924
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