出生后不同年龄大鼠三叉神经节中伤害性感觉神经元的变化研究

发布时间：2018-03-19 03:08

本文选题：伤害性感觉神经元　切入点：酪氨酸激酶A(trkA)　出处：《深圳大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:机体的感觉神经节主要包括三叉神经节(trigeminal ganglion,TRG)和背根神经节(dorsal root ganglion,DRG),均由多种不同类型的感觉神经元组成,感觉神经元将特定的刺激转化成动作电位或分级电位,将感觉信息传送到脑或脊髓,其中包括了负责疼痛感受的伤害性感觉神经元,它们传递组织损伤或应对威胁的信号,是机体实现自我保护的重要途径。迄今为止,文献中缺少关于不同发育年龄TRG中伤害性感觉神经元组分及变化规律的报道,因此本实验旨在探讨大鼠TRG中伤害性感觉神经元在不同发育年龄的变化及其规律,并与大鼠背根神经节DRG中伤害性感觉神经元构成情况进行定量比较分析。本实验能够填补科研文献上的资料空缺,为疼痛及相关领域研究提供依据和参考。方法:选取不同发育年龄阶段的Sprague-Dawley(SD)大鼠,每个年龄组(出生后第0,3,7,14,21,60天)6只,未成年大鼠经麻醉后处死,成年大鼠则在局麻后进行心脏灌流,然后取其右侧的TRGs,成年大鼠同时从背部取同侧腰4、5、6节段(L4、L5、L6)的DRGs,所有组织立即分别浸泡于4%多聚甲醛中进行固定,经进一步脱水、透明和浸蜡后,制备成石蜡切片。用抗酪氨酸激酶A抗体(anti-tyrosine kinase Aantibody,trkA-Ab)和植物凝集素B4(Isolectin-B4,IB4)对切片中所有伤害性感觉神经元进行免疫荧光染色,并计算各类阳性标记神经元占神经节中全部感觉神经元的比率,定量数据均用均数±标准误进行表述,并进行统计学分析,p值小于0.05代表差异具有显著性。结果:从大鼠出生当天至成年,三叉神经节中表达trkA的神经元占整体神经元比率分别为 69.51±2.02%、60.15±1.38%、53.87±2.16%、35.1±1.40%、34.22±1.67%和31.85±0.75%;IB4染色阳性的神经元比率分别为17.77±0.70%、33.22±2.92%、41.36±5.84%、41.38±3.27%、43.83±5.06%和 38.22±4.10%;全部伤害性感觉神经元(表达trkA或被IB4阳性染色细胞)的比例依次为78.65±2.03%、88.90±2.58%、91.11±6.39%、69.10±3.52%、70.54±5.34%、54.71±3.49%;trkA与IB4结合蛋白共表达的神经元为数较少,分别为8.63±1.01%、8.93±1.17%、4.12±0.89%、7.42±1.24%、7.51±1.23%和15.36±0.96%。成年大鼠的背根神经节中,trkA阳性、IB4阳性、trkA或IB4单阳性,以及trkA和IB4双阳性的比率依次为 30.24±0.27%、39.58±1.03%、54.70±0.87%和 14.60±0.14%。结论:三叉神经节中,不介导神经性疼痛的trkA-Ab阳性神经元在生后发育的过程中逐渐减少,介导神经性疼痛的IB4阳性神经元则逐渐增多,这可能会引起神经性疼痛随年龄增长而增多;伤害性感觉神经元的总数量随着年龄的增长而减少,可能是疼痛敏感性降低的原因。上述各组数据在L4-L6 DRGs中保持一致,亦与成年大鼠TRGs中的数据相近,说明机体内有复杂的机制调控着发育进程不同的DRG和TRG伤害性感觉神经元并使它们的比例维持一致。
[Abstract]:Objective: the sensory ganglion consists of trigeminal ganglia trigeminal ganglion TRG and dorsal root ganglion DRGG, all of which are composed of various types of sensory neurons. Sensory neurons transform specific stimuli into action potentials or graded potentials. Sending sensory information to the brain or spinal cord, including nociceptive sensory neurons responsible for pain perception, which signals tissue damage or response to threats, is an important way for the body to protect itself. There is a lack of reports on the composition and changes of nociceptive sensory neurons in TRG at different developmental ages. Therefore, the purpose of this study was to investigate the changes and regularity of nociceptive sensory neurons in TRG of rats at different developmental ages. The results were compared with those of the nociceptive sensory neurons in rat dorsal root ganglion (DRG). Methods: Sprague-Dawley SD rats of different developmental ages were selected. Adult rats were perfused with the heart after local anesthesia, and then TRGs were taken from the right side of the rat. The DRGs of the adult rats were simultaneously taken from the same side of the back of the same side. All the tissues were immediately immersed in 4% paraformaldehyde for further dehydration. After transparent and wax immersion, paraffin sections were prepared. All nociceptive sensory neurons were stained with anti-tyrosine kinase AantibodytrkA-Aband phytoagglutinin B4 Isolectin-B4IB4. The ratio of all kinds of positive labeled neurons to all sensory neurons in ganglion was calculated, and the quantitative data were misrepresented by mean 卤standard. Results: from the day of birth to adulthood, there was significant difference between the two groups. The percentage of neurons expressing trkA in trigeminal ganglion was 69.51 卤2.02and 60.15 卤1.38and 53.87 卤2.163.87 卤1.400.35.1 卤1.400.75 卤34.22 卤1.67% and 31.85 卤0.7575 respectively. The positive rate of IB4 staining in trigeminal ganglion was 17.77 卤0.703.22 卤2.9221.36 卤5.841.36 卤5.8441.38 卤3.27 卤43.83 卤5.06% and 38.22 卤4.10.The percentage of all nociceptive sensory neurons (expressing trkA or IB4 positive cells) was 17.77 卤0.703.38 卤3.27 卤43.83 卤5.06% and 38.22 卤4.10respectively. The ratio was 78.65 卤2.03, 88.90 卤2.58 and 91.11 卤6.39. The number of neurons co-expressing IB4 binding protein was less than that of 70.54 卤5.34 卤54.71 卤3.49trkA and 69.10 卤3.52, respectively, and the ratio of 78.65 卤2.03 and 88.90 卤2.58 was 91.11 卤6.39 and 79.10 卤3.52 and 54.71 卤3.49trkA respectively. 8.63 卤1.01T = 8.93 卤1.171.17 卤4.12 卤0.89 and 7.42 卤1.24, 7.51 卤1.23% and 15.36 卤0.96.The positive rates of trkA and IB4 in dorsal root ganglion of adult rats were 30.24 卤0.2739.58 卤1.039.58 卤1.034.70 卤0.87% and 14.60 卤0.140.Conclusion: in the trigeminal ganglion, the positive rate of TRA and IB4 were 30.24 卤0.2739.58 卤1.030.70 卤0.87% and 14.60 卤0.14.Conclusion: in the trigeminal ganglion, the positive rate of trkA and IB4 is 30.24 卤0.2739.58 卤1.039.58 卤1.034.70 卤0.87% and 14.60 卤0.14.4 respectively. The number of trkA-Ab positive neurons that did not mediate neuropathic pain decreased gradually during postnatal development, while the IB4 positive neurons involved in neuropathic pain increased gradually, which may cause neuropathic pain to increase with age. The total number of nociceptive sensory neurons decreased with age, which may be the reason for the decrease in pain sensitivity. These data were consistent in L4-L6 DRGs and similar to those in adult rat TRGs. It is suggested that there are complex mechanisms in the body to regulate the nociceptive neurons of DRG and TRG in different developmental processes and to maintain a consistent proportion of them.
【学位授予单位】：深圳大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R338

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