背根节慢性压迫模型小细胞交感敏化的NO-cGMP-PKG信号通路研究
发布时间：2018-11-02 19:07
【摘要】：交感维持性痛（SMP）是临床一种常见的慢性痛，SMP是指通过阻滞支配疼痛发生区域的交感神经而得到缓解的疼痛，这类疼痛常表现出依赖交感神经活动及循环中儿茶酚胺水平的特征，即交感敏化。交感敏化在细胞水平的表现为交感神经刺激或肾上腺素类物质（NE）加入后，可引起神经元兴奋性异常增高，表现为产生放电所需刺激强度的降低或原有异位自发放电的增多。DRG内的小细胞是传递痛觉信息的神经元。在神经病理痛过程中，这类神经元存在超兴奋现象，有报道C纤维上存在交感敏化，然而其胞体上是否存在交感敏化及参与调节其交感敏化的细胞内信号机制迄今未见报道。本研究拟回答如下问题：（一）在背根节慢性压迫（CCD）模型背根节小细胞上是否存在交感敏化现象？（二）CCD模型后小细胞上如果存在交感敏化现象，参与介导敏化的胞内信号通路是哪条？本实验按照这一思路进行研究。 目的：揭示CCD模型背根节小细胞上是否存在交感敏化现象及其产生的信号通路。 方法：本研究以慢性背根节压迫模型小鼠和正常C57小鼠为实验对象，应用行为学检测技术、膜片钳技术，观察损伤的DRG小细胞上的交感敏化现象，在此基础上利用各种药物探讨交感敏化的细胞内信号转导机制。 结果： 1、 CCD建模后，模型组出现痛觉过敏现象。 缩足阈值（PWT）开始下降，7d时明显低于对照组。CCD组与造模前相比，造模后7dPTW下降至0.13g，而对照组动物的PWT则一直较恒定在0.32g。 2、 CCD建模后小细胞出现交感敏化现象。 在CCD后DRG上直径不超过25μm的神经元上，应用NE的反应有单纯兴奋作用、单纯抑制作用、无作用和先兴奋后抑制的作用。90例神经元中，单纯兴奋58例（64%），单纯抑制14例（16%），无作用16例（18%），先兴奋后抑制2例（2%）。而正常对照组10例神经元中，单纯兴奋2例，单纯抑制2例，无作用6例。 3、 α2受体阻断剂可以抑制交感敏化现象的产生 在CCD组出现交感敏化的小细胞上，通过利用α2受体阻断剂育亨宾（Yohimbine）（10μM），可以抑制交感敏化现象的产生（n=8），据此推论在交感敏化现象中去甲肾上腺素作用受体为α2受体。 4、 NO-cGMP-PKG信号通路参与介导NE对小细胞的兴奋的作用。 通过利用NO供体SNAP（100μM），神经元的放电个数由(8.36±1.08)增加至(11.72±1.26),(P0.05)；cGMP阻断剂ODQ则抑制了NE的兴奋效应,放电个数由(14.09±2.02)减少至(8.64±1.88),(P0.05)；PKG阻断剂KT5823同样抑制了NE的兴奋效应,放电个数由(15.56±2.07)减少到(9±0.75),(P0.05)。 结论： 1、 CCD后部分DRG小细胞出现交感敏化现象。 2、交感敏化现象中去甲肾上腺素作用受体为α2受体。 3、NO-cGMP-PKG信号通路参与了交感敏化现象的发生。大量文献报道，受损DRG神经元的异位自发放电与神经病理性痛行为密切相关，关于异位自发放电的通道电流机制也有大量研究，研究结果显示多种通道电流大小的改变是神经病理痛发生的一个重要因素。但迄今为止，还没有文献报道某种特定的放电模式（如周期放电、burst等）有着与其对应的疼痛特征，如外周神经损伤后的痛觉过敏、触诱发痛和自发痛等，因此，很难将通道电流的变化与痛信号模式的关系作出准确判断，影响了痛信号产生机制的深入研究。 目的：本论文的第二部分实验拟在证明“诱发簇放电（Evoked bursting, EB）”可能是触诱发痛的痛信号的基础上，对EB现象及其它神经病理性痛的通道电流机制进行深一步的研究。方法：以正常大鼠和慢性背根节压迫大鼠为实验对象，利用离体整节DRG标本及膜片钳技术，对EB及可能的交感敏化的离子通道机制进行深入探讨。 结果： α-DTX敏感钾电流（IDTX）参与介导诱发簇放电（EB）发生。 α-DTX是低阈值K+通道的阻断剂，对Kv1.1、1.2高度敏感。在CCD后的DRG大神经元上应用1nM的α-DTX使得神经元兴奋性显著增加，EB的串长增加（n=5），部分神经元还出现自发放电活动（n＝2）。 结论： 在CCD模型中，IDTX参与介导DRG大神经元诱发簇放电（EB）的发生。
[Abstract]:An SMP is a common chronic pain in which the SMP means pain relieved by blocking the sympathetic nerve in the area where pain occurs, which often represents a characteristic that depends on the level of the sympathetic nerve activity and circulation in the circulation, i.e. sensitisation. After the addition of sympathetic stimulation or epinephrine (NE) to the cell level, the increase of the excitability of the neuron may be caused by the decrease of the stimulation intensity or the increase of the original ectopic spontaneous discharge. Small cells in DRG are neurons that deliver pain information. In the course of neuropathic pain, there are hyperstimulation phenomena in this kind of neuron, and there is a sensitization on C-fiber. However, there is no report on whether there is photosensitization on its body and the intracellular signal mechanism involved in the regulation of its photosensitization. This study is intended to answer the following questions: (1) Is there any sensitization on small cells of dorsal root ganglion of dorsal root ganglion chronic compression (CCD) model? (2) What is the intracellular signaling pathway involved in mediating sensitization on small cells after CCD model? This experiment was conducted according to this idea. Objective: To reveal the presence or absence of dye sensitization and the signal produced by CCD model dorsal root ganglion cells. Methods: In this study, chronic dorsal root ganglion compression model mice and normal C57 mice were used as experimental subjects, and behavioral detection technique, TUNEL technique were used to observe the delivery of injured DRG cells. Sensitization phenomenon, on the basis of this, the use of various drugs to explore the intracellular signaling of dye-sensitized cells signal transduction Mechanism. Result: 1. After the CCD is modeled, There was hyperalgesia in the model group. The foot threshold (PWT) was open. Compared with the control group, 7dPTW decreased to 0. 13g after modeling and P in the control group compared with the control group. wt has always been constant at 0. 32g. 2, Photosensitization of small cells after CCD modeling was observed. On the DRG with a diameter of no more than 25. m u.m, the action of NE was applied to the neurons with pure excitement, simple inhibition, no action and inhibition after first excitation. In 90 cases of neurons, pure excitement was 5. 8 cases (64%), simple inhibition of 14 cases (16%), no effect 1 In 6 cases (18%), 2 cases (2%) were inhibited by first excitation. 2 cases were simply excited, only 2 cases were inhibited, and there was no action in 6 cases. 3. The Cd2 receptor blocker can inhibit the generation of dye-sensitization phenomenon on small cells sensitized by photosensitization in the CCD group, and Yuhenbin (Yohimbine) can be obtained by using the B2C 2 receptor blocker. (10 & mu; M), the generation of the dye sensitization phenomenon can be suppressed (n = 8), The result of this inference is that the receptor of noradrenalin plays a role in the photosensitization phenomenon. 4, NO. cGMP-PKG signaling pathway plays an important role in mediating NE's stimulation of small cells. By using NO donor SNAP (100. mu.M), the number of discharges of neurons is increased from (8.36-1.08) to (11.72-1.26), (P0.05); cGMP blockers ODQ inhibit the excitation effect of NE and the number of discharges is (1). 4. 09 (2.02) reduced to (8.64, 1.88), (P0.05); PKG blocker KT5823 also inhibited NE's excitation effect and the number of discharges From (1 5.56 (2.07) reduced to (9) 0. 75), (p0.05). Conclusion: 1. CCD rear part D in RG small cell, there was a sensitization phenomenon. The receptor of gondolin is two receptors. 3, NO-cGMP-PKG signaling pathway is involved in the occurrence of sensitization. The literature reports that ectopic spontaneous discharge of damaged DRG neurons is closely related to neuropathic pain behavior, and it is related to ectopic spontaneous discharge. There is also a large number of studies in the channel current mechanism, and the results show that the change of the current size of multiple channels is an important factor in the occurrence of neuropathic pain. However, no specific discharge pattern (such as periodic discharge, b) has been reported to date. urst et al.) have pain characteristics corresponding thereto, such as hyperalgesia after peripheral nerve injury, touch-induced pain and spontaneous pain, etc., Therefore, it is difficult to change the change of the channel current and the pain signal. The relationship between patterns is accurately judged, which affects the deep research of the mechanism of pain signal generation. Objective: The second part of this paper is to prove that "induced cluster discharge" (EB) On the basis of the pain signal which may be the touch-induced pain, the mechanism of the channel current of EB and other neuropathic pain was studied in this paper. The method: The rats were pressed by normal rats and chronic dorsal root ganglion. experimental pair The EB and possible photosensitization were sensitised by the use of an off-body DRIG specimen, as well as a nuclear fission technique. The ion channel mechanism is discussed in detail. The results show that the K-DTX sensitive potassium current (IDTX) is involved in mediating induced cluster discharge (EB), which is a low-threshold K + channel blocker which is highly sensitive to Kv1.1, 1,2 highly. On the DRG neurons following CCD Application 1 nM OPG-DTX resulted in a significant increase in neuronal excitability, with an increase in the number of Bs (n = 5), in part
【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R-332
本文编号：2306694
[Abstract]:An SMP is a common chronic pain in which the SMP means pain relieved by blocking the sympathetic nerve in the area where pain occurs, which often represents a characteristic that depends on the level of the sympathetic nerve activity and circulation in the circulation, i.e. sensitisation. After the addition of sympathetic stimulation or epinephrine (NE) to the cell level, the increase of the excitability of the neuron may be caused by the decrease of the stimulation intensity or the increase of the original ectopic spontaneous discharge. Small cells in DRG are neurons that deliver pain information. In the course of neuropathic pain, there are hyperstimulation phenomena in this kind of neuron, and there is a sensitization on C-fiber. However, there is no report on whether there is photosensitization on its body and the intracellular signal mechanism involved in the regulation of its photosensitization. This study is intended to answer the following questions: (1) Is there any sensitization on small cells of dorsal root ganglion of dorsal root ganglion chronic compression (CCD) model? (2) What is the intracellular signaling pathway involved in mediating sensitization on small cells after CCD model? This experiment was conducted according to this idea. Objective: To reveal the presence or absence of dye sensitization and the signal produced by CCD model dorsal root ganglion cells. Methods: In this study, chronic dorsal root ganglion compression model mice and normal C57 mice were used as experimental subjects, and behavioral detection technique, TUNEL technique were used to observe the delivery of injured DRG cells. Sensitization phenomenon, on the basis of this, the use of various drugs to explore the intracellular signaling of dye-sensitized cells signal transduction Mechanism. Result: 1. After the CCD is modeled, There was hyperalgesia in the model group. The foot threshold (PWT) was open. Compared with the control group, 7dPTW decreased to 0. 13g after modeling and P in the control group compared with the control group. wt has always been constant at 0. 32g. 2, Photosensitization of small cells after CCD modeling was observed. On the DRG with a diameter of no more than 25. m u.m, the action of NE was applied to the neurons with pure excitement, simple inhibition, no action and inhibition after first excitation. In 90 cases of neurons, pure excitement was 5. 8 cases (64%), simple inhibition of 14 cases (16%), no effect 1 In 6 cases (18%), 2 cases (2%) were inhibited by first excitation. 2 cases were simply excited, only 2 cases were inhibited, and there was no action in 6 cases. 3. The Cd2 receptor blocker can inhibit the generation of dye-sensitization phenomenon on small cells sensitized by photosensitization in the CCD group, and Yuhenbin (Yohimbine) can be obtained by using the B2C 2 receptor blocker. (10 & mu; M), the generation of the dye sensitization phenomenon can be suppressed (n = 8), The result of this inference is that the receptor of noradrenalin plays a role in the photosensitization phenomenon. 4, NO. cGMP-PKG signaling pathway plays an important role in mediating NE's stimulation of small cells. By using NO donor SNAP (100. mu.M), the number of discharges of neurons is increased from (8.36-1.08) to (11.72-1.26), (P0.05); cGMP blockers ODQ inhibit the excitation effect of NE and the number of discharges is (1). 4. 09 (2.02) reduced to (8.64, 1.88), (P0.05); PKG blocker KT5823 also inhibited NE's excitation effect and the number of discharges From (1 5.56 (2.07) reduced to (9) 0. 75), (p0.05). Conclusion: 1. CCD rear part D in RG small cell, there was a sensitization phenomenon. The receptor of gondolin is two receptors. 3, NO-cGMP-PKG signaling pathway is involved in the occurrence of sensitization. The literature reports that ectopic spontaneous discharge of damaged DRG neurons is closely related to neuropathic pain behavior, and it is related to ectopic spontaneous discharge. There is also a large number of studies in the channel current mechanism, and the results show that the change of the current size of multiple channels is an important factor in the occurrence of neuropathic pain. However, no specific discharge pattern (such as periodic discharge, b) has been reported to date. urst et al.) have pain characteristics corresponding thereto, such as hyperalgesia after peripheral nerve injury, touch-induced pain and spontaneous pain, etc., Therefore, it is difficult to change the change of the channel current and the pain signal. The relationship between patterns is accurately judged, which affects the deep research of the mechanism of pain signal generation. Objective: The second part of this paper is to prove that "induced cluster discharge" (EB) On the basis of the pain signal which may be the touch-induced pain, the mechanism of the channel current of EB and other neuropathic pain was studied in this paper. The method: The rats were pressed by normal rats and chronic dorsal root ganglion. experimental pair The EB and possible photosensitization were sensitised by the use of an off-body DRIG specimen, as well as a nuclear fission technique. The ion channel mechanism is discussed in detail. The results show that the K-DTX sensitive potassium current (IDTX) is involved in mediating induced cluster discharge (EB), which is a low-threshold K + channel blocker which is highly sensitive to Kv1.1, 1,2 highly. On the DRG neurons following CCD Application 1 nM OPG-DTX resulted in a significant increase in neuronal excitability, with an increase in the number of Bs (n = 5), in part
【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R-332
【参考文献】
相关期刊论文 前2条
1 ;Involvement of hyperpolarization-activated,cyclic nucleotide-gated cation channels in dorsal root ganglion in neuropathic pain[J];生理学报;2008年05期
2 Judith A.Strong;;Recent evidence for activity-dependent initiation of sympathetic sprouting and neuropathic pain[J];生理学报;2008年05期
，本文编号：2306694
本文链接：https://www.wllwen.com/xiyixuelunwen/2306694.html
最近更新
教材专著
