背根節(jié)慢性壓迫模型小細(xì)胞交感敏化的NO-cGMP-PKG信號(hào)通路研究
發(fā)布時(shí)間：2018-11-02 19:07
【摘要】：交感維持性痛（SMP）是臨床一種常見(jiàn)的慢性痛，SMP是指通過(guò)阻滯支配疼痛發(fā)生區(qū)域的交感神經(jīng)而得到緩解的疼痛，這類疼痛常表現(xiàn)出依賴交感神經(jīng)活動(dòng)及循環(huán)中兒茶酚胺水平的特征，即交感敏化。交感敏化在細(xì)胞水平的表現(xiàn)為交感神經(jīng)刺激或腎上腺素類物質(zhì)（NE）加入后，可引起神經(jīng)元興奮性異常增高，表現(xiàn)為產(chǎn)生放電所需刺激強(qiáng)度的降低或原有異位自發(fā)放電的增多。DRG內(nèi)的小細(xì)胞是傳遞痛覺(jué)信息的神經(jīng)元。在神經(jīng)病理痛過(guò)程中，這類神經(jīng)元存在超興奮現(xiàn)象，有報(bào)道C纖維上存在交感敏化，然而其胞體上是否存在交感敏化及參與調(diào)節(jié)其交感敏化的細(xì)胞內(nèi)信號(hào)機(jī)制迄今未見(jiàn)報(bào)道。本研究擬回答如下問(wèn)題：（一）在背根節(jié)慢性壓迫（CCD）模型背根節(jié)小細(xì)胞上是否存在交感敏化現(xiàn)象？（二）CCD模型后小細(xì)胞上如果存在交感敏化現(xiàn)象，參與介導(dǎo)敏化的胞內(nèi)信號(hào)通路是哪條？本實(shí)驗(yàn)按照這一思路進(jìn)行研究。 目的：揭示CCD模型背根節(jié)小細(xì)胞上是否存在交感敏化現(xiàn)象及其產(chǎn)生的信號(hào)通路。 方法：本研究以慢性背根節(jié)壓迫模型小鼠和正常C57小鼠為實(shí)驗(yàn)對(duì)象，應(yīng)用行為學(xué)檢測(cè)技術(shù)、膜片鉗技術(shù)，觀察損傷的DRG小細(xì)胞上的交感敏化現(xiàn)象，在此基礎(chǔ)上利用各種藥物探討交感敏化的細(xì)胞內(nèi)信號(hào)轉(zhuǎn)導(dǎo)機(jī)制。 結(jié)果： 1、 CCD建模后，模型組出現(xiàn)痛覺(jué)過(guò)敏現(xiàn)象。 縮足閾值（PWT）開(kāi)始下降，7d時(shí)明顯低于對(duì)照組。CCD組與造模前相比，造模后7dPTW下降至0.13g，而對(duì)照組動(dòng)物的PWT則一直較恒定在0.32g。 2、 CCD建模后小細(xì)胞出現(xiàn)交感敏化現(xiàn)象。 在CCD后DRG上直徑不超過(guò)25μm的神經(jīng)元上，應(yīng)用NE的反應(yīng)有單純興奮作用、單純抑制作用、無(wú)作用和先興奮后抑制的作用。90例神經(jīng)元中，單純興奮58例（64%），單純抑制14例（16%），無(wú)作用16例（18%），先興奮后抑制2例（2%）。而正常對(duì)照組10例神經(jīng)元中，單純興奮2例，單純抑制2例，無(wú)作用6例。 3、 α2受體阻斷劑可以抑制交感敏化現(xiàn)象的產(chǎn)生 在CCD組出現(xiàn)交感敏化的小細(xì)胞上，通過(guò)利用α2受體阻斷劑育亨賓（Yohimbine）（10μM），可以抑制交感敏化現(xiàn)象的產(chǎn)生（n=8），據(jù)此推論在交感敏化現(xiàn)象中去甲腎上腺素作用受體為α2受體。 4、 NO-cGMP-PKG信號(hào)通路參與介導(dǎo)NE對(duì)小細(xì)胞的興奮的作用。 通過(guò)利用NO供體SNAP（100μM），神經(jīng)元的放電個(gè)數(shù)由(8.36±1.08)增加至(11.72±1.26),(P0.05)；cGMP阻斷劑ODQ則抑制了NE的興奮效應(yīng),放電個(gè)數(shù)由(14.09±2.02)減少至(8.64±1.88),(P0.05)；PKG阻斷劑KT5823同樣抑制了NE的興奮效應(yīng),放電個(gè)數(shù)由(15.56±2.07)減少到(9±0.75),(P0.05)。 結(jié)論： 1、 CCD后部分DRG小細(xì)胞出現(xiàn)交感敏化現(xiàn)象。 2、交感敏化現(xiàn)象中去甲腎上腺素作用受體為α2受體。 3、NO-cGMP-PKG信號(hào)通路參與了交感敏化現(xiàn)象的發(fā)生。大量文獻(xiàn)報(bào)道，受損DRG神經(jīng)元的異位自發(fā)放電與神經(jīng)病理性痛行為密切相關(guān)，關(guān)于異位自發(fā)放電的通道電流機(jī)制也有大量研究，研究結(jié)果顯示多種通道電流大小的改變是神經(jīng)病理痛發(fā)生的一個(gè)重要因素。但迄今為止，還沒(méi)有文獻(xiàn)報(bào)道某種特定的放電模式（如周期放電、burst等）有著與其對(duì)應(yīng)的疼痛特征，如外周神經(jīng)損傷后的痛覺(jué)過(guò)敏、觸誘發(fā)痛和自發(fā)痛等，因此，很難將通道電流的變化與痛信號(hào)模式的關(guān)系作出準(zhǔn)確判斷，影響了痛信號(hào)產(chǎn)生機(jī)制的深入研究。 目的：本論文的第二部分實(shí)驗(yàn)擬在證明“誘發(fā)簇放電（Evoked bursting, EB）”可能是觸誘發(fā)痛的痛信號(hào)的基礎(chǔ)上，對(duì)EB現(xiàn)象及其它神經(jīng)病理性痛的通道電流機(jī)制進(jìn)行深一步的研究。方法：以正常大鼠和慢性背根節(jié)壓迫大鼠為實(shí)驗(yàn)對(duì)象，利用離體整節(jié)DRG標(biāo)本及膜片鉗技術(shù)，對(duì)EB及可能的交感敏化的離子通道機(jī)制進(jìn)行深入探討。 結(jié)果： α-DTX敏感鉀電流（IDTX）參與介導(dǎo)誘發(fā)簇放電（EB）發(fā)生。 α-DTX是低閾值K+通道的阻斷劑，對(duì)Kv1.1、1.2高度敏感。在CCD后的DRG大神經(jīng)元上應(yīng)用1nM的α-DTX使得神經(jīng)元興奮性顯著增加，EB的串長(zhǎng)增加（n=5），部分神經(jīng)元還出現(xiàn)自發(fā)放電活動(dòng)（n＝2）。 結(jié)論： 在CCD模型中，IDTX參與介導(dǎo)DRG大神經(jīng)元誘發(fā)簇放電（EB）的發(fā)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
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R-332
本文編號(hào)：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
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R-332
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 ;Involvement of hyperpolarization-activated,cyclic nucleotide-gated cation channels in dorsal root ganglion in neuropathic pain[J];生理學(xué)報(bào);2008年05期
2 Judith A.Strong;;Recent evidence for activity-dependent initiation of sympathetic sprouting and neuropathic pain[J];生理學(xué)報(bào);2008年05期
，本文編號(hào)：2306694
本文鏈接：http://sikaile.net/xiyixuelunwen/2306694.html
最近更新
教材專著
