AIM: To investigate proteomic changes in spinal cord and dorsal root ganglia (DRG) of rats with trinitrobenzene sulfonic acid (TNBS)-induced colitis. METHODS: The colonic myeloperoxidase (MPO) activity and tumor necro...AIM: To investigate proteomic changes in spinal cord and dorsal root ganglia (DRG) of rats with trinitrobenzene sulfonic acid (TNBS)-induced colitis. METHODS: The colonic myeloperoxidase (MPO) activity and tumor necrosis factor-(TNF- ) level were determined. A two-dimensional electrophoresis (2-DE)-based proteomic technique was used to profile the global protein expression changes in the DRG and spinal cord of the rats with acute colitis induced by intracolonic injection of TNBS. RESULTS: TNBS group showed significantly elevated colonic MPO activity and increased TNF-level. The proteins derived from lumbosacral enlargement of the spinal cord and DRG were resolved by 2-DE; and 26 and 19 proteins that displayed significantly different expression levels in the DRG and spinal cord were identified respectively. Altered proteins were found to be involved in a number of biological functions, such as inflammation/immunity, cell signaling, redox regulation, sulfate transport and cellular metabolism. The over-expression of the protein similar to potassium channel tetramerisation domain containing protein 12 (Kctd 12) and low expression of proteasome subunit type-1 (psma) were validated by Western blotting analysis. CONCLUSION: TNBS-induced colitis has a profound impact on protein profiling in the nervous system. This result helps understand the neurological pathogenesis of inflammatory bowel disease.展开更多
Very little is known about the role of melatonin in the trigeminal system, including the function of melatonin receptor 1. In the present study, adult rats were injected with formaldehyde into the right vibrissae pad ...Very little is known about the role of melatonin in the trigeminal system, including the function of melatonin receptor 1. In the present study, adult rats were injected with formaldehyde into the right vibrissae pad to establish a model of orofacial inflammatory pain. The distribution of melatonin re- ceptor 1 and nicotinamide adenine dinucleotide phosphate diaphorase in the caudal spinal trigeminal nucleus and trigeminal ganglion was determined with immunohistochemistry and histo- chemistry. The results show that there are significant differences in melatonin receptor 1 expression and nicotinamide adenine dinucleotide phosphate diaphorase expression in the trigeminal ganglia and caudal spinal nucleus during the early stage of orofacial inflammatory pain. Our findings sug- gest that when melatonin receptor 1 expression in the caudal spinal nucleus is significantly reduced, melatonin＇s regulatory effect on pain is attenuated.展开更多
Nociceptive signals conveyed to the dorsal horn of the spinal cord by primary nociceptors are subject to extensive modulation by local neurons and by supraspinal descending pathways to the spinal cord before being rel...Nociceptive signals conveyed to the dorsal horn of the spinal cord by primary nociceptors are subject to extensive modulation by local neurons and by supraspinal descending pathways to the spinal cord before being relayed to higher brain centers. Descending modulatory pathways to the spinal cord comprise,among others, noradrenergic, serotonergic, γ-aminobutyric acid(GABA)ergic, and dopaminergic fibers.The contributions of noradrenaline, serotonin, and GABA to pain modulation have been extensively investigated. In contrast, the contributions of dopamine to pain modulation remain poorly understood.The focus of this review is to summarize the current knowledge of the contributions of dopamine to pain modulation. Hypothalamic A11 dopaminergic neurons project to all levels of the spinal cord and provide the main source of spinal dopamine. Dopamine receptors are expressed in primary nociceptors as well as in spinal neurons located in different laminae in the dorsal horn of the spinal cord, suggesting that dopamine can modulate pain signals by acting at both presynaptic and postsynaptic targets. Here, I will review the literature on the effects of dopamine and dopamine receptor agonists/antagonists on the excitability of primary nociceptors, the effects of dopamine on the synaptic transmission between primary nociceptors and dorsal horn neurons, and the effects of dopamine on pain in rodents. Published data support both anti-nociceptive effects of dopamine mediated by D2-like receptors and pro-nociceptive effects mediated by D1-like receptors.展开更多
Objective To investigate whether chronic constriction injury(CCI)of the sciatic nerve of rats could produce alterations in the phosphorylation of cyclic AMP response element binding(CREB)protein in dorsal root ganglia...Objective To investigate whether chronic constriction injury(CCI)of the sciatic nerve of rats could produce alterations in the phosphorylation of cyclic AMP response element binding(CREB)protein in dorsal root ganglia(DRG)and superficial dorsal horn neurons of the spinal cord.Methods Chronic constriction injury(CCI)of the sciatic nerve was employed as a model of neuropathic pain.Thirty-two Sprague-Dawley rats were randomly divided into Na⒍ve,Sham,CCI2w(received CCI for2weeks)and CCI4w(received CCI for4weeks)groups.Hind pawwithdrawal threshold to mechanical stimuli and withdrawal latency to thermal stimuli were used to determine the mechanical and thermal hyperalgesia.Then all the rats were deeply anesthetized and perfused intracardially with paraformaldehyde.The fixed L 4-5 spinal cord and the L 5 DRG ipsilateral to CCI were harvested for fixation.The pCREB-immunoreactive(pCREB-IR)cells in both DRG and superficial dorsal horn neurons were quantified for analysis using immunohistochemistry methods.Results On the14th day after sciatic nerve injury,all the rats exhibited significant mechanical and thermal hyperalgesia.The mechanical withdrawal thresholds to von Frey filament from CCI2w group decreased significantly compared to both baseline values and those of Sham group(P<0.01);Thermal withdwal latencies from CCI2w group decreased significantly compared to both baseline values and those of Sham group(P<0.01).Some rats from Sham group also showed mechanical hyperalgesia compared to both baseline values and those of Na⒍ve group(P<0.01).28days after CCI,both mechanical and thermal hypersensitivity were significantly alleviated,with no statistical significance compared to those of Sham group.On the14th day after CCI,the number of pCREB-IR cells significantly increased in ipsilateral L 5 DRGs and superficial dorsal horns(P<0.01)compared to Sham group.The number of phosphorylated CREB-IR cells in the ipsilateral DRGs from Sham group also increased compared to that of Naive rats(P<0.05).There were no significant statistical differences of numbers of CREB-IR neuron between Sham group and CCI4wgroup.Conclusion CCI increases CREB phosphorylation both in DRG and superficial dorsal horn neurons of the lumbar spinal cord,and may be one of the key molecular mechanisms of central and peripheral sensitization following peripheral nerve injury.展开更多
基金Supported by The Research Grants Council of Hong Kong,RGC-HKBU2/07CThe Hong Kong Jockey Club Institute of Chinese Medicine, JCICM4-07
文摘AIM: To investigate proteomic changes in spinal cord and dorsal root ganglia (DRG) of rats with trinitrobenzene sulfonic acid (TNBS)-induced colitis. METHODS: The colonic myeloperoxidase (MPO) activity and tumor necrosis factor-(TNF- ) level were determined. A two-dimensional electrophoresis (2-DE)-based proteomic technique was used to profile the global protein expression changes in the DRG and spinal cord of the rats with acute colitis induced by intracolonic injection of TNBS. RESULTS: TNBS group showed significantly elevated colonic MPO activity and increased TNF-level. The proteins derived from lumbosacral enlargement of the spinal cord and DRG were resolved by 2-DE; and 26 and 19 proteins that displayed significantly different expression levels in the DRG and spinal cord were identified respectively. Altered proteins were found to be involved in a number of biological functions, such as inflammation/immunity, cell signaling, redox regulation, sulfate transport and cellular metabolism. The over-expression of the protein similar to potassium channel tetramerisation domain containing protein 12 (Kctd 12) and low expression of proteasome subunit type-1 (psma) were validated by Western blotting analysis. CONCLUSION: TNBS-induced colitis has a profound impact on protein profiling in the nervous system. This result helps understand the neurological pathogenesis of inflammatory bowel disease.
基金supported by the National Natural Science Foundation of China,No.81271166,81371107the Natural Science Foundation of Guangdong Province in China,No.10451008901006145
文摘Very little is known about the role of melatonin in the trigeminal system, including the function of melatonin receptor 1. In the present study, adult rats were injected with formaldehyde into the right vibrissae pad to establish a model of orofacial inflammatory pain. The distribution of melatonin re- ceptor 1 and nicotinamide adenine dinucleotide phosphate diaphorase in the caudal spinal trigeminal nucleus and trigeminal ganglion was determined with immunohistochemistry and histo- chemistry. The results show that there are significant differences in melatonin receptor 1 expression and nicotinamide adenine dinucleotide phosphate diaphorase expression in the trigeminal ganglia and caudal spinal nucleus during the early stage of orofacial inflammatory pain. Our findings sug- gest that when melatonin receptor 1 expression in the caudal spinal nucleus is significantly reduced, melatonin＇s regulatory effect on pain is attenuated.
基金supported by internal funds to MP from the Department of Anesthesiology,Stony Brook Medicine,USA
文摘Nociceptive signals conveyed to the dorsal horn of the spinal cord by primary nociceptors are subject to extensive modulation by local neurons and by supraspinal descending pathways to the spinal cord before being relayed to higher brain centers. Descending modulatory pathways to the spinal cord comprise,among others, noradrenergic, serotonergic, γ-aminobutyric acid(GABA)ergic, and dopaminergic fibers.The contributions of noradrenaline, serotonin, and GABA to pain modulation have been extensively investigated. In contrast, the contributions of dopamine to pain modulation remain poorly understood.The focus of this review is to summarize the current knowledge of the contributions of dopamine to pain modulation. Hypothalamic A11 dopaminergic neurons project to all levels of the spinal cord and provide the main source of spinal dopamine. Dopamine receptors are expressed in primary nociceptors as well as in spinal neurons located in different laminae in the dorsal horn of the spinal cord, suggesting that dopamine can modulate pain signals by acting at both presynaptic and postsynaptic targets. Here, I will review the literature on the effects of dopamine and dopamine receptor agonists/antagonists on the excitability of primary nociceptors, the effects of dopamine on the synaptic transmission between primary nociceptors and dorsal horn neurons, and the effects of dopamine on pain in rodents. Published data support both anti-nociceptive effects of dopamine mediated by D2-like receptors and pro-nociceptive effects mediated by D1-like receptors.
文摘Objective To investigate whether chronic constriction injury(CCI)of the sciatic nerve of rats could produce alterations in the phosphorylation of cyclic AMP response element binding(CREB)protein in dorsal root ganglia(DRG)and superficial dorsal horn neurons of the spinal cord.Methods Chronic constriction injury(CCI)of the sciatic nerve was employed as a model of neuropathic pain.Thirty-two Sprague-Dawley rats were randomly divided into Na⒍ve,Sham,CCI2w(received CCI for2weeks)and CCI4w(received CCI for4weeks)groups.Hind pawwithdrawal threshold to mechanical stimuli and withdrawal latency to thermal stimuli were used to determine the mechanical and thermal hyperalgesia.Then all the rats were deeply anesthetized and perfused intracardially with paraformaldehyde.The fixed L 4-5 spinal cord and the L 5 DRG ipsilateral to CCI were harvested for fixation.The pCREB-immunoreactive(pCREB-IR)cells in both DRG and superficial dorsal horn neurons were quantified for analysis using immunohistochemistry methods.Results On the14th day after sciatic nerve injury,all the rats exhibited significant mechanical and thermal hyperalgesia.The mechanical withdrawal thresholds to von Frey filament from CCI2w group decreased significantly compared to both baseline values and those of Sham group(P<0.01);Thermal withdwal latencies from CCI2w group decreased significantly compared to both baseline values and those of Sham group(P<0.01).Some rats from Sham group also showed mechanical hyperalgesia compared to both baseline values and those of Na⒍ve group(P<0.01).28days after CCI,both mechanical and thermal hypersensitivity were significantly alleviated,with no statistical significance compared to those of Sham group.On the14th day after CCI,the number of pCREB-IR cells significantly increased in ipsilateral L 5 DRGs and superficial dorsal horns(P<0.01)compared to Sham group.The number of phosphorylated CREB-IR cells in the ipsilateral DRGs from Sham group also increased compared to that of Naive rats(P<0.05).There were no significant statistical differences of numbers of CREB-IR neuron between Sham group and CCI4wgroup.Conclusion CCI increases CREB phosphorylation both in DRG and superficial dorsal horn neurons of the lumbar spinal cord,and may be one of the key molecular mechanisms of central and peripheral sensitization following peripheral nerve injury.
文摘为探讨神经生长因子(NGF)在哮喘发病中的作用及其神经免疫调节机制,本研究应用免疫组织化学方法检测各组豚鼠下呼吸道和内脏感觉传入部位白介素-1β(IL-1β)免疫反应的变化.用Western blot方法分别检测各组豚鼠肺、C7～T5段脊神经节和相应节段脊髓背角NGF和IL-1β的表达.用Luzex-F实时图像分析系统和凝胶成像分析系统分别对结果进行图像分析.免疫组织化学结果显示(染色反应强度用灰度值表示):生理盐水组与单纯致敏组豚鼠IL-1β阳性免疫反应产物的平均灰度值没有显著差异(P＞0.05);与这两个对照组相比,哮喘豚鼠气道上皮、肺内炎性细胞、血管平滑肌内的神经末梢、C7～T5段脊神经节及相应节段脊髓背角内IL-1β阳性反应产物的平均灰度值均明显降低(P＜0.01);在哮喘+NGF抗体组(鼻腔吸入NGF抗体),上述部位内IL-1β阳性反应产物的平均灰度值均明显高于哮喘组(P＜0.01).Western blot结果显示:与生理盐水组和单纯致敏组比较,哮喘组豚鼠肺、C7～T5段脊神经节及相应节段脊髓样品中IL-1β或NGF目标带的IDV(integrated density value)与内参照IDV的比值均明显升高(P＜0.01),而哮喘+NGF抗体组上述部位样品中IL-1β或NGF目标带的IDV与内参照IDV的比值均明显低于哮喘组(P＜0.01),表明NGF可上调IL-1β的表达.这些结果提示NGF诱发IL-1β的表达可能是NGF参与哮喘发病的机制之一,NGF抗体抑制哮喘时下呼吸道和内脏感觉传入部位IL-1β的表达可能是治疗哮喘的新途径.
