Cx43及GLT-1对三叉神经痛大鼠疼痛行为学变化的调控作用
|
摘要
目的:研究大鼠三叉神经痛模型中缝隙连接蛋白43(connexin-43,Cx43)和谷氨酸转运体-1(glutamate transporter-1,GLT-1)表达的变化,探讨Cx43和GLT-1与三叉神经痛大鼠疼痛行为的关系。方法:将42只SD大鼠随机分为正常组(N)、假手术组(S)、模型组(ION-CCI)、ION-CCI+生理盐水1组(NS1)、ION-CCI+Gap26组(Gap26)、ION-CCI+生理盐水2组(NS2)、ION-CCI+头孢曲松组(Cef),每组6只。用铬肠线疏松结扎大鼠眶下神经建立三叉神经痛动物模型;造模成功后,相应组于术后第8天分别腹腔注射Gap26、头孢曲松或生理盐水;假手术组仅暴露神经,不结扎。于术前1天,术后1、3、5、7、9、11、14天行机械痛阈值及视频行为学检测。术后第15天取三叉神经脊束核,应用蛋白质印迹检测Cx43和GLT-1蛋白表达水平。结果:腹腔注射Gap26或头孢曲松可提高三叉神经痛大鼠的机械痛阈值(P<0.001),减少其抓脸次数(P<0.001)。术后第15天,ION-CCI、NS1及NS2组大鼠较N组及S组大鼠相比,Cx43表达明显增加(P<0.01),GLT-1表达明显减少(P<0.001);Gap26组大鼠较ION-CCI、NS1组大鼠相比,Cx43表达减少(P<0.05),GLT-1表达增加(P<0.01);Cef组大鼠较ION-CCI、NS2组大鼠相比,GLT-1表达增加(P<0.001),Cx43表达无明显差异。结论:腹腔注射Gap26或头孢曲松可改变三叉神经痛大鼠疼痛行为并改变三叉神经脊束核中Cx43和GLT-1的表达,提示Cx43和GLT-1参与了三叉神经痛大鼠疼痛行为的变化。
Objective:To investigate the expression of connexin-43(Cx43) and glutamate transporter-1(GLT-1),and the relationship among Cx43,GLT-1 and the pain behaviors of trigeminal neuralgia model of rats.Methods:Forty-two Sprague-Dawley rats were randomly divided into Normal(N),Sham(S),Model(ION-CCI),ION-CCI + normal saline 1(NS1),ION-CCI + Gap26(Gap26),ION-CCI + normal saline 2(NS2),and ION-CCI + ceftriaxone groups(Cef),with 6 rats in each group.Rat model of the trigeminal neuralgia was established by ligating the inferior orbital nerve with a chromium enteric line.Inferior orbital nerve was only exposed without ligation in S group.Gap26,ceftriaxone or normal saline was intraperitoneally injected into different groups at the 8 th day after surgery.The mechanical pain threshold and video behavior were detected at 1 day before surgery and 1,3,5,7,9,11,14 days after surgery.The spinal trigeminal nucleus was dissected on postoperative day 15.The expression levels of Cx43 and GLT-1 protein were detected by Western Blot.Results:Intraperitoneal injection of Gap26 or ceftriaxione increased mechanical pain threshold(P < 0.001) and reduced the time of face-grooming(P < 0.001) in ION-CCI rats.On the 15 th day after surgery,compared with N and S group,the expression of Cx43 or GLT-1 were significantly increased(P < 0.01) or reduced(P < 0.001) separately in ION-CCI,NS1 and NS2 groups.Compared with ION-CCI and NS1 group,the expression of Cx43 in Gap26 group was decreased(P < 0.05),and the expression of GLT-1 was increased(P < 0.01).Compared with ION-CCI and NS2 group,the expression of GLT-1 in Cef group was increased(P < 0.001),and there was no significant difference in Cx43 expression.Conclusion:Intraperitoneal injection of Gap26 or ceftriaxone can change the pain behavior in ION-CCI rats and change the expression of Cx43 and GLT-1 in the spinal trigeminal nucleus,suggesting that Cx43 and GLT-1 were involved in the changes of pain behavior of the trigeminal neuralgia in rats.
Objective:To investigate the expression of connexin-43(Cx43) and glutamate transporter-1(GLT-1),and the relationship among Cx43,GLT-1 and the pain behaviors of trigeminal neuralgia model of rats.Methods:Forty-two Sprague-Dawley rats were randomly divided into Normal(N),Sham(S),Model(ION-CCI),ION-CCI + normal saline 1(NS1),ION-CCI + Gap26(Gap26),ION-CCI + normal saline 2(NS2),and ION-CCI + ceftriaxone groups(Cef),with 6 rats in each group.Rat model of the trigeminal neuralgia was established by ligating the inferior orbital nerve with a chromium enteric line.Inferior orbital nerve was only exposed without ligation in S group.Gap26,ceftriaxone or normal saline was intraperitoneally injected into different groups at the 8 th day after surgery.The mechanical pain threshold and video behavior were detected at 1 day before surgery and 1,3,5,7,9,11,14 days after surgery.The spinal trigeminal nucleus was dissected on postoperative day 15.The expression levels of Cx43 and GLT-1 protein were detected by Western Blot.Results:Intraperitoneal injection of Gap26 or ceftriaxione increased mechanical pain threshold(P < 0.001) and reduced the time of face-grooming(P < 0.001) in ION-CCI rats.On the 15 th day after surgery,compared with N and S group,the expression of Cx43 or GLT-1 were significantly increased(P < 0.01) or reduced(P < 0.001) separately in ION-CCI,NS1 and NS2 groups.Compared with ION-CCI and NS1 group,the expression of Cx43 in Gap26 group was decreased(P < 0.05),and the expression of GLT-1 was increased(P < 0.01).Compared with ION-CCI and NS2 group,the expression of GLT-1 in Cef group was increased(P < 0.001),and there was no significant difference in Cx43 expression.Conclusion:Intraperitoneal injection of Gap26 or ceftriaxone can change the pain behavior in ION-CCI rats and change the expression of Cx43 and GLT-1 in the spinal trigeminal nucleus,suggesting that Cx43 and GLT-1 were involved in the changes of pain behavior of the trigeminal neuralgia in rats.
引文
[1]Li Hua Hang,Shu Na Li,Luo Hong,et al.Connexin 43Mediates Cx CL12 Production from Spinal Dorsal Horn to Maintain Bone Cancer Pain in Rats.Neurochem Res,2016,41(5):1200~1208.
[2]Xu Qian,Yong Kwan Cheong,Shao Qiu He,et al.Suppression of spinal connexin 43 expressionattenuates mechanical hypersensitivity in rats after an L5 spinal nerve injury.Neuroscience Letters,2014,566(18):194.
[3]Ru Rong Ji,Temugin Berta,Maiken Nedergaard.Glia and pain:Is chronic pain agliopathy.Pain,2013,154(Supp 1):S10~S28.
[4]Peter T Ohara,Jean-Philippe Vit,Aditi Bhargava,et al.Evidence for a role of connexin 43 in trigeminal pain using RNA interference in vivo.J Neurophy-siol,2008,100(6):3064~3073.
[5]Unger T,Bette S,Zhang J,et al.Connexin-deficiency affects expression levels ofglial glutamate transporters within the cerebrum.Neurosci Lett,2012,506(1):12~16.
[6]Zeng J,Cui LY,Feng Y,et al.Electroacupuncture relieves neuropathic pain via upregulation of glutamate transporters in the spinal cord of rats.Neurosci Lett,2016,620:38~42.
[7]Weng HR,Gao M,Maixner DW.Glycogen synthase kinase 3 beta regulates glial glutamate transporter protein expression in the spinal dorsal horn in rats with neuropathic pain.Exp Neurol,2014,252:18~27.
[8]Ramos KM,Lewis MT,Morgan KN,et al.Spianl upregulation of glutamate transporter GLT-1 by ceftriaxone:therapeutic efficacy in a range of experimental nervous system disorders.Neuroscience,2010,169(4):1888~1900.
[9]罗裕辉,张德仁.神经病理性疼痛脊髓背角GLT-1及GLAST表达的变化.中国疼痛医学杂志,2014,20(3):150~159.
[10]Melanie K,Robert G,Luc J,et al.Chronic Constriction Injury of the Infraorbital Nerve in the Rat using modified syringe needle.J Neurosci Methods,2008,172(1):43~47.
[11]Vos BP,Strassman AM,Maciewicz RJ.Behavioral evidence of trigeminal neuropathic pain following chronic constriction injury to the rat's infraorbital nerve.J Neurosi,1994,14(5):2708~2723.
[12]Kajander KC,Bennett GJ.Onset of a painful peripheral neuropathy in rat:A partial and differential deafferentation and spontaneous discharge in Aβand Aδprimary afferent neurons.Neuropysiol,1992,68:734~744.
[13]HervéJC,Derangeon M.Gap26-junction-mediated cell-to-cell communication.Cell and Tissue Res,2013,352(1):21~31.
[14]Li QS,Jun LG,Manman Zhao,et al.A novel cognitive impairment mechanism that astrocytic p-connexin 43promotes neuronic autophagy via activation of P2X7R and down-regulation of GLT-1 expression in the hippocampus following-traumatic brain injury in rats.Behavi Brain Res,2015,291:315~324.
[15]Ning Shen,Li-Qiu Mo,Fen Hu,et al.A novel role of spinal astrocytic connexin43:mediating morphine antinociceptive tolerance by activation of NMDA receptors and inhibition of glutamate transporter-1 in rats.CNS Neurosci Ther,2014,20(8):728~736.
[16]Rothstein JD,Patel S,Regan MR,et al.β-lactam antibiotics offer neuroprotection by increasing glutamate transporter expression.Nature,2005,433(7021):73~77.
[17]Pottabathini R,Kumar A,Bhatnagar A,et al.Ameliorative potential of pioglitazone and ceftriaxone alone and in combination in rat model of neuropathic pain:Targeting PPARγ and GLT-1 pathways.Pharmacol Rep,2016,68(1):85~94.
[2]Xu Qian,Yong Kwan Cheong,Shao Qiu He,et al.Suppression of spinal connexin 43 expressionattenuates mechanical hypersensitivity in rats after an L5 spinal nerve injury.Neuroscience Letters,2014,566(18):194.
[3]Ru Rong Ji,Temugin Berta,Maiken Nedergaard.Glia and pain:Is chronic pain agliopathy.Pain,2013,154(Supp 1):S10~S28.
[4]Peter T Ohara,Jean-Philippe Vit,Aditi Bhargava,et al.Evidence for a role of connexin 43 in trigeminal pain using RNA interference in vivo.J Neurophy-siol,2008,100(6):3064~3073.
[5]Unger T,Bette S,Zhang J,et al.Connexin-deficiency affects expression levels ofglial glutamate transporters within the cerebrum.Neurosci Lett,2012,506(1):12~16.
[6]Zeng J,Cui LY,Feng Y,et al.Electroacupuncture relieves neuropathic pain via upregulation of glutamate transporters in the spinal cord of rats.Neurosci Lett,2016,620:38~42.
[7]Weng HR,Gao M,Maixner DW.Glycogen synthase kinase 3 beta regulates glial glutamate transporter protein expression in the spinal dorsal horn in rats with neuropathic pain.Exp Neurol,2014,252:18~27.
[8]Ramos KM,Lewis MT,Morgan KN,et al.Spianl upregulation of glutamate transporter GLT-1 by ceftriaxone:therapeutic efficacy in a range of experimental nervous system disorders.Neuroscience,2010,169(4):1888~1900.
[9]罗裕辉,张德仁.神经病理性疼痛脊髓背角GLT-1及GLAST表达的变化.中国疼痛医学杂志,2014,20(3):150~159.
[10]Melanie K,Robert G,Luc J,et al.Chronic Constriction Injury of the Infraorbital Nerve in the Rat using modified syringe needle.J Neurosci Methods,2008,172(1):43~47.
[11]Vos BP,Strassman AM,Maciewicz RJ.Behavioral evidence of trigeminal neuropathic pain following chronic constriction injury to the rat's infraorbital nerve.J Neurosi,1994,14(5):2708~2723.
[12]Kajander KC,Bennett GJ.Onset of a painful peripheral neuropathy in rat:A partial and differential deafferentation and spontaneous discharge in Aβand Aδprimary afferent neurons.Neuropysiol,1992,68:734~744.
[13]HervéJC,Derangeon M.Gap26-junction-mediated cell-to-cell communication.Cell and Tissue Res,2013,352(1):21~31.
[14]Li QS,Jun LG,Manman Zhao,et al.A novel cognitive impairment mechanism that astrocytic p-connexin 43promotes neuronic autophagy via activation of P2X7R and down-regulation of GLT-1 expression in the hippocampus following-traumatic brain injury in rats.Behavi Brain Res,2015,291:315~324.
[15]Ning Shen,Li-Qiu Mo,Fen Hu,et al.A novel role of spinal astrocytic connexin43:mediating morphine antinociceptive tolerance by activation of NMDA receptors and inhibition of glutamate transporter-1 in rats.CNS Neurosci Ther,2014,20(8):728~736.
[16]Rothstein JD,Patel S,Regan MR,et al.β-lactam antibiotics offer neuroprotection by increasing glutamate transporter expression.Nature,2005,433(7021):73~77.
[17]Pottabathini R,Kumar A,Bhatnagar A,et al.Ameliorative potential of pioglitazone and ceftriaxone alone and in combination in rat model of neuropathic pain:Targeting PPARγ and GLT-1 pathways.Pharmacol Rep,2016,68(1):85~94.