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Microencapsulated Schwann cell transplantation inhibits p2x3 receptor expression in dorsal root ganglia and neuropathic pain

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Neural Regeneration Research 2018年第11期13卷 1961-1967页

作者： Ya-Ling Zhang De-Jian Chen Bao-Lin Yang Tao-Tao Liu Jia-Juan Li xiu-Qi Wang Guo-Yong xue Zeng-xu Liu Department of Anatomy Basic Medical School Nanchang University NanchangJiangxi Province China First Affiliated Hospital of Nanchang University Nanchang jiangxi Province China Fourth Clinical Medical College of Nanchang University Nanchang Jiangxi Province China Second Clinical Medical College of Nanchang University Nanchang Jangxi Province China Queen Mary College of Nanchang University Nanchang Jiangxi Province China

Schwann cell transplantation is a promising method to promote neural repair, and can be used for peripheral nerve protection and myelination. Microcapsule technology largely mitigates immune rejection of transplanted cells. We previously showed that microencapsulated olfactory ensheathing cells can reduce neuropathic pain and we hypothesized that microencapsulated Schwann cells can also inhibit neuropathic pain. Rat Schwann cells were cultured by subculture and then microencapsulated and were tested using a rat chronic constriction injury（CCI） neuropathic pain model. CCI rats were treated with Schwann cells or microencapsulated Schwann cells and were compared with sham and CCI groups. Mechanical withdrawal threshold and thermal withdrawal latency were assessed preoperatively and at 1, 3, 5, 7, 9, 11 and 14 days postoperatively. The expression of p2x3 receptors in L4-5 dorsal root ganglia of the different groups was detected by double-label immunofluorescence on day 14 after surgery. Compared with the chronic constriction injury group, mechanical withdrawal threshold and thermal withdrawal latency were higher, but the expression of p2x3 receptors was remarkably decreased in rats treated with Schwann cells and microencapsulated Schwann cells, especially in the rats transplanted with microencapsulated Schwann cells. The above data show that microencapsulated Schwann cell transplantation inhibits p2x3 receptor expression in L4-5 dorsal root ganglia and neuropathic pain.

关键词： nerve regeneration neuropathic pain peripheral nerve injury sciatic nerve microencapsulation Schwann cells p2x3 receptor dorsal root ganglion chronic constriction injury cell transplantation neural repair neural regeneration

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CaMKIIα and caveolin-1 cooperate to drive ATp-induced membrane delivery of the p2x3 receptor

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Journal of Molecular Cell Biology 2014年第2期8卷 140-153页

作者： xu-Qiao Chen Jing-xiang Zhu Yan Wang xu Zhang Lan Bao State Key Laboratory of Cell Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China Institute of Neuroscience and State Key Laboratory of Neuroscience Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

The p2x3 receptor plays a vital role in sensory processing and transmission. The assembly and trafficking of the p2x3 receptor are important for its function in primary sensory neurons. As an important inflammation mediator, ATp is released from different cell types around primary sensory neurons, especially under pathological pain conditions. Here, we showthat α, β-MeATp dramatically promoted membrane delivery of the p2x3 receptor both in HEK293T celts expressing recombinant p2x3 receptor and in rat primary sensory neurons. α, β-MeATp induced p2x3 receptor-mediated Ca^2＋ influx, which further activated Ca^2＋/calmodulin-dependent protein kinase Ilec （CaMKIIα）. The N terminus of the p2x3 receptor was responsible for CaMKIleα binding, whereas Thr38s in the C terminus was phosphorylated by CaMKIIα. Thr^388 phosphorylation increased p2x3 receptor binding to caveoUn-1. CaveoUn-1 knockdown abrogated the α, β-MeATp-induced membrane insertion of the p2x3 receptor. Moreover,α, β-MeATp drove the CaMKIlec-mediated membrane coinsertion of the p2x2 receptor with the p2x3 receptor. The increased p2x3 receptors on the cell membrane that are due to Thr388 phosphorytation facilitated p2x3 receptor-mediated signal transduction. Together, our data indicate that CaMKIIoL and caveoUn-1 cooperate to drive Ugand-induced membrane delivery of the p2x3 receptor and may provide a mechanism of p2x3 receptor sensitization in pain development.

关键词： p2x3 receptor ATp membrane delivery CaMKIIα caveolin-1

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Effect of emodin on neuropathic pain transmission mediated by p2x3 receptor and CGRp of tri-geminal ganglion

Effect of emodin on neuropathic pain transmission mediated b...

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中国生理学会第24届全国会员代表大会暨生理学学术大会

作者：熊伟吴琴黄莉萍高云梁尚栋南昌大学附属口腔医院南昌大学基础医学院

The present research investigated the effect of emodin on trigeminal neuralgia mediated by p2x3 receptor and *** trigeminal neuralgia rat model（a chronic constriction injury of the infraorbital branch of the trigeminal nerve,CCI-ION）was *** male Sprague-Dawley rats were randomly assigned to five groups for varied purpose:blank control group（Control）,sham operation group（Sham）,trigeminal neuralgia model group（TN）,TN rats treated with emodin（TN+E）and TN rats treated with A-317491（TN+A）.The mechanical hyperalgesia threshold of trigeminal neuralgia（TN）rats was tested by the von Frey *** change of the expression of p2x3 receptor and CGRp in rats’trigeminal ganglion（TG）was detected with RT-pCR,immunohistochemical staining,immunofluorescence,Western blotting.14 days afteroperation,the threshold of mechanical hyperalgesia for the territory of ligated infraorbital nerve and the expression of p2x3 receptor and CGRp in TN group were significantly decreased compared with those in Control group,Sham group,TN+E group or TN+A *** responses of mechanical hyperalgesia and the expressions of p2x3 receptor and CGRp in TN+E group were lower than those in TN ***,emodin might inhibit the expression and activation of p2x3 receptor and CGRp in TG to relieve trigeminal neuralgia.

关键词： Trigeminal neuralgia p2x3 receptor CGRp emodin

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Effect of mlncRNA2Y on the expression of p2x3 receptor in primary cultured dorsal root ganglion neurons

Effect of mlncRNA2Y on the expression of P2X3 receptor in pr...

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中国神经科学学会第十届全国学术会议

作者： Chaoran Zheng Shicheng Yu Shuangmei Liu Haiying peng Shangdong Liang Guilin Li Department of physiology Medical College of Nanchang University

Objective p2x receptor was related to the transmission of nociceptive *** expression of mlncRNAs and factors influencing mlncRNAs expression were associated with the onset of nervous system’s *** present study is aimed to explore the effect of mlncRNA2Y on the expression of p2x receptor in primary cultured dorsal root ganglion(DRG) *** Cellular inflammatory pain model was induced by treatment with different concentration of bradykinin for 2 *** the bradykinin and siRNA group,bradykinin was given after transfection of mlncRNA2Y *** expression levels of p2x mRNA and protein were analyzed by real-time pCR and immunofluorescence cytochemistry in DRG *** The levels of mlncRNA2Y and p2x mRNA were increased in primary cultured DRG neurons treated with bradykinin for 2 ***,the expression of p2xreceptor was decreased after knockdown of mlncRNA2Y expression by transfection with *** MlncRNA2Y could up-regulate the expression of p2x receptor in DRG neurons’ inflammatory pain model induced by bradykinin.

关键词： p2x3 receptor dorsal root ganglia inflammatory pain

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The study of p2x3 receptor role in neonatal rat DRG neurons in pain development

The study of P2X3 receptor role in neonatal rat DRG neurons ...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Lu DU,Wen-Juan HE,Jian CUI,Yan-Dong ZHAO,Huai-Zhen RUAN~* Department of Neurobiology,College of Basic Medical Sciences,Chongqing Key Laboratory of Neurobiology,Third Military Medical University,Chongqing 400038,China

＜正＞Objective p2x3 receptor plays important role in the onset and persistence of pain,recent study have shown that p2x3 receptor also plays roles during individual *** we want to figure out whether pain in neonatal rat affects the expression level of p2x3 receptor in primary sensory neurons during development,and explore the potential mechanisms *** Rats of postnatal 3,7,14,21,28 d and adult rats all received CCI （chronic constriction injury） operation in the left sciatic nerve,the right nerve received sham *** threshold （TWL and pWT） was assayed by thermal radiation and von Frey filament before and 1,3,7,14,21,28 days after *** of p2x3 receptor in DRG of every group was measured by immunohistochemical staining（IHC）, and the expression level was *** statistical calculations were performed usingthe SpSS statistical package（SpSS,Chicago,USA）.Results（1） After operation,thermal hyperalgesia and mechanical allodynia appeared on the neonatal and adult rats,and the change pattern of pain threshold in neonatal and adult rats is similar. The TWL and pWT decreased obviously from 1 d,descended to the lowest at 14 d post-operation,then returned to the normal level after 28 d post-operation.（2） The expression of p2x3 receptor in adult rat DRG increased obviously from 1 day post-operation and lasted for 21 days,and returned to the normal level after 28 days;while the expression of p2x3 receptor in neonatal rats DRG dropped obviously in all *** p2x3 receptor plays an important role in the incidence and development of pain and nervous *** the result suggests that pain stimulus might affect the development of neonatal nervous system.

关键词： DRG pain p2x3 receptor development

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Effects of puerarin on burn injury pain mediated by p2x3 receptor

Effects of puerarin on burn injury pain mediated by P2X3 rec...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Chang-Shui xU,Jia-Ri LIN,Shuang-Mei LIU,Shang-Dong LIANG~* Department of physiology,School of Basic Medical Sciences of Nanchang University,Nanchang 330006,China

Objective peripheral administration of p2x agonists rapidly causes nocifensive behavior in experimental animals and pain sensation in *** of the nociceptive response to peripheral ATp is mediated by p2x *** shows that puerarin may have anti-nociceptive response and the role of *** effects of puerarin on rat burn injury pain mediated by p2x receptor was investigated at the present *** Superficial second degree foot/back burn injury models were ***-Dawley rat were randomly divided into sham paw burn group(SpB),puerarin-treated paw unburned control group(ppC),untreated superficial second degree paw burn group(UpB),puerarin-treated superficial second degree paw burn group(ppB),blank paw control group(BpC group),sham back burn group(SBB),puerarin-treated back unburned control group(pBC),untreated superficial second degree back burn group(UBB),puerarin-treated superficial second degree back burn group(pBB) and blank back control group(BBC).Mechanical withdrawal threshold(MWT)and thermal withdrawal latency (TWL)were measured and the expressions of p2x receptor in dorsal root ganglion from test rats were detected by immunohisto-chemistry,RT-pCR,Western blot and in situ *** MWT and TWL in group UpB and ppB were lower than those in group SpB,ppC and BpC(pp ppB were increased compared with those in group UpB after burn 48 h and 1 h respectively(pp ppB and group SpB(p>0.05),and there also was no difference in MWT and TWL between ppC and BpC(p>0.05).The MWT and TWL in group UpB were still decreased,there was difference between group UpB and group SpB(pppC and BpC (ppost burn,the expressions of p2x protein and mRNA in dorsal root ganglion from burn injury model rat in group UpB were increased significantly compared with those in other gro

关键词： p2x3 receptor puerarin burn pain

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Lipid rafts mediates ATp-induced endocytosis,signal transduction and retrograde transport of p2x3 receptor

Lipid rafts mediates ATP-induced endocytosis,signal transduc...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： xu-Qiao CHEN~(1,#),Bin WANG~(1,#),Chengbiao WU~2,Jin pAN~1,Bo YUAN~3,Yuan-Yuan SU~1,xing-Yu JIANG~3,xu ZHANG~4 Lan BAO~(1,*) 1 Laboratory of Molecular Cell Biology,Institute of Biochemistry and Cell Biology,Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences,Shanghai 200031,China 2 Department of Neurosciences,School of Medicine,University of California,San Diego,La Jolla,California,USA 3National Center of Nanoscience and Technology,Beijing 100190,China 4 Institute of Neuroscience and State Key Laboratory of Neuroscience,Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences,Shanghai 200031,China

p2x receptors are a family of cation channels expressed in neurons and nonneuronal *** play a variety of roles including synaptic transmission,muscle contraction,neuronal secretion and inflammation response. Among p2x receptors,the p2x subtype is selectively expressed in dorsal root ganglion *** raft is a specialized membrane microdomain and participate many signal transduction *** report showed that p2x receptor is localized in the lipid rafts of DRG *** have found p2x receptor engaged in retrograde transport in the form of signaling *** we investigated the role of lipid rafts in a,B-MeATp-dependent endocytosis and signal transduction,and retrograde transport of p2x *** of lipid raft with MBCD or nystatin abolished ATp-dependent endocytosis of p2x receptor and ATp-induced ERK activation in p2x receptor-expressing HEK293 *** vivo sucrose gradient fractionation with vesicular pellets prepared from sciatic nerve axoplasm showed that abundant p2x receptor was localized in lipid raft *** transported p2x receptors were colocalized with the lipid raft marker caveolinla in sciatic *** terminal treatment with MpCD attenuated retrograde transport of p2x receptor in axon of dorsal root ganglion ***,lipid raft mediates ATp-induced endocytosis,signal transduction and retrograde transport of p2x receptor.

关键词： p2x3 receptor lipid raft endocytosis ERK retrograde transport

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Electroacupuncture and A-317491 depress the transmission of pain on primary afferent mediated by p2x3 receptor in rats of chronic neuropathic pain states

Electroacupuncture and A-317491 depress the transmission of ...

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2015浙江省物理医学与康复学学术年会

作者：王万胜屠文展蒋松鹤 Rui-Dong Cheng Rong He Li-Hua Ruan Li Zhang Yong-Sheng Gong xiao-Fang Fan Jie Hu Bo Cheng Yin-ping Lai En-Miao Zou Department of physical Medicine and Rehabilitation The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University Department of Rehabilitation Medicine Zhejiang Provincial People's Hospital Institute of Hypoxia Medical Research Teaching Center of Functional ExperimentWenzhou Medical University

p2x is a family of ligand-gated ion channels that act through adenosine ATp.p2x

关键词： Electroacupuncture A-317491 p2x3 receptor Neuropathic pain Dorsal root ganglia Spinal cord

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Effect of mlnc RNA2Y on the neuropathic pain mediated by p2x3 receptor in dorsal root gangli-on

Effect of mlnc RNA2Y on the neuropathic pain mediated by P2X...

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中国生理学会第24届全国会员代表大会暨生理学学术大会

作者： Huaide Jiang Chaoran Zheng Shangdong Liang Guilin Li Department of physiology Medical College of Nanchang University

Objective:Neuropathic pain is an intractable *** pathogenesis is associated with the abnormalities of gene transcription and translation in the sensory *** RNA like non-coding RNAs（mlnc RNAs）as a new member of gene regulation is being more and more attention,but how mlnc RNAs regulate the neuropathic pain is unclear.p2x3 receptor is closely related to the development of neuropathic *** study will be to explore the effect and possible mechanisms of m RNA-like non-coding RNA2Y（mlnc RNA2Y）on p2x3 receptor-mediated neuropathic pain by sciatic chronic constriction injury（CCI）rats ***:The changes of the paw thermal latency（TWL）and the mechanical withdrawal threshold（MWT）were *** expression levels of p2x3 receptor in dorsal root ganglion（DRG）were detected by Western Blot and Real-time quantitative *** expression levels of mlnc RNA2Y in DRG were detected by quantitative *** The behaviorresults of CCI rats showed that at 5 days after surgery,TWL and MWT on surgical side limbs of CCI operated rats were much lower than those in sham rats（p＜0.05）.Compared with the CCI rats,TWL and MWT in CCI treated with mlnc RNA2Y si RNA or A-317491 rats were increased continuously during 7-14days of post-operation（p＜0.01）,in contrast,TWL and MWT in control transfected with mlnc RNA2Y plasmid rats were *** pCR results showed that the expressions of mlnc RNA2Y and p2x3 in DRG of CCI rats and control transfected with mlnc RNA2Y plasmid rats were higher than those in sham rats,but the elevated expressions of mlnc RNA2Y and p2x3 in CCI rats treated with mlnc RNA2Y si RNA were decreased in comparison with those in CCI *** blot results showed that compared with sham rats,the expression levels of p2x3 protein in L4/L5 DRG of CCI and control transfected with mlnc RNA2Y plasmid rats were increased（p＜0.05）.However p2x3 expressions in CCI treated with mlnc RNA2Y si RNA or A-317491 rats were lower than those in CC

关键词： p2x3 receptor dorsal root ganglia neuropathic pain mlncRNA2Y

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Effects of tetramethylpyrazine on trigeminal neuralgia mediated by p2x3 receptor

Effects of tetramethylpyrazine on trigeminal neuralgia media...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Rao-ping WU,Han LIU,Shu-Yi QIU,Yun GAO~*,Shang-Dong LIANG Department of physiology,Basic Medical College of Nanchang University,Nanchang 330006,China

Objective Trigeminal neuralgia(TN)is an uncommon disorder characterized by recurrent attacks of lancinating pain in the trigeminal nerve ***(TMp)is one of the alkaloids contained in Ligustrazine which has been used in traditional Chinese medicine as an analgesic for *** study was designed to explore the effects of TMp on the expression of p2x receptor in trigeminal ganglia(TG)of rats with trigeminal neuralgia(TN).Methods Sprague-Dawley rats were randomly divided into Sham group,TN group and TMp + TN *** right unilateral TN of rat was produced by placing loose chromic gut ligature around the infraorbital nerve (ION).The expression changes of p2x receptor in TG were detected by immunohistochemistry,Western blotting and RT-pCR on day 15 after *** Mechanical response threshold was observed before operation and 3, 5,7,9,11,13,15 days after *** TN rats,an allodynia in response to mechanical stimulation of the ligated nerve territory was found on post-operative day 9-15(pp significantly decreased behavioral signs of neuropathic pain from the 8th day after *** with the sham rats,the expression of the TG p2x in the TN group rats was significantly increased(pxpression of TG p2x in the TMp + TN group rats is lower than that in the TN group rats(pp could reduce the expression of p2x in trigeminal neurons in TN *** may indicate that TMp may inhibit the pain transmission of neuropathic pain through down-regulated p2x receptor in the trigeminal ganglia of TN rats.

关键词： trigeminal neuralgia p2x3 receptor tetramethylpyrazine

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