title: Promoted Interaction of Nuclear Factor-κB With Demethylated Purinergic P2X3 Receptor Gene Contributes to Neuropathic Pain in Rats With Diabetes.
abstract: Painful diabetic neuropathy is a common complication of diabetes produced by mechanisms that as yet are incompletely defined. The aim of this study was to investigate the roles of nuclear factor-κB (NF-κB) in the regulation of purinergic receptor P2X ligand-gated ion channel 3 (P2X3R) plasticity in dorsal root ganglion (DRG) neurons of rats with painful diabetes. Here, we showed that hindpaw pain hypersensitivity in streptozocin-induced diabetic rats was attenuated by treatment with purinergic receptor antagonist suramin or A-317491. The expression and function of P2X3Rs was markedly enhanced in hindpaw-innervated DRG neurons in diabetic rats. The CpG (cytosine guanine dinucleotide) island in the p2x3r gene promoter region was significantly demethylated, and the expression of DNA methyltransferase 3b was remarkably downregulated in DRGs in diabetic rats. The binding ability of p65 (an active form of NF-κB) with the p2x3r gene promoter region and p65 expression were enhanced significantly in diabetes. The inhibition of p65 signaling using the NF-κB inhibitor pyrrolidine dithiocarbamate or recombinant lentiviral vectors designated as lentiviral vector-p65 small interfering RNA remarkably suppressed P2X3R activities and attenuated diabetic pain hypersensitivity. Insulin treatment significantly attenuated pain hypersensitivity and suppressed the expression of p65 and P2X3Rs. Our findings suggest that the p2x3r gene promoter DNA demethylation and enhanced interaction with p65 contributes to P2X3R sensitization and diabetic pain hypersensitivity.
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases, Institute of Neuroscience, The Second Affiliated Hospital, Soochow University, Suzhou, People's Republic of China.
authors.forename: Hong-Hong
authors.initials: HH
authors.lastname: Zhang
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases, Institute of Neuroscience, The Second Affiliated Hospital, Soochow University, Suzhou, People's Republic of China guangyinxu@suda.edu.cn huji@suda.edu.cn.
authors.forename: Ji
authors.initials: J
authors.lastname: Hu
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases, Institute of Neuroscience, The Second Affiliated Hospital, Soochow University, Suzhou, People's Republic of China.
authors.forename: You-Lang
authors.initials: YL
authors.lastname: Zhou
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases, Institute of Neuroscience, The Second Affiliated Hospital, Soochow University, Suzhou, People's Republic of China.
authors.forename: Xin
authors.initials: X
authors.lastname: Qin
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases, Institute of Neuroscience, The Second Affiliated Hospital, Soochow University, Suzhou, People's Republic of China.
authors.forename: Zhen-Yuan
authors.initials: ZY
authors.lastname: Song
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases, Institute of Neuroscience, The Second Affiliated Hospital, Soochow University, Suzhou, People's Republic of China.
authors.forename: Pan-Pan
authors.initials: PP
authors.lastname: Yang
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases, Institute of Neuroscience, The Second Affiliated Hospital, Soochow University, Suzhou, People's Republic of China.
authors.forename: Shufen
authors.initials: S
authors.lastname: Hu
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases, Institute of Neuroscience, The Second Affiliated Hospital, Soochow University, Suzhou, People's Republic of China.
authors.forename: Xinghong
authors.initials: X
authors.lastname: Jiang
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases, Institute of Neuroscience, The Second Affiliated Hospital, Soochow University, Suzhou, People's Republic of China guangyinxu@suda.edu.cn huji@suda.edu.cn.
authors.forename: Guang-Yin
authors.initials: GY
authors.lastname: Xu
mesh_terms: D000818:Animals; D018899:CpG Islands / Q000187:drug effects; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation / Q000187:drug effects; D003921:Diabetes Mellitus, Experimental / Q000150:complications* / Q000188:drug therapy; D003929:Diabetic Neuropathies / Q000201:enzymology / Q000378:metabolism* / Q000517:prevention & control; D044127:Epigenesis, Genetic / Q000187:drug effects; D005260:Female; D005727:Ganglia, Spinal / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D006614:Hindlimb; D007004:Hypoglycemic Agents / Q000627:therapeutic use; D007328:Insulin / Q000627:therapeutic use; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D009437:Neuralgia / Q000150:complications / Q000378:metabolism* / Q000517:prevention & control; D009474:Neurons / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000187:drug effects; D058920:Purinergic P2X Receptor Antagonists / Q000627:therapeutic use; D034622:RNA Interference; D017207:Rats, Sprague-Dawley; D058477:Receptors, Purinergic P2X3 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051996:Transcription Factor RelA / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D000097584:DNA Methyltransferase 3B
chemical_list: D004248:DNA (Cytosine-5-)-Methyltransferases; D007004:Hypoglycemic Agents; D007328:Insulin; D009419:Nerve Tissue Proteins; D058920:Purinergic P2X Receptor Antagonists; D058477:Receptors, Purinergic P2X3; D051996:Transcription Factor RelA; D000097584:DNA Methyltransferase 3B; C546410:P2rx3 protein, rat; C498145:Rela protein, rat