蛋白酪氨酸磷酸酶SHP-2在Ⅰ型代谢型谷氨酸受体诱发痛觉超敏中的作用
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摘要
目的探讨Ⅰ型代谢型谷氨酸受体(mGluRs)的痛觉调制作用与蛋白酪氨酸磷酸酶SHP-2(Src homology-2 domain-containing phosphatase-2)的关系。方法以免疫共沉淀法,研究小鼠脊髓背角SHP-2与Ⅰ型mGluRs之间的相互作用;通过测定缩足阈值,观察SHP-2抑制剂NSC-87877或SHP-2的无活性突变体SHP-2(C459S)对Ⅰ型mGluRs激动剂DHPG(50 nmol)诱发的痛觉超敏的影响。结果抗m GluR5的特异性抗体,能够从脊髓背角中免疫共沉淀SHP-2,而抗mGluR1的特异性抗体则无法沉淀出SHP-2;以NSC-87877(6 nmol)或SHP-2(C459S)抑制SHP-2的活性,可有效缓解DHPG诱发的痛觉超敏。结论 SHP-2与mGluR5存在分子间的相互结合,SHP-2的激活可能参与Ⅰ型mGluRs的痛觉调制过程。
Aim To investigate whether the pain modification by group Ⅰ metabotropic glutamate receptors(mGluRs) required the involvement of Src homology-2domain-containing phosphatase-2( SHP-2). Methods Co-immunoprecipitation was performed to examine the possible interaction between SHP-2 and group I mGluRs in spinal cord dorsal horn of mice. By measuring the paw withdrawal thresholds,the effects of SHP-2inhibitor NSC-87877 or its catalytically inactive SHP-2( C459S) mutant on allodynia induced by group Ⅰ mGluRs agonist DHPG(50 nmol) were observed. Results Anti-mGluR5 antibody was able to co-immunoprecipitate SHP-2 from spinal dorsal horn of mice,while no SHP-2 was precipitated by anti-mGluR1 antibody. Inactivation of SHP-2 by NSC-87877(6 nmol)or SHP-2( C459S) effectively attenuated allodynia caused by DHPG. Conclusion SHP-2 can physically interact with m GluR5. The activation of SHP-2 may be necessary for group Ⅰ mGluRs to process the nociceptive information.
Aim To investigate whether the pain modification by group Ⅰ metabotropic glutamate receptors(mGluRs) required the involvement of Src homology-2domain-containing phosphatase-2( SHP-2). Methods Co-immunoprecipitation was performed to examine the possible interaction between SHP-2 and group I mGluRs in spinal cord dorsal horn of mice. By measuring the paw withdrawal thresholds,the effects of SHP-2inhibitor NSC-87877 or its catalytically inactive SHP-2( C459S) mutant on allodynia induced by group Ⅰ mGluRs agonist DHPG(50 nmol) were observed. Results Anti-mGluR5 antibody was able to co-immunoprecipitate SHP-2 from spinal dorsal horn of mice,while no SHP-2 was precipitated by anti-mGluR1 antibody. Inactivation of SHP-2 by NSC-87877(6 nmol)or SHP-2( C459S) effectively attenuated allodynia caused by DHPG. Conclusion SHP-2 can physically interact with m GluR5. The activation of SHP-2 may be necessary for group Ⅰ mGluRs to process the nociceptive information.
引文
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[10]Mao L M,Wang J Q.Tyrosine phosphorylation of glutamate receptors by non-receptor tyrosine kinases:roles in depression-like behavior[J].Neurotransmitter(Houst),2016,3:e1118.
[11]Orlando L R,Dunah A W,Standaert D G,Young A B.Tyrosine phosphorylation of the metabotropic glutamate receptor m GluR5 in striatal neurons[J].Neuropharmacology,2002,43(2):161-73.
[12]Mao L M,Liu X Y,Zhang G C,et al.Phosphorylation of group I metabotropic glutamate receptors(m GluR1/5)in vitro and in vivo[J].Neuropharmacology,2008,55(4):403-8.
[2]Suo Z W,Yang X,Li L,et al.Inhibition of protein tyrosine phosphatases in spinal dorsal horn attenuated inflammatory pain by repressing Src signaling[J].Neuropharmacology,2013,70:122-30.
[3]Yang J,Liu L,He D,et al.Crystal structure of human proteintyrosine phosphatase SHP-1[J].J Biol Chem,2003,278(8):6516-20.
[4]Poole A W,Jones M L.A SHPing tale:perspectives on the regulation of SHP-1 and SHP-2 tyrosine phosphatases by the C-terminal tail[J].Cell Signal,2005,17(11):1323-32.
[5]Cho C H,Shin H K.Spinal metabotropic glutamate receptors(m GluRs)are involved in the melittin-induced nociception in rats[J].Korean J Physiol Pharmacol,2008,12(5):237-43.
[6]Conn P J,Pin J P.Pharmacology and functions of metabotropic glutamate receptors[J].Annu Rev Pharmacol Toxicol,1997,37:205-37.
[7]Ango F,Pin J P,Tu J C,et al.Dendritic and axonal targeting of type 5 metabotropic glutamate receptor is regulated by homer1 proteins and neuronal excitation[J].J Neurosci,2000,20(23):8710-6.
[8]Valerio A,Rizzonelli P,Paterlini M,et al.m GluR5 metabotropic glutamate receptor distribution in rat and human spinal cord:a developmental study[J].Neurosci Res,1997,28(1):49-57.
[9]Chen L,Sung S S,Yip M L,et al.Discovery of a novel SHP2protein tyrosine phosphatase inhibitor[J].Mol Pharmacol,2006,70(2):562-70.
[10]Mao L M,Wang J Q.Tyrosine phosphorylation of glutamate receptors by non-receptor tyrosine kinases:roles in depression-like behavior[J].Neurotransmitter(Houst),2016,3:e1118.
[11]Orlando L R,Dunah A W,Standaert D G,Young A B.Tyrosine phosphorylation of the metabotropic glutamate receptor m GluR5 in striatal neurons[J].Neuropharmacology,2002,43(2):161-73.
[12]Mao L M,Liu X Y,Zhang G C,et al.Phosphorylation of group I metabotropic glutamate receptors(m GluR1/5)in vitro and in vivo[J].Neuropharmacology,2008,55(4):403-8.