Netrin-1 (NTN-1) plays a vital role in the progress of nervous system development and inflammatory diseases. However, the role and underlying mechanism of NTN-1 in inflammatory pain (IP) are unclear. BV2 microglia were treated with LPS to mimic the cell status under IP. Adeno-associated virus carrying the NTN-1 gene (AAV-NTN-1) was used to overexpress NTN-1. Complete Freund's Adjuvant (CFA)-induced mouse was recruited as an in vivo model. MTT and commercial kits were utilized to evaluate cell viability and cell death of BV2 cells. The mRNA expressions and secretions of cytokines were measured using the ELISA method. Also, the pyroptosis and activation of BV2 cells were investigated based on western blotting. To verify the role of Rac1/NF-kappaB signaling, isochamaejasmin (ISO) and AAV-Rac1 were presented. The results showed that NTN-1 expression was decreased in LPS-treated BV2 microglia and spinal cord tissues of CFA-injected mice. Overexpressing NTN-1 dramatically reversed cell viability and decreased cell death rate of BV2 microglia under lipopolysaccharide (LPS) stimulation, while the level of pyroptosis was inhibited. Besides, AAV-NTN-1 rescued the activation of microglia and inflammatory injury induced by LPS, decreasing IBA-1 expression, as well as iNOS, IL-1beta and IL-6 secretions. Meanwhile AAV-NTN-1 promoted the anti-inflammation response, including increases in Arg-1, IL-4 and IL-10 levels. In addition, the LPS-induced activation of Rac1/NF-kappaB signaling was depressed by NTN-1 overexpression. The same results were verified in a CFA-induced mouse model. In conclusion, NTN-1 alleviated IP by suppressing pyroptosis and promoting M2 type activation of microglia via inhibiting Rac1/NF-?B signaling, suggesting the protective role of NTN-1 in IP. Keywords: Netrin-1, Inflammatory pain, Pyroptosis, Microglia M2 activation, Rac1/NF-kappaB.

• MeSH
• bolest metabolismus   MeSH
• buněčné linie MeSH
• lipopolysacharidy MeSH
• mikroglie * metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• netrin-1 * metabolismus   MeSH
• neuropeptidy * MeSH
• NF-kappa B * metabolismus   MeSH
• pyroptóza * fyziologie   účinky léků   MeSH
• rac1 protein vázající GTP * metabolismus   MeSH
• signální transdukce * MeSH
• zánět * metabolismus   patologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• lipopolysacharidy MeSH
• netrin-1 * MeSH
• neuropeptidy * MeSH
• NF-kappa B * MeSH
• Ntn1 protein, mouse MeSH Prohlížeč
• rac1 protein vázající GTP * MeSH
• Rac1 protein, mouse MeSH Prohlížeč

Previous findings showed that in mice, complete knockout of activity-dependent neuroprotective protein (ADNP) abolishes brain formation, while haploinsufficiency (Adnp+/-) causes cognitive impairments. We hypothesized that mutations in ADNP lead to a developmental/autistic syndrome in children. Indeed, recent phenotypic characterization of children harboring ADNP mutations (ADNP syndrome children) revealed global developmental delays and intellectual disabilities, including speech and motor dysfunctions. Mechanistically, ADNP includes a SIP motif embedded in the ADNP-derived snippet drug candidate NAP (NAPVSIPQ, also known as CP201), which binds to microtubule end-binding protein 3, essential for dendritic spine formation. Here, we established a unique neuronal membrane-tagged, GFP-expressing Adnp+/- mouse line allowing in vivo synaptic pathology quantification. We discovered that Adnp deficiency reduced dendritic spine density and altered synaptic gene expression, both of which were partly ameliorated by NAP treatment. Adnp+/-mice further exhibited global developmental delays, vocalization impediments, gait and motor dysfunctions, and social and object memory impairments, all of which were partially reversed by daily NAP administration (systemic/nasal). In conclusion, we have connected ADNP-related synaptic pathology to developmental and behavioral outcomes, establishing NAP in vivo target engagement and identifying potential biomarkers. Together, these studies pave a path toward the clinical development of NAP (CP201) for the treatment of ADNP syndrome.

• Klíčová slova
• Drug therapy, Therapeutics,
• MeSH
• aminokyselinové motivy MeSH
• autistická porucha genetika   metabolismus   patologie   patofyziologie   MeSH
• biologické markery metabolismus   MeSH
• buněčná membrána genetika   metabolismus   patologie   MeSH
• chování zvířat MeSH
• dendritické trny metabolismus   patologie   MeSH
• homeodoménové proteiny MeSH
• lidé MeSH
• mikrotubuly genetika   metabolismus   patologie   MeSH
• modely neurologické * MeSH
• mutace MeSH
• myši knockoutované MeSH
• myši MeSH
• naftochinony farmakologie   MeSH
• proteiny nervové tkáně nedostatek   MeSH
• regulace genové exprese MeSH
• synapse metabolismus   patologie   MeSH
• syndrom MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• audiovizuální média MeSH
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Názvy látek
• 2-(3,5-di-tert-butyl-4-hydroxyl)-3-chloro-1,4-naphthoquinone MeSH Prohlížeč
• Adnp protein, mouse MeSH Prohlížeč
• biologické markery MeSH
• homeodoménové proteiny MeSH
• naftochinony MeSH
• proteiny nervové tkáně MeSH