Tongfei Cheng, Shanlong Du, Yi Cao, Ziyan Lu, Yingjun Xu
{"title":"新生儿缺氧缺血的神经保护：褪黑素通过PINK1-Parkin通路靶向NCX1抑制线粒体自噬","authors":"Tongfei Cheng, Shanlong Du, Yi Cao, Ziyan Lu, Yingjun Xu","doi":"10.1007/s10735-025-10601-5","DOIUrl":null,"url":null,"abstract":"<div><h3>Objective</h3><p>Hypoxic ischaemic (HI) damage is a major cause of white matter damage (WMD) in the brains of newborns, especially preterm infants; early neuroprotection is essential to improve cognitive outcomes. This study aimed to investigate the effect of melatonin on nerve injury by inhibiting mitochondrial autophagy.</p><h3>Methods</h3><p>We established a neonatal WMD model through HI induction in postnatal day 3 (P3) Sprague–Dawley (SD) rats. Following four days of intraperitoneal melatonin administration (10 mg/kg/d), temporal changes in expression of sodium-calcium exchanger 1 (NCX1), myelin integrity markers (myelin-associated glycoprotein [MAG]/proteolipid protein [PLP]), and mitophagy-related proteins (microtubule-associated protein 1 light chain 3β [LC3β], PTEN-induced kinase 1 [PINK1], and Parkin RBR E3 ubiquitin-protein ligase [Parkin]) were systematically quantified. Neuronal hyperexcitability was evaluated by whole-cell patch-clamp recordings, whereas myelin pathology was assessed by luxol fast blue (LFB) staining, and mitochondrial ultrastructures were evaluated by transmission electron microscopy. Cognitive recovery was determined using Morris water maze testing at postnatal day 28.</p><h3>Results</h3><p>Our results demonstrated that rats subjected to HI presented biphasic alterations in NCX1 expression, characterised by transient upregulation on day 7 followed by a progressive decline (<i>P</i> < 0.001). Concurrently, expression of mitochondrial autophagy markers (LC3β, PINK1, and Parkin) was significantly increased (<i>P</i> < 0.001). Histological analysis revealed distinct mitochondrial structural damage and autophagosome formation. Electrophysiological measurements revealed increased neuronal excitability (<i>P</i> < 0.05), which was correlated with spatial learning and memory deficits. Although melatonin treatment effectively attenuated these pathological alterations, subsequent pharmacological inhibition of NCX1 via SN6 administration in melatonin-treated rats resulted in the recurrence of mitochondrial ultrastructural abnormalities and the reactivation of autophagic pathways.</p><h3>Conclusion</h3><p>Melatonin attenuated activation of the PINK1-Parkin-dependent mitochondrial autophagy pathway in neonatal rats with HI-induced WMD through mediating the dynamic expression of NCX1. This intervention effectively reduced neuronal hyperexcitability, ameliorated demyelinating lesions, and improved long-term learning and cognitive functions.</p><h3>Clinical trial registration</h3><p>Not applicable.</p></div>","PeriodicalId":650,"journal":{"name":"Journal of Molecular Histology","volume":"56 5","pages":""},"PeriodicalIF":2.2000,"publicationDate":"2025-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s10735-025-10601-5.pdf","citationCount":"0","resultStr":"{\"title\":\"Neuroprotection in neonatal Hypoxia-ischaemia: melatonin targets NCX1 to inhibit mitochondrial autophagy via the PINK1-Parkin pathway\",\"authors\":\"Tongfei Cheng, Shanlong Du, Yi Cao, Ziyan Lu, Yingjun Xu\",\"doi\":\"10.1007/s10735-025-10601-5\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Objective</h3><p>Hypoxic ischaemic (HI) damage is a major cause of white matter damage (WMD) in the brains of newborns, especially preterm infants; early neuroprotection is essential to improve cognitive outcomes. This study aimed to investigate the effect of melatonin on nerve injury by inhibiting mitochondrial autophagy.</p><h3>Methods</h3><p>We established a neonatal WMD model through HI induction in postnatal day 3 (P3) Sprague–Dawley (SD) rats. Following four days of intraperitoneal melatonin administration (10 mg/kg/d), temporal changes in expression of sodium-calcium exchanger 1 (NCX1), myelin integrity markers (myelin-associated glycoprotein [MAG]/proteolipid protein [PLP]), and mitophagy-related proteins (microtubule-associated protein 1 light chain 3β [LC3β], PTEN-induced kinase 1 [PINK1], and Parkin RBR E3 ubiquitin-protein ligase [Parkin]) were systematically quantified. Neuronal hyperexcitability was evaluated by whole-cell patch-clamp recordings, whereas myelin pathology was assessed by luxol fast blue (LFB) staining, and mitochondrial ultrastructures were evaluated by transmission electron microscopy. Cognitive recovery was determined using Morris water maze testing at postnatal day 28.</p><h3>Results</h3><p>Our results demonstrated that rats subjected to HI presented biphasic alterations in NCX1 expression, characterised by transient upregulation on day 7 followed by a progressive decline (<i>P</i> < 0.001). Concurrently, expression of mitochondrial autophagy markers (LC3β, PINK1, and Parkin) was significantly increased (<i>P</i> < 0.001). Histological analysis revealed distinct mitochondrial structural damage and autophagosome formation. Electrophysiological measurements revealed increased neuronal excitability (<i>P</i> < 0.05), which was correlated with spatial learning and memory deficits. Although melatonin treatment effectively attenuated these pathological alterations, subsequent pharmacological inhibition of NCX1 via SN6 administration in melatonin-treated rats resulted in the recurrence of mitochondrial ultrastructural abnormalities and the reactivation of autophagic pathways.</p><h3>Conclusion</h3><p>Melatonin attenuated activation of the PINK1-Parkin-dependent mitochondrial autophagy pathway in neonatal rats with HI-induced WMD through mediating the dynamic expression of NCX1. 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Neuroprotection in neonatal Hypoxia-ischaemia: melatonin targets NCX1 to inhibit mitochondrial autophagy via the PINK1-Parkin pathway
Objective
Hypoxic ischaemic (HI) damage is a major cause of white matter damage (WMD) in the brains of newborns, especially preterm infants; early neuroprotection is essential to improve cognitive outcomes. This study aimed to investigate the effect of melatonin on nerve injury by inhibiting mitochondrial autophagy.
Methods
We established a neonatal WMD model through HI induction in postnatal day 3 (P3) Sprague–Dawley (SD) rats. Following four days of intraperitoneal melatonin administration (10 mg/kg/d), temporal changes in expression of sodium-calcium exchanger 1 (NCX1), myelin integrity markers (myelin-associated glycoprotein [MAG]/proteolipid protein [PLP]), and mitophagy-related proteins (microtubule-associated protein 1 light chain 3β [LC3β], PTEN-induced kinase 1 [PINK1], and Parkin RBR E3 ubiquitin-protein ligase [Parkin]) were systematically quantified. Neuronal hyperexcitability was evaluated by whole-cell patch-clamp recordings, whereas myelin pathology was assessed by luxol fast blue (LFB) staining, and mitochondrial ultrastructures were evaluated by transmission electron microscopy. Cognitive recovery was determined using Morris water maze testing at postnatal day 28.
Results
Our results demonstrated that rats subjected to HI presented biphasic alterations in NCX1 expression, characterised by transient upregulation on day 7 followed by a progressive decline (P < 0.001). Concurrently, expression of mitochondrial autophagy markers (LC3β, PINK1, and Parkin) was significantly increased (P < 0.001). Histological analysis revealed distinct mitochondrial structural damage and autophagosome formation. Electrophysiological measurements revealed increased neuronal excitability (P < 0.05), which was correlated with spatial learning and memory deficits. Although melatonin treatment effectively attenuated these pathological alterations, subsequent pharmacological inhibition of NCX1 via SN6 administration in melatonin-treated rats resulted in the recurrence of mitochondrial ultrastructural abnormalities and the reactivation of autophagic pathways.
Conclusion
Melatonin attenuated activation of the PINK1-Parkin-dependent mitochondrial autophagy pathway in neonatal rats with HI-induced WMD through mediating the dynamic expression of NCX1. This intervention effectively reduced neuronal hyperexcitability, ameliorated demyelinating lesions, and improved long-term learning and cognitive functions.
期刊介绍：
The Journal of Molecular Histology publishes results of original research on the localization and expression of molecules in animal cells, tissues and organs. Coverage includes studies describing novel cellular or ultrastructural distributions of molecules which provide insight into biochemical or physiological function, development, histologic structure and disease processes.
Major research themes of particular interest include:
- Cell-Cell and Cell-Matrix Interactions;
- Connective Tissues;
- Development and Disease;
- Neuroscience.
Please note that the Journal of Molecular Histology does not consider manuscripts dealing with the application of immunological or other probes on non-standard laboratory animal models unless the results are clearly of significant and general biological importance.
The Journal of Molecular Histology publishes full-length original research papers, review articles, short communications and letters to the editors. All manuscripts are typically reviewed by two independent referees. The Journal of Molecular Histology is a continuation of The Histochemical Journal.
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