Mechanisms Underlying Abnormal Expression of lncRNA H19 in Neonatal Hypoxic–Ischemic Encephalopathy

Abstract

Objective Hypoxic–ischemic (HI)-related brain injury, especially HI encephalopathy (HIE) is a leading cause of morbidity and disability in newborns. Long noncoding RNAs (lncRNAs) are implicated in the progress of HI brain damage. However, the mechanisms underlying the regulatory effects of lncRNA H19 on autophagy in HIE remain unknown. This study was designed to identify the potential mechanisms involving lncRNA H19 in HIE.

Study Design We selected three HIE newborns and three healthy newborns for neonatal behavioral neurological assessment and screened the differentially expressed lncRNAs by microarray analysis and detected H19 expression in serum. After that, neonatal HIE rats were established and injected with H19 overexpression lentivirus vector or autophagy activator Rapa. The structure and apoptotic levels of brain tissue were observed, and righting reflex and geotaxis reflex were utilized to evaluate the short-term neurological function of HIE rats. The Morris water maze was performed to measure the long-term neurological functions of HIE rats. The binding relationships among H19/miR-19b/protein kinase B3 (Akt3) were verified. Levels of Akt3- and autophagy-related proteins were measured.

Results H19 was upregulated in HIE newborns and rat models. The areas of cerebral infarction and apoptosis in neonatal HIE rats were increased, and the nerve functions were compromised. The overexpression of H19 alleviated nerve damage of neonatal HIE rats, and reduced autophagy of brain tissue. H19 upregulated Akt3 as a miR-29b sponge. The protective effects of overexpression of H19 on brain tissue and nerve functions of neonatal HIE rats were partially reversed by autophagy activator.

Conclusion H19 improved the brain tissue and alleviated nerve damage of neonatal HIE rats by upregulating the Akt3/mTOR pathway as a miR-29b sponge.

Key Points

• H19 overexpression reduces the nerve damage in neonatal HIE rats.

• H19 reduces autophagy in neonatal HIE rats by the miR-29b/Akt3/mTOR axis.

• Autophagy activator reverses the protection of H19 in neonatal HIE.

Keywords

hypoxic–ischemic encephalopathy - LncRNA H19 - autophagy - microRNA-29b - akt3/mTOR pathway

Publikationsverlauf

Eingereicht: 26. März 2020

Angenommen: 21. September 2020

Artikel online veröffentlicht:
27. Oktober 2020

• Reference

• 1 Kurinczuk JJ, White-Koning M, Badawi N. Epidemiology of neonatal encephalopathy and hypoxic-ischaemic encephalopathy. Early Hum Dev 2010; 86 (06) 329-338
• 2 Murray DM. Biomarkers in neonatal hypoxic-ischemic encephalopathy-Review of the literature to date and future directions for research. Handb Clin Neurol 2019; 162: 281-293
• 3 Xu Y, Tian Y, Tian Y, Li X, Zhao P. Autophagy activation involved in hypoxic-ischemic brain injury induces cognitive and memory impairment in neonatal rats. J Neurochem 2016; 139 (05) 795-805
• 4 Zhao F, Qu Y, Liu J. et al. Microarray profiling and co-expression network analysis of LncRNAs and mRNAs in neonatal rats following hypoxic-ischemic brain damage. Sci Rep 2015; 5: 13850
• 5 Dong X, Zhao Y, Huang Y, Yu L, Yang X, Gao F. Analysis of long noncoding RNA expression profiles in the whole blood of neonates with hypoxic-ischemic encephalopathy. J Cell Biochem 2018
• 6 Wang J, Cao B, Han D, Sun M, Feng J. Long non-coding RNA H19 induces cerebral ischemia reperfusion injury via activation of autophagy. Aging Dis 2017; 8 (01) 71-84
• 7 Descloux C, Ginet V, Clarke PG, Puyal J, Truttmann AC. Neuronal death after perinatal cerebral hypoxia-ischemia: focus on autophagy-mediated cell death. Int J Dev Neurosci 2015; 45: 75-85
• 8 Wang S, Xue H, Xu Y, Niu J, Zhao P. Sevoflurane postconditioning inhibits autophagy through activation of the extracellular signal-regulated kinase cascade, alleviating hypoxic-ischemic brain injury in neonatal rats. Neurochem Res 2019; 44 (02) 347-356
• 9 Guo X, Liu Y, Zhao Y. et al. Role of the PI3K–mTOR autophagy pathway in nerve damage in rats with intermittent hypoxia–aggravated whole brain ischemia. Mol Med Rep 2019; 20 (02) 1411-1417
• 10 Balduini W, De Angelis V, Mazzoni E, Cimino M. Simvastatin protects against long-lasting behavioral and morphological consequences of neonatal hypoxic/ischemic brain injury. Stroke 2001; 32 (09) 2185-2191
• 11 Zheng Z, Zhang L, Qu Y. et al. Mesenchymal stem cells protect against hypoxia-ischemia brain damage by enhancing autophagy through brain derived neurotrophic factor/mammalin target of rapamycin signaling pathway. Stem Cells 2018; 36 (07) 1109-1121
• 12 Zhao YH, Ji TF, Luo Q, Yu JL. Long non-coding RNA H19 induces hippocampal neuronal apoptosis via Wnt signaling in a streptozotocin-induced rat model of diabetes mellitus. Oncotarget 2017; 8 (39) 64827-64839
• 13 Tao J, Shen C, Sun Y, Chen W, Yan G. Neuroprotective effects of pinocembrin on ischemia/reperfusion-induced brain injury by inhibiting autophagy. Biomed Pharmacother 2018; 106: 1003-1010
• 14 Tu L, Wang Y, Chen D. et al. Protective effects of notoginsenoside R1 via regulation of the PI3K-Akt-mTOR/JNK pathway in neonatal cerebral hypoxic-ischemic brain injury. Neurochem Res 2018; 43 (06) 1210-1226
• 15 Chen C, Liu M, Tang Y. et al. LncRNA H19 is involved in myocardial ischemic preconditioning via increasing the stability of nucleolin protein. J Cell Physiol 2020; 235 (09) 5985-5994
• 16 Sanchez-Parra C, Jacovetti C, Dumortier O. et al. Contribution of the Long noncoding RNA H19 to β-cell mass expansion in neonatal and adult rodents. Diabetes 2018; 67 (11) 2254-2267
• 17 Li X, Wang H, Yao B, Xu W, Chen J, Zhou X. lncRNA H19/miR-675 axis regulates cardiomyocyte apoptosis by targeting VDAC1 in diabetic cardiomyopathy. Sci Rep 2016; 6: 36340
• 18 Yuan Y, Zheng Z. Geniposide protects PC-12 cells against oxygen and glucose deprivation-induced injury by up-regulation of long-noncoding RNA H19. Life Sci 2019; 216: 176-182
• 19 Zhuo C, Jiang R, Lin X, Shao M. LncRNA H19 inhibits autophagy by epigenetically silencing of DIRAS3 in diabetic cardiomyopathy. Oncotarget 2017; 8 (01) 1429-1437
• 20 Huang TJ, Ren JJ, Zhang QQ. et al. IGFBPrP1 accelerates autophagy and activation of hepatic stellate cells via mutual regulation between H19 and PI3K/AKT/mTOR pathway. Biomed Pharmacother 2019; 116: 109034
• 21 Peng T, Liu X, Wang J. et al. Long noncoding RNA HAGLROS regulates apoptosis and autophagy in Parkinson's disease via regulating miR-100/ATG10 axis and PI3K/Akt/mTOR pathway activation. Artif Cells Nanomed Biotechnol 2019; 47 (01) 2764-2774
• 22 Jacobs SE, Berg M, Hunt R, Tarnow-Mordi WO, Inder TE, Davis PG. Cooling for newborns with hypoxic ischaemic encephalopathy. Cochrane Database Syst Rev 2013; (01) CD003311
• 23 Xiao Z, Qiu Y, Lin Y. et al. Blocking lncRNA H19-miR-19a-Id2 axis attenuates hypoxia/ischemia induced neuronal injury. Aging (Albany NY) 2019; 11 (11) 3585-3600
• 24 Xu LX, Tang XJ, Yang YY. et al. Neuroprotective effects of autophagy inhibition on hippocampal glutamate receptor subunits after hypoxia-ischemia-induced brain damage in newborn rats. Neural Regen Res 2017; 12 (03) 417-424
• 25 Gong LC, Xu HM, Guo GL, Zhang T, Shi JW, Chang C. Long non-coding RNA H19 protects H9c2 cells against hypoxia-induced injury by targeting microRNA-139. Cell Physiol Biochem 2017; 44 (03) 857-869
• 26 Uchiyama Y, Koike M, Shibata M. Autophagic neuron death in neonatal brain ischemia/hypoxia. Autophagy 2008; 4 (04) 404-408
• 27 Zhang X, Cheng L, Xu L. et al. The lncRNA, H19 mediates the protective effect of hypoxia postconditioning against hypoxia-reoxygenation injury to senescent cardiomyocytes by targeting microRNA-29b-3p. Shock 2019; 52 (02) 249-256
• 28 Liu X, Deng Y, Xu Y, Jin W, Li H. MicroRNA-223 protects neonatal rat cardiomyocytes and H9c2 cells from hypoxia-induced apoptosis and excessive autophagy via the Akt/mTOR pathway by targeting PARP-1. J Mol Cell Cardiol 2018; 118: 133-146
• 29 Xue H, Xu Y, Wang S. et al. Sevoflurane post-conditioning alleviates neonatal rat hypoxic-ischemic cerebral injury via Ezh2-regulated autophagy. Drug Des Devel Ther 2019; 13: 1691-1706