Repeated isoflurane exposures of neonatal rats contribute to cognitive dysfunction in juvenile animals: the role of miR-497 in isoflurane-induced neurotoxicity
Abstract
Introduction. Isoflurane anesthesia at the period of brain development can lead to neurotoxicity and long-term cognitive impairment. This study aimed to investigate the role of miR-497 on isoflurane-induced neurotoxicity. Material and methods. Neonatal rats (P7) were subject to isoflurane for 2 h at P7, P9, and P11. MiR-497 and neuron apoptosis were evaluated in hippocampal tissue by qRT-PCR and western blot. Fear conditioning test and Morris water maze were performed to determine cognitive function. The cell viability of isolated hippocampal neuronal cells exposed to isoflurane was measured using MTT test. The regulation of phospholipase D1 (PLD1) by miR-497 in isolated hippocampal neuronal cells was evaluated by luciferase reporter assays and western blot. Immunohistochemistry and TUNEL staining were employed to examine the PLD1 expression and neuronal cell apoptosis in hippocampus of neonatal rats, respectively. Results. Repeated isoflurane anesthesia led to neurons’ apoptosis and long-term cognitive impairment. Isoflurane exposure led to apoptosis and viability reduction in hippocampal neuronal cells. MiR-497 was observed to be upregulated after isoflurane exposure both in vivo and in vitro. Knockdown of miR-497 attenuated isoflurane-induced neuronal cells apoptosis and viability reduction. Furthermore, PLD1 was predicted and then validated as a novel target of miR-497. miR-497 could negatively regulate PLD1 by binding to its 3’-untranslated region. Downregulation of PLD1 was also observed after isoflurane exposure in neonatal rat hippocampus and hippocampal primary neuronal cell cultures. Conclusions. Induction of miR-497 was involved in isoflurane anesthesia-induced cognitive impairment and neuronal cell apoptosis by targeting PLD1. miR-497 may be a novel potential mechanism in isoflurane-induced neurotoxicity so that our findings provide new insight into a better and understanding of the clinical application of isoflurane.
Keywords: isofluraneneurotoxicityrat pupshippocampal neuronsmiR-497phospholipase D1cognitive impairmentqPCRwestern blot
References
- Stratmann G, Sall JW, May LDV, et al. Isoflurane differentially affects neurogenesis and long-term neurocognitive function in 60-day-old and 7-day-old rats. Anesthesiology. 2009; 110(4): 834–848.
- Flick RP, Katusic SK, Colligan RC, et al. Cognitive and behavioral outcomes after early exposure to anesthesia and surgery. Pediatrics. 2011; 128(5): e1053–e1061.
- Sprung J, Flick RP, Wilder RT, et al. Anesthesia for cesarean delivery and learning disabilities in a population-based birth cohort. Anesthesiology. 2009; 111(2): 302–310.
- Lu LX, Yon JH, Carter LB, et al. General anesthesia activates BDNF-dependent neuroapoptosis in the developing rat brain. Apoptosis. 2006; 11(9): 1603–1615.
- Creeley CE. From Drug-Induced Developmental Neuroapoptosis to Pediatric Anesthetic Neurotoxicity-Where Are We Now? Brain Sci. 2016; 6(3).
- Cabrera OH, O'Connor SD, Swiney BS, et al. Caffeine combined with sedative/anesthetic drugs triggers widespread neuroapoptosis in a mouse model of prematurity. J Matern Fetal Neonatal Med. 2017; 30(22): 2734–2741.
- Stratmann G, May LDV, Sall JW, et al. Effect of hypercarbia and isoflurane on brain cell death and neurocognitive dysfunction in 7-day-old rats. Anesthesiology. 2009; 110(4): 849–861.
- Yang S, Gu C, Mandeville ET, et al. Anesthesia and Surgery Impair Blood-Brain Barrier and Cognitive Function in Mice. Front Immunol. 2017; 8: 902.
- Yu D, Li L, Yuan W. Neonatal anesthetic neurotoxicity: Insight into the molecular mechanisms of long-term neurocognitive deficits. Biomed Pharmacother. 2017; 87: 196–199.
- Clausen NG, Hansen TG, Disma N. Anesthesia Neurotoxicity in the Developing Brain: Basic Studies Relevant for Neonatal or Perinatal Medicine. Clin Perinatol. 2019; 46(4): 647–656.
- Piscopo P, Lacorte E, Feligioni M, et al. MicroRNAs and mild cognitive impairment: A systematic review. Ageing Res Rev. 2019; 50: 131–141.
- Sun E, Shi Y. MicroRNAs: Small molecules with big roles in neurodevelopment and diseases. Exp Neurol. 2015; 268: 46–53.
- Issler O, Chen A. Determining the role of microRNAs in psychiatric disorders. Nat Rev Neurosci. 2015; 16(4): 201–212.
- Mai H, Fan W, Wang Y, et al. Intranasal Administration of miR-146a Agomir Rescued the Pathological Process and Cognitive Impairment in an AD Mouse Model. Mol Ther Nucleic Acids. 2019; 18: 681–695.
- Goto G, Hori Y, Ishikawa M, et al. Changes in the gene expression levels of microRNAs in the rat hippocampus by sevoflurane and propofol anesthesia. Mol Med Rep. 2014; 9(5): 1715–1722.
- Zhang N, Ye W, Wang T, et al. Up-regulation of miR-106a targets LIMK1 and contributes to cognitive impairment induced by isoflurane anesthesia in mice. Genes Genomics. 2020; 42(4): 405–412.
- Yan H, Xu T, Zhao H, et al. Isoflurane increases neuronal cell death vulnerability by downregulating miR-214. PLoS One. 2013; 8(2): e55276.
- Lu X, Lv S, Mi Y, et al. Neuroprotective effect of miR-665 against sevoflurane anesthesia-induced cognitive dysfunction in rats through PI3K/Akt signaling pathway by targeting insulin-like growth factor 2. Am J Transl Res. 2017; 9(3): 1344–1356.
- Itesako T, Seki N, Yoshino H, et al. The microRNA expression signature of bladder cancer by deep sequencing: the functional significance of the miR-195/497 cluster. PLoS One. 2014; 9(2): e84311.
- Yin KJ, Deng Z, Huang H, et al. miR-497 regulates neuronal death in mouse brain after transient focal cerebral ischemia. Neurobiol Dis. 2010; 38(1): 17–26.
- Yadav S, Pandey A, Shukla A, et al. miR-497 and miR-302b regulate ethanol-induced neuronal cell death through BCL2 protein and cyclin D2. J Biol Chem. 2011; 286(43): 37347–37357.
- Murphy KL, McGaughy J, Croxson PL, et al. Long-term effects of neonatal single or multiple isoflurane exposures on spatial memory in rats. Front Neurol. 2013; 4(1): 87–95.
- Lin D, Zuo Z. Isoflurane induces hippocampal cell injury and cognitive impairments in adult rats. Neuropharmacology. 2011; 61(8): 1354–1359.
- Yan J, Huang Y, Lu Yi, et al. Repeated administration of ketamine can induce hippocampal neurodegeneration and long-term cognitive impairment via the ROS/HIF-1α pathway in developing rats. Cell Physiol Biochem. 2014; 33(6): 1715–1732.
- Yon JH, Daniel-Johnson J, Carter LB, et al. Anesthesia induces neuronal cell death in the developing rat brain via the intrinsic and extrinsic apoptotic pathways. Neuroscience. 2005; 135(3): 815–827.
- Li XM, Zhou MT, Wang XM, et al. Resveratrol pretreatment attenuates the isoflurane-induced cognitive impairment through its anti-inflammation and -apoptosis actions in aged mice. J Mol Neurosci. 2014; 52(2): 286–293.
- Kong F, Chen S, Cheng Y, et al. Minocycline attenuates cognitive impairment induced by isoflurane anesthesia in aged rats. PLoS One. 2013; 8(4): e61385.
- Zhong T, Guo Q, Zou W, et al. Neonatal isoflurane exposure induces neurocognitive impairment and abnormal hippocampal histone acetylation in mice. PLoS One. 2015; 10(4): e0125815.
- Chai D, Jiang H, Li Q. Isoflurane neurotoxicity involves activation of hypoxia inducible factor-1α via intracellular calcium in neonatal rodents. Brain Res. 2016; 1653: 39–50.
- Pang X, Zhang P, Zhou Y, et al. Dexmedetomidine pretreatment attenuates isoflurane-induced neurotoxicity via inhibiting the TLR2/NF-κB signaling pathway in neonatal rats. Exp Mol Pathol. 2020; 112: 104328.
- Xie C, Wang H, Zhang Yu, et al. Neuroprotective effects of miR-142-5p downregulation against isoflurane-induced neurological impairment. Diagn Pathol. 2020; 15(1): 70.
- Fan D, Yang S, Han Y, et al. Isoflurane-induced expression of miR-140-5p aggravates neurotoxicity in diabetic rats by targeting SNX12. J Toxicol Sci. 2020; 45(2): 69–76.
- Burkhardt U, Stegner D, Hattingen E, et al. Impaired brain development and reduced cognitive function in phospholipase D-deficient mice. Neurosci Lett. 2014; 572: 48–52.
- Ammar MR, Thahouly T, Hanauer A, et al. PLD1 participates in BDNF-induced signalling in cortical neurons. Sci Rep. 2015; 5: 14778.
- Luo LD, Li G, Wang Y. PLD1 promotes dendritic spine development by inhibiting ADAM10-mediated N-cadherin cleavage. Scientific Reports. 2017; 7(1).
- Ammar MR, Humeau Y, Hanauer A, et al. The Coffin-Lowry syndrome-associated protein RSK2 regulates neurite outgrowth through phosphorylation of phospholipase D1 (PLD1) and synthesis of phosphatidic acid. J Neurosci. 2013; 33(50): 19470–19479.
- Liu T, Wang F, LePochat P, et al. Cofilin-mediated Neuronal Apoptosis via p53 Translocation and PLD1 Regulation. Sci Rep. 2017; 7(1): 11532.
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