Long non-coding RNA and mRNA analysis of Ang II-induced neuronal dysfunction

  • Lin-Lin Shao
  • Yue-Hua Jiang
  • Ling-Yu Jiang
  • Chuan-Hua YangEmail author
  • Ying-Zi Qi
Original Article


The sustained activation of Angiotensin II (Ang II) induces the remodelling of neurovascular units, inflammation and oxidative stress reactions in the brain. Long non-coding RNAs (lncRNAs) play a crucial regulatory role in the pathogenesis of hypertensive neuronal damage. The present study aimed to substantially extend the list of potential candidate genes involved in Ang II-related neuronal damage. This study assessed apoptosis and energy metabolism with Annexin V/PI staining and a Seahorse assay after Ang II exposure in SH-SY5Y cells. The expression of mRNA and lncRNA was investigated by transcriptome sequencing. The integrated analysis of mRNA and lncRNAs and the molecular mechanism of Ang II on neuronal injury was analysed by bioinformatics. Ang II increased the apoptosis rate and reduced the energy metabolism of SH-SY5Y cells. The data showed that 702 mRNAs and 821 lncRNAs were differentially expressed in response to Ang II exposure (244 mRNAs and 432 lncRNAs were upregulated, 458 mRNAs and 389 lncRNAs were downregulated) (fold change ≥ 1.5, P < 0.05). GO and KEGG analyses showed that both DE mRNA and DE lncRNA were enriched in the metabolism, differentiation, apoptosis and repair of nerve cells. This is the first report of the lncRNA–mRNA integrated profile of SH-SY5Y cells induced by Ang II. The novel targets revealed that the metabolism of the vitamin B group, the synthesis of unsaturated fatty acids and glycosphingolipids are involved in the Ang II-related cognitive impairment. Sphingolipid metabolism, the Hedgehog signalling pathway and vasopressin-regulated water reabsorption play important roles in nerve damage.


Long-chain non-coding RNA MRNA Neurological dysfunction Angiotensin II 


Ang II

Angiotensin II


Long non-coding RNA


Human nerve cell


Dulbecco’s modified eagle medium: nutrient mixture F-12


Fetal bovine serum


Oxygen consumption rate


Differential expression


Fold change


GeNE ontology


Biological process


Cellular components


Molecular function


Angiotensin converting enzyme 2


Low-density lipoprotein receptor-related protein 2


Endothelin-converting enzyme-like 1


Vasoactive intestinal peptide



This study was supported by National Natural Science Foundation of China No. 81573916 and Shandong Province ‘Taishan Scholar’ Construction Project Funds No. 2018-35. We thank Yixin Yin of Shanghai Biotechnology Corporation for technical assistance and constructive suggestions.

Author Contributions

LLS, performed the experiments and drafted the manuscript. YHJ designed the study and performed the experiments. LYJ carried out methodology and data analysis. CHY designed the study, contributed to discussion and carried out important revisions of the article. YZQ contributed to data curation and software applications. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Dunn KM, Nelson MT (2014) Neurovascular signaling in the brain and the pathological consequences of hypertension. Am J Physiol Heart Circ Physiol 306:H1–H14. CrossRefGoogle Scholar
  2. 2.
    Yamamoto E, Tamamaki N, Nakamura T, Kataoka K, Tokutomi Y, Dong YF, Fukuda M, Matsuba S, Ogawa H, Kim-Mitsuyama S (2008) Excess salt causes cerebral neuronal apoptosis and inflammation in stroke-prone hypertensive rats through angiotensin II-induced NADPH oxidase activation. Stroke 39:3049–3056. CrossRefGoogle Scholar
  3. 3.
    De Bundel D, Smolders I, Vanderheyden P, Michotte Y (2008) Ang II and Ang IV: unraveling the mechanism of action on synaptic plasticity, memory, and epilepsy. CNS Neurosci Ther 14:315–339. CrossRefGoogle Scholar
  4. 4.
    Marques FZ, Booth SA, Charchar FJ (2015) The emerging role of non-coding RNA in essential hypertension and blood pressure regulation. J Hum Hypertens 29:459–467. CrossRefGoogle Scholar
  5. 5.
    Murakami K (2015) Non-coding RNAs and hypertension-unveiling unexpected mechanisms of hypertension by the dark matter of the genome. Curr Hypertens Rev 11:80–90CrossRefGoogle Scholar
  6. 6.
    Wu G, Cai J, Han Y, Chen J, Huang ZP, Chen C, Cai Y, Huang H, Yang Y, Liu Y, Xu Z, He D, Zhang X, Hu X, Pinello L, Zhong D, He F, Yuan GC, Wang DZ, Zeng C (2014) LincRNA-p21 regulates neointima formation, vascular smooth muscle cell proliferation, apoptosis, and atherosclerosis by enhancing p53 activity. Circulation 130:1452–1465. CrossRefGoogle Scholar
  7. 7.
    Leisegang MS, Fork C, Josipovic I, Richter FM, Preussner J, Hu J, Miller MJ, Epah J, Hofmann P, Gunther S, Moll F, Valasarajan C, Heidler J, Ponomareva Y, Freiman TM, Maegdefessel L, Plate KH, Mittelbronn M, Uchida S, Kunne C, Stellos K, Schermuly RT, Weissmann N, Devraj K, Wittig I, Boon RA, Dimmeler S, Pullamsetti SS, Looso M, Miller FJ Jr, Brandes RP (2017) Long noncoding RNA MANTIS facilitates endothelial angiogenic function. Circulation 136:65–79. CrossRefGoogle Scholar
  8. 8.
    Leung A, Trac C, Jin W, Lanting L, Akbany A, Saetrom P, Schones DE, Natarajan R (2013) Novel long noncoding RNAs are regulated by angiotensin II in vascular smooth muscle cells. Circ Res 113:266–278. CrossRefGoogle Scholar
  9. 9.
    Wharton W, Goldstein FC, Zhao L, Steenland K, Levey AI, Hajjar I (2015) Modulation of renin-angiotensin system may slow conversion from mild cognitive impairment to Alzheimer’s disease. J Am Geriatr Soc 63:1749–1756. CrossRefGoogle Scholar
  10. 10.
    Rygiel K (2016) Can angiotensin-converting enzyme inhibitors impact cognitive decline in early stages of Alzheimer’s disease? An overview of research evidence in the elderly patient population. J Postgrad Med 62:242–248. CrossRefGoogle Scholar
  11. 11.
    Tadic M, Cuspidi C, Hering D (2016) Hypertension and cognitive dysfunction in elderly: blood pressure management for this global burden. BMC Cardiovasc Disord 16:208. CrossRefGoogle Scholar
  12. 12.
    Cifuentes D, Poittevin M, Dere E, Broqueres-You D, Bonnin P, Benessiano J, Pocard M, Mariani J, Kubis N, Merkulova-Rainon T, Levy BI (2015) Hypertension accelerates the progression of Alzheimer-like pathology in a mouse model of the disease. Hypertension 65:218–224. CrossRefGoogle Scholar
  13. 13.
    Kozin SA, Polshakov VI, Mezentsev YV, Ivanov AS, Zhokhov SS, Yurinskaya MM, Vinokurov MG, Makarov AA, Mitkevich VA (2018) Enalaprilat inhibits zinc-dependent oligomerization of metal-binding domain of amyloid-beta isoforms and protects human neuroblastoma cells from toxic action of these isoforms. Mol Biol 52:683–691. CrossRefGoogle Scholar
  14. 14.
    Shah SA, Yoon GH, Chung SS, Abid MN, Kim TH, Lee HY, Kim MO (2017) Novel osmotin inhibits SREBP2 via the AdipoR1/AMPK/SIRT1 pathway to improve Alzheimer’s disease neuropathological deficits. Mol Psychiatry 22:407–416. CrossRefGoogle Scholar
  15. 15.
    Mastroeni D, Nolz J, Khdour OM, Sekar S, Delvaux E, Cuyugan L, Liang WS, Hecht SM, Coleman PD (2018) Oligomeric amyloid beta preferentially targets neuronal and not glial mitochondrial-encoded mRNAs. Alzheimers Dement 14:775–786. CrossRefGoogle Scholar
  16. 16.
    Wanka H, Lutze P, Staar D, Grunow B, Peters BS, Peters J (2018) An alternative renin isoform is cardioprotective by modulating mitochondrial metabolism. J Cell Mol Med 22:5991–6001. CrossRefGoogle Scholar
  17. 17.
    Brand MD, Nicholls DG (2011) Assessing mitochondrial dysfunction in cells. Biochem J 435:297–312. CrossRefGoogle Scholar
  18. 18.
    Chun HJ, Lee Y, Kim AH, Lee J (2016) Methylglyoxal causes cell death in neural progenitor cells and impairs adult hippocampal neurogenesis. Neurotox Res 29:419–431. CrossRefGoogle Scholar
  19. 19.
    Kushwah N, Jain V, Dheer A, Kumar R, Prasad D, Khan N (2018) Hypobaric hypoxia-induced learning and memory impairment: elucidating the role of small conductance Ca(2+)-Activated K(+) channels. Neuroscience 388:418–429. CrossRefGoogle Scholar
  20. 20.
    Xia H, Suda S, Bindom S, Feng Y, Gurley SB, Seth D, Navar LG, Lazartigues E (2011) ACE2-mediated reduction of oxidative stress in the central nervous system is associated with improvement of autonomic function. PLoS ONE 6:e22682. CrossRefGoogle Scholar
  21. 21.
    Landowski LM, Pavez M, Brown LS, Gasperini R, Taylor BV, West AK, Foa L (2016) Low-density lipoprotein receptor-related proteins in a novel mechanism of axon guidance and peripheral nerve regeneration. J Biol Chem 291:1092–1102. CrossRefGoogle Scholar
  22. 22.
    Nagata K, Kiryu-Seo S, Tamada H, Okuyama-Uchimura F, Kiyama H, Saido TC (2016) ECEL1 mutation implicates impaired axonal arborization of motor nerves in the pathogenesis of distal arthrogryposis. Acta Neuropathol 132:111–126. CrossRefGoogle Scholar
  23. 23.
    Sragovich S, Merenlender-Wagner A, Gozes I (2017) ADNP Plays a key role in autophagy: from autism to Schizophrenia and Alzheimer’s disease. Bioessays. Google Scholar
  24. 24.
    Kochanowski J, Uchman D, Litwiniuk A, Kalisz M, Wolinska-Witort E, Martynska L, Baranowska B, Bik W (2015) Assessment of plasma brain-derived neurotrophic factor (BDNF), activity-dependent neurotrophin protein (ADNP) and vasoactive intestinal peptide (VIP) concentrations in treatment-naive humans with multiple sclerosis. Neuro Endocrinol Lett 36:148–152Google Scholar
  25. 25.
    Kasacka I, Piotrowska Z, Janiuk I (2015) Influence of renovascular hypertension on the distribution of vasoactive intestinal peptide in the stomach and heart of rats. Exp Biol Med 240:1402–1407. CrossRefGoogle Scholar
  26. 26.
    Aureli M, Grassi S, Prioni S, Sonnino S, Prinetti A (2015) Lipid membrane domains in the brain. Biochim Biophys Acta 1851:1006–1016. CrossRefGoogle Scholar
  27. 27.
    Aureli M, Samarani M, Loberto N, Bassi R, Murdica V, Prioni S, Prinetti A, Sonnino S (2014) The glycosphingolipid hydrolases in the central nervous system. Mol Neurobiol 50:76–87. CrossRefGoogle Scholar
  28. 28.
    Henriques A, Croixmarie V, Bouscary A, Mosbach A, Keime C, Boursier-Neyret C, Walter B, Spedding M, Loeffler JP (2017) Sphingolipid metabolism is dysregulated at transcriptomic and metabolic levels in the spinal cord of an animal model of amyotrophic lateral sclerosis. Front Mol Neurosci 10:433. CrossRefGoogle Scholar
  29. 29.
    Ferent J, Traiffort E (2015) Hedgehog: multiple paths for multiple roles in shaping the brain and spinal cord. Neuroscientist 21:356–371. CrossRefGoogle Scholar
  30. 30.
    Prager-Khoutorsky M, Choe KY, Levi DI, Bourque CW (2017) Role of vasopressin in rat models of salt-dependent hypertension. Curr Hypertens Rep 19:42. CrossRefGoogle Scholar
  31. 31.
    Li CY, Zhang L, Li J, Qi CL, Li DY, Liu X, Qu X (2017) Effect of endogenous arginine-vasopressin arising from the paraventricular nucleus on learning and memory functions in vascular dementia model rats. Biomed Res Int 2017:3214918. Google Scholar
  32. 32.
    Singh KK, Matkar PN, Pan Y, Quan A, Gupta V, Teoh H, Al-Omran M, Verma S (2017) Endothelial long non-coding RNAs regulated by oxidized LDL. Mol Cell Biochem 431:139–149. CrossRefGoogle Scholar
  33. 33.
    Ehret GB, O’Connor AA, Weder A, Cooper RS, Chakravarti A (2009) Follow-up of a major linkage peak on chromosome 1 reveals suggestive QTLs associated with essential hypertension: GenNet study. Eur J Hum Genet 17:1650–1657. CrossRefGoogle Scholar
  34. 34.
    Kamide K, Kokubo Y, Yang J, Tanaka C, Hanada H, Takiuchi S, Inamoto N, Banno M, Kawano Y, Okayama A, Tomoike H, Miyata T (2005) Hypertension susceptibility genes on chromosome 2p24-p25 in a general Japanese population. J Hypertens 23:955–960CrossRefGoogle Scholar
  35. 35.
    Vidan-Jeras B, Gregoric A, Jurca B, Jeras M, Bohinjec M (2000) Possible influence of genes located on chromosome 6 within or near to the major histocompatibility complex on development of essential hypertension. Pflugers Arch 439:R60–R62CrossRefGoogle Scholar
  36. 36.
    Knight J, Munroe PB, Pembroke JC, Caulfield MJ (2003) Human chromosome 17 in essential hypertension. Ann Hum Genet 67:193–206CrossRefGoogle Scholar
  37. 37.
    Shorbagi S, Brown IR (2016) Dynamics of the association of heat shock protein HSPA6 (Hsp70B’) and HSPA1A (Hsp70-1) with stress-sensitive cytoplasmic and nuclear structures in differentiated human neuronal cells. Cell Stress Chaperones 21:993–1003. CrossRefGoogle Scholar
  38. 38.
    Song D, Nishiyama M, Kimura S (2016) Potent inhibition of angiotensin AT1 receptor signaling by RGS8: importance of the C-terminal third exon part of its RGS domain. J Recept Signal Transduct Res 36:478–487. CrossRefGoogle Scholar
  39. 39.
    Del Barrio L, Martin-de-Saavedra MD, Romero A, Parada E, Egea J, Avila J, McIntosh JM, Wonnacott S, Lopez MG (2011) Neurotoxicity induced by okadaic acid in the human neuroblastoma SH-SY5Y line can be differentially prevented by alpha7 and beta2* nicotinic stimulation. Toxicol Sci 123:193–205. CrossRefGoogle Scholar
  40. 40.
    Parkkila S, Parkkila AK, Rajaniemi H, Shah GN, Grubb JH, Waheed A, Sly WS (2001) Expression of membrane-associated carbonic anhydrase XIV on neurons and axons in mouse and human brain. Proc Natl Acad Sci USA 98:1918–1923. CrossRefGoogle Scholar
  41. 41.
    Altun I, Kurutas EB (2016) Vitamin B complex and vitamin B12 levels after peripheral nerve injury. Neural Regen Res 11:842–845. CrossRefGoogle Scholar
  42. 42.
    Bazinet RP, Laye S (2014) Polyunsaturated fatty acids and their metabolites in brain function and disease. Nat Rev Neurosci 15:771–785. CrossRefGoogle Scholar
  43. 43.
    Schnaar RL, Gerardy-Schahn R, Hildebrandt H (2014) Sialic acids in the brain: gangliosides and polysialic acid in nervous system development, stability, disease, and regeneration. Physiol Rev 94:461–518. CrossRefGoogle Scholar
  44. 44.
    Jackson L, Eldahshan W, Fagan SC, Ergul A (2018) Within the brain: the renin angiotensin system. Int J Mol Sci 1:9. Google Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Lin-Lin Shao
    • 1
  • Yue-Hua Jiang
    • 2
  • Ling-Yu Jiang
    • 2
  • Chuan-Hua Yang
    • 2
    Email author
  • Ying-Zi Qi
    • 1
  1. 1.First Clinical Medical CollegeShandong University of Traditional Chinese MedicineJinanChina
  2. 2.Department of CardiovascularAffiliated Hospital of Shandong University of Traditional Chinese MedicineJinanChina

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