Long non-coding RNA and mRNA analysis of Ang II-induced neuronal dysfunction
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.
KeywordsLong-chain non-coding RNA MRNA Neurological dysfunction Angiotensin II
- Ang II
Long non-coding RNA
Human nerve cell
Dulbecco’s modified eagle medium: nutrient mixture F-12
Fetal bovine serum
Oxygen consumption rate
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.
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.
- 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. https://doi.org/10.1161/STROKEAHA.108.517284 CrossRefGoogle Scholar
- 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. https://doi.org/10.1161/CIRCULATIONAHA.114.011675 CrossRefGoogle Scholar
- 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. https://doi.org/10.1161/CIRCULATIONAHA.116.026991 CrossRefGoogle Scholar
- 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. https://doi.org/10.1161/HYPERTENSIONAHA.114.04139 CrossRefGoogle Scholar
- 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. https://doi.org/10.1134/S0026898418040109 CrossRefGoogle Scholar
- 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. https://doi.org/10.1016/j.neuroscience.2018.07.026 CrossRefGoogle Scholar
- 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
- 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. https://doi.org/10.3389/fnmol.2017.00433 CrossRefGoogle Scholar
- 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. https://doi.org/10.1007/s12192-016-0724-2 CrossRefGoogle Scholar
- 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. https://doi.org/10.1093/toxsci/kfr163 CrossRefGoogle Scholar