Go to Home

Search

-Advanced Search   -Search Guidelines

Korean J Physiol Pharmacol.  2024 Jul;28(4):361-377. 10.4196/kjpp.2024.28.4.361.

Analysis of the mechanism of fibrauretine alleviating Alzheimer's disease based on transcriptomics and proteomics

Affiliations
  • 1College of Chinese Medicinal Materials, Jilin Agricultural University, Changchun 130118, China
  • 2Key Laboratory of Animal Production, Product Quality and Security, Ministry of Education of China, Changchun 130118, China
  • 3Jilin Provincial Engineering Research Center for Efficient Breeding and Product Development of Sika Deer of China, Changchun 130118, China

Abstract

The dried rattan stem of the Fibraurea Recisa Pierre plant contains the active ingredient known as fibrauretine (FN). Although it greatly affects Alzheimer's disease (AD), the mechanism of their effects still remains unclear. Proteomics and transcriptomics analysis methods were used in this study to determine the mechanism of FN in the treatment of AD. AD model is used through bilateral hippocampal injection of Aβ1-40. After successful modeling, FN was given for 30 days. The results showed that FN could improve the cognitive dysfunction of AD model rats, reduce the expression of Aβ and P-Tau, increase the content of acetylcholine and reduce the activity of acetylcholinesterase. The Kyoto Encyclopedia of Genes and Genomes enriched differentially expressed genes and proteins are involved in signaling pathways including metabolic pathway, AD, pathway in cancer, PI3K-AKT signaling pathway, and cAMP signaling pathway. Transcriptomics and proteomics sequencing resulted in 19 differentially expressed genes and proteins. Finally, in contrast to the model group, after FN treatment, the protein expressions and genes associated with the PI3K-AKT pathway were significantly improved in RT-qPCR and Western blot and assays. This is consistent with the findings of transcriptomic and proteomic analyses. Our study found that, FN may improve some symptoms of AD model rats through PI3K-AKT signaling pathway.

Keyword

Alzheimer's disease; fibrauretine; Proteomics; Transcriptomics

Figure

  • Fig. 1 The experimental process of analyzing the mechanism of FN improving AD symptoms based on transcriptomics and proteomics. FN, fibrauretine; AD, Alzheimer’s disease.

  • Fig. 2 Result of Morris water maze experiment. (A) Escape latency. (B) Time of crossing platform. (C) Thermal infrared trajectories of positioning navigation. (D) Thermal infrared trajectories of space exploration. Values are presented as mean ± SD. N = 10. FN, fibrauretine. ##p < 0.01 vs. control group. **p < 0.01, *p < 0.05 vs. model group.

  • Fig. 3 The effects of FN on the contents of A β and P-Tau in hippocampus. (A) The expression of Aβ and P-Tau was detected by immunohistochemistry (200×). (B) Average optical density of Aβ. (C) Average optical density of P-Tau. Values are presented as mean ± SD. N = 6. FN, fibrauretine; Aβ, amyloid β. ##p < 0.01 vs. control group. **p < 0.01, vs. model group.

  • Fig. 4 Differentially expressed genes between groups. (A) Volcano plot of differentially expressed genes between the control and model group. (B) Volcano plot of differentially expressed genes between the model and FN group. (C) Venn of common differentially expressed genes in the control group and model group, model group and FN group. (D) Differentially expressed genes clustering heatmap. N = 3. FN, fibrauretine.

  • Fig. 5 GO enrichment analysis of differentially expressed genes. (A) Bubble plots of the enrichment analysis of differentially expressed genes in biological processes between the control and model group. (B) Bubble plots of the enrichment analysis of differentially expressed genes in cell composition between the control and model group. (C) Bubble plots of the enrichment analysis of differentially expressed genes in molecular function between the control and model group. (D) Bubble plots of the enrichment analysis of differentially expressed genes in biological processes between the model and FN group. (E) Bubble plots of the enrichment analysis of differentially expressed genes in cell composition between the model and FN group. (F) Bubble plots of the enrichment analysis of differentially expressed genes in molecular function between the model and FN group. N = 3. GO, Gene Ontology; FN, fibrauretine.

  • Fig. 6 KEGG enrichment analysis. (A) KEGG classification annotation map of differentially expressed genes between the control group and model group. (B) KEGG classification annotation map of differentially expressed genes between the model group and FN group. (C) Top 20 pathways enriched to KEGG in the control group vs. model group. (D) Top 20 pathways enriched to KEGG in the model group vs. FN group. N = 3. KEGG, Kyoto Encyclopedia of Genes and Genomes; FN, fibrauretine.

  • Fig. 7 The mRNA expression level of SPP1, Nr4a1, KLF4 in each group was detected by RT-qPCR. Values are presented as mean ± SD. N = 6. FN, fibrauretine. ##p < 0.01 vs. control group. **p < 0.01, *p < 0.05 vs. model group.

  • Fig. 8 PCA analysis and differentially expressed proteins volcano map, Wayne map and heat map. (A) PCA analysis between groups. (B) Differentially expressed proteins volcano map between the control group and model group. (C) Venn of common differentially expressed proteins in the control group and model group, model group and FN group. (D) Differentially expressed proteins volcanoes between the model group and FN group. (E) Differentially expressed proteins clustering heat map. N = 3. PCA, principal component analysis; FN, fibrauretine.

  • Fig. 9 GO enrichment analysis of differentially expressed proteins. (A) Control group vs. model group in terms of molecular function enriched to entries. (B) Control group vs. model group enriched to entries on biological processes. (C) Entries enriched to cell composition in the control group vs. model group. (D) The entries enriched in molecular function in the model group vs. FN group. (E) Entries enriched to biological processes in the model group vs. FN group. (F) Entries enriched to cell composition in the model group vs. FN group. N = 3. GO, Gene Ontology; FN, fibrauretine.

  • Fig. 10 KEGG classification of differentially expressed proteins in each group. (A) KEGG classification of differentially expressed proteins between the control group and model group. (B) KEGG classification of differentially expressed proteins between the model group and FN group. N = 3. KEGG, Kyoto Encyclopedia of Genes and Genomes; FN, fibrauretine.

  • Fig. 11 The effect of FN on the expression of PI3K-AKT pathway-related proteins. (A) Western blot was used to detect the expression of PI3K-AKT pathway-related proteins. (B) The level of PI3K. (C) The level of AKT. (D) The level of MTOR. Values are presented as mean ± SD. N = 3. FN, fibrauretine. ##p < 0.01, #p < 0.05 vs. control group. *p < 0.05 vs. model group.

  • Fig. 12 Overlapping of differentially expressed protein-coding genes and differentially expressed genes. N = 3.


Reference

1. Scheltens P, De Strooper B, Kivipelto M, Holstege H, Chételat G, Teunissen CE, Cummings J, van der Flier WM. 2021; Alzheimer's disease. Lancet. 397:1577–1590. DOI: 10.1016/S0140-6736(20)32205-4. PMID: 33667416.
2. Dujardin P, Vandenbroucke RE, Van Hoecke L. 2022; Fighting fire with fire: the immune system might be key in our fight against Alzheimer's disease. Drug Discov Today. 27:1261–1283. DOI: 10.1016/j.drudis.2022.01.004. PMID: 35032668.
3. Ye JY, Li L, Hao QM, Qin Y, Ma CS. 2020; β-Sitosterol treatment attenuates cognitive deficits and prevents amyloid plaque deposition in amyloid protein precursor/presenilin 1 mice. Korean J Physiol Pharmacol. 24:39–46. DOI: 10.4196/kjpp.2020.24.1.39. PMID: 31908573. PMCID: PMC6940499.
4. Xing Z, He Z, Wang S, Yan Y, Zhu H, Gao Y, Zhao Y, Zhang L. 2018; Ameliorative effects and possible molecular mechanisms of action of fibrauretine from Fibraurea recisa Pierre on d-galactose/AlCl3-mediated Alzheimer's disease. RSC Adv. 8:31646–31657. DOI: 10.1039/C8RA05356A. PMID: 35548215. PMCID: PMC9085853.
5. Wang Y, Lin Y, Wang L, Zhan H, Luo X, Zeng Y, Wu W, Zhang X, Wang F. 2020; TREM2 ameliorates neuroinflammatory response and cognitive impairment via PI3K/AKT/FoxO3a signaling pathway in Alzheimer's disease mice. Aging (Albany NY). 12:20862–20879. DOI: 10.18632/aging.104104. PMID: 33065553. PMCID: PMC7655179.
6. Sebastian Monasor L, Müller SA, Colombo AV, Tanrioever G, König J, Roth S, Liesz A, Berghofer A, Piechotta A, Prestel M, Saito T, Saido TC, Herms J, Willem M, Haass C, Lichtenthaler SF, Tahirovic S. 2020; Fibrillar Aβ triggers microglial proteome alterations and dysfunction in Alzheimer mouse models. Elife. 9:e54083. DOI: 10.7554/eLife.54083. PMID: 32510331. PMCID: PMC7279888.
7. Sur B, Lee B. 2022; Myricetin prevents sleep deprivation-induced cognitive impairment and neuroinflammation in rat brain via regulation of brain-derived neurotropic factor. Korean J Physiol Pharmacol. 26:415–425. DOI: 10.4196/kjpp.2022.26.6.415. PMID: 36302617. PMCID: PMC9614391.
8. Celik Topkara K, Kilinc E, Cetinkaya A, Saylan A, Demir S. 2022; Therapeutic effects of carvacrol on beta-amyloid-induced impairments in in vitro and in vivo models of Alzheimer's disease. Eur J Neurosci. 56:5714–5726. DOI: 10.1111/ejn.15565. PMID: 34904309.
9. Lee B, Yeom M, Shim I, Lee H, Hahm DH. 2020; Inhibitory effect of carvacrol on lipopolysaccharide-induced memory impairment in rats. Korean J Physiol Pharmacol. 24:27–37. DOI: 10.4196/kjpp.2020.24.1.27. PMID: 31908572. PMCID: PMC6940503.
10. Kirino T. 1982; Delayed neuronal death in the gerbil hippocampus following ischemia. Brain Res. 239:57–69. DOI: 10.1016/0006-8993(82)90833-2. PMID: 7093691.
11. Shukla PK, Meena AS, Manda B, Gomes-Solecki M, Dietrich P, Dragatsis I, Rao R. 2018; Lactobacillus plantarum prevents and mitigates alcohol-induced disruption of colonic epithelial tight junctions, endotoxemia, and liver damage by an EGF receptor-dependent mechanism. FASEB J. 32:fj201800351R. DOI: 10.1096/fj.201800351R. PMID: 29912589. PMCID: PMC6181630.
12. Yang X, Guan Y, Yan B, Xie Y, Zhou M, Wu Y, Yao L, Qiu X, Yan F, Chen Y, Huang L. 2021; Evidence-based complementary and alternative medicine bioinformatics approach through network pharmacology and molecular docking to determine the molecular mechanisms of Erjing pill in Alzheimer's disease. Exp Ther Med. 22:1252. DOI: 10.3892/etm.2021.10687. PMID: 34539848. PMCID: PMC8438686.
13. Park GW, Hwang H, Kim KH, Lee JY, Lee HK, Park JY, Ji ES, Park SR, Yates JR 3rd, Kwon KH, Park YM, Lee HJ, Paik YK, Kim JY, Yoo JS. 2016; Integrated proteomic pipeline using multiple search engines for a proteogenomic study with a controlled protein false discovery rate. J Proteome Res. 15:4082–4090. DOI: 10.1021/acs.jproteome.6b00376. PMID: 27537616.
14. Bloom GS. 2014; Amyloid-β and tau: the trigger and bullet in Alzheimer disease pathogenesis. JAMA Neurol. 71:505–508. DOI: 10.1001/jamaneurol.2013.5847. PMID: 24493463.
15. Furukawa A, Kawamoto Y, Chiba Y, Takei S, Hasegawa-Ishii S, Kawamura N, Yoshikawa K, Hosokawa M, Oikawa S, Kato M, Shimada A. 2011; Proteomic identification of hippocampal proteins vulnerable to oxidative stress in excitotoxin-induced acute neuronal injury. Neurobiol Dis. 43:706–714. DOI: 10.1016/j.nbd.2011.05.024. PMID: 21669285.
16. Pedrero-Prieto CM, García-Carpintero S, Frontiñán-Rubio J, Llanos-González E, Aguilera García C, Alcaín FJ, Lindberg I, Durán-Prado M, Peinado JR, Rabanal-Ruiz Y. 2020; A comprehensive systematic review of CSF proteins and peptides that define Alzheimer's disease. Clin Proteomics. 17:21. DOI: 10.1186/s12014-020-09276-9. PMID: 32518535. PMCID: PMC7273668.
17. Tsuboyama M, Iqbal MA. 2021; CHL1 deletion is associated with cognitive and language disabilities - Case report and review of literature. Mol Genet Genomic Med. 9:e1725. DOI: 10.1002/mgg3.1725. PMID: 34056867. PMCID: PMC8372067.
18. Zhuang X, Zhang G, Bao M, Jiang G, Wang H, Li S, Wang Z, Sun X. 2023; Development of a novel immune infiltration-related diagnostic model for Alzheimer's disease using bioinformatic strategies. Front Immunol. 14:1147501. DOI: 10.3389/fimmu.2023.1147501. PMID: 37545529. PMCID: PMC10400274.
19. Maes M, DeVos N, Wauters A, Demedts P, Maurits VW, Neels H, Bosmans E, Altamura C, Lin A, Song C, Vandenbroucke M, Scharpe S. 1999; Inflammatory markers in younger vs elderly normal volunteers and in patients with Alzheimer's disease. J Psychiatr Res. 33:397–405. DOI: 10.1016/S0022-3956(99)00016-3. PMID: 10504008.
20. Bai Y, Li L, Dong B, Ma W, Chen H, Yu Y. 2022; Phosphorylation-mediated PI3K-Art signalling pathway as a therapeutic mechanism in the hydrogen-induced alleviation of brain injury in septic mice. J Cell Mol Med. 26:5713–5727. DOI: 10.1111/jcmm.17568. PMID: 36308410. PMCID: PMC9667523.
21. Lissanu Deribe Y. 2016; Interplay between PREX2 mutations and the PI3K pathway and its effect on epigenetic regulation of gene expression in NRAS-mutant melanoma. Small GTPases. 7:178–185. DOI: 10.1080/21541248.2016.1178366. PMID: 27111337. PMCID: PMC5003543.
22. Zhang Q, Li L, Lai Y, Zhao T. 2020; Silencing of SPP1 suppresses progression of tongue cancer by mediating the PI3K/Akt signaling pathway. Technol Cancer Res Treat. 19:1533033820971306. DOI: 10.1177/1533033820971306. PMID: 33174521. PMCID: PMC7672768.
23. Zhang Y, Li S, Cui X, Wang Y. 2023; microRNA-944 inhibits breast cancer cell proliferation and promotes cell apoptosis by reducing SPP1 through inactivating the PI3K/Akt pathway. Apoptosis. 28:1546–1563. DOI: 10.1007/s10495-023-01870-0. PMID: 37486406.
24. Briggs R, Kennelly SP, O'Neill D. 2016; Drug treatments in Alzheimer's disease. Clin Med (Lond). 16:247–253. DOI: 10.7861/clinmedicine.16-3-247. PMID: 27251914. PMCID: PMC5922703.
25. Jia W, Su Q, Cheng Q, Peng Q, Qiao A, Luo X, Zhang J, Wang Y. 2021; Neuroprotective effects of palmatine via the enhancement of antioxidant defense and small heat shock protein expression in Aβ-transgenic Caenorhabditis elegans. Oxid Med Cell Longev. 2021:9966223. DOI: 10.1155/2021/9966223. PMID: 34567416. PMCID: PMC8460366.
26. Pei H, Zeng J, He Z, Zong Y, Zhao Y, Li J, Chen W, Du R. 2023; Palmatine ameliorates LPS-induced HT-22 cells and mouse models of depression by regulating apoptosis and oxidative stress. J Biochem Mol Toxicol. 37:e23225. DOI: 10.1002/jbt.23225. PMID: 36169195.
27. Millan MJ. 2017; Linking deregulation of non-coding RNA to the core pathophysiology of Alzheimer's disease: an integrative review. Prog Neurobiol. 156:1–68. DOI: 10.1016/j.pneurobio.2017.03.004. PMID: 28322921.
28. Verheijen J, Sleegers K. 2018; Understanding Alzheimer disease at the interface between genetics and transcriptomics. Trends Genet. 34:434–447. DOI: 10.1016/j.tig.2018.02.007. PMID: 29573818.
29. Zhang T, Shen Y, Guo Y, Yao J. 2021; Identification of key transcriptome biomarkers based on a vital gene module associated with pathological changes in Alzheimer's disease. Aging (Albany NY). 13:14940–14967. DOI: 10.18632/aging.203017. PMID: 34031265. PMCID: PMC8221319.
30. De Schepper S, Ge JZ, Crowley G, Ferreira LSS, Garceau D, Toomey CE, Sokolova D, Rueda-Carrasco J, Shin SH, Kim JS, Childs T, Lashley T, Burden JJ, Sasner M, Sala Frigerio C, Jung S, Hong S. 2023; Perivascular cells induce microglial phagocytic states and synaptic engulfment via SPP1 in mouse models of Alzheimer's disease. Nat Neurosci. 26:406–415. DOI: 10.1038/s41593-023-01257-z. PMID: 36747024. PMCID: PMC9991912.
31. Zhang Y, Zhu H, Liu L, Ma H, Shi Q, Li D, Ju X. 2023; Three-dimensional culture of rat ovarian granulosa cells shows increased SPP1 and FGF7 expression through the PI3K/Akt pathway. Cell Biol Int. 47:1004–1016. DOI: 10.1002/cbin.11998. PMID: 36701359.
32. Zhang Z, Yu J. 2018; NR4A1 promotes cerebral ischemia reperfusion injury by repressing Mfn2-mediated mitophagy and inactivating the MAPK-ERK-CREB signaling pathway. Neurochem Res. 43:1963–1977. DOI: 10.1007/s11064-018-2618-4. PMID: 30136162.
33. McMorrow JP, Murphy EP. 2011; Inflammation: a role for NR4A orphan nuclear receptors? Biochem Soc Trans. 39:688–693. DOI: 10.1042/BST0390688. PMID: 21428963.
34. Paillasse MR, de Medina P. 2015; The NR4A nuclear receptors as potential targets for anti-aging interventions. Med Hypotheses. 84:135–140. DOI: 10.1016/j.mehy.2014.12.003. PMID: 25543265.
35. Blacher E, Tsai C, Litichevskiy L, Shipony Z, Iweka CA, Schneider KM, Chuluun B, Heller HC, Menon V, Thaiss CA, Andreasson KI. 2022; Aging disrupts circadian gene regulation and function in macrophages. Nat Immunol. 23:229–236. DOI: 10.1038/s41590-021-01083-0. PMID: 34949832. PMCID: PMC9704320.
36. Ji CY, Yuan MX, Chen LJ, Gao HQ, Zhang T, Yin Q. 2022; miR92a represses the viability and migration of nerve cells in Hirschsprung's disease by regulating the KLF4/PI3K/AKT pathway. Acta Neurobiol Exp (Wars). 82:336–346. DOI: 10.55782/ane-2022-032. PMID: 36214716.
37. Andreev VP, Petyuk VA, Brewer HM, Karpievitch YV, Xie F, Clarke J, Camp D, Smith RD, Lieberman AP, Albin RL, Nawaz Z, El Hokayem J, Myers AJ. 2012; Label-free quantitative LC-MS proteomics of Alzheimer's disease and normally aged human brains. J Proteome Res. 11:3053–3067. DOI: 10.1021/pr3001546. PMID: 22559202. PMCID: PMC3445701.
38. Hui W, Feng Y, Ruihua X, Jiongjie H, Dongan C, Yan S, Shidong Z. 2020; Comparative proteomics analysis indicates that palmatine contributes to transepithelial migration by regulating cellular adhesion. Pharm Biol. 58:646–654. Erratum in: Pharm Biol. 2021;59:546. DOI: 10.1080/13880209.2020.1784961. PMID: 32658562. PMCID: PMC7470081.
39. Johnson ECB, Dammer EB, Duong DM, Ping L, Zhou M, Yin L, Higginbotham LA, Guajardo A, White B, Troncoso JC, Thambisetty M, Montine TJ, Lee EB, Trojanowski JQ, Beach TG, Reiman EM, Haroutunian V, Wang M, Schadt E, Zhang B, et al. 2020; Large-scale proteomic analysis of Alzheimer's disease brain and cerebrospinal fluid reveals early changes in energy metabolism associated with microglia and astrocyte activation. Nat Med. 26:769–780. DOI: 10.1038/s41591-020-0815-6. PMID: 32284590. PMCID: PMC7405761.
40. Zhang S, Sun P, Xiao X, Hu Y, Qian Y, Zhang Q. 2022; MicroRNA-21 promotes epithelial-mesenchymal transition and migration of human bronchial epithelial cells by targeting poly (ADP-ribose) polymerase-1 and activating PI3K/AKT signaling. Korean J Physiol Pharmacol. 26:239–253. DOI: 10.4196/kjpp.2022.26.4.239. PMID: 35766002. PMCID: PMC9247709.
41. Long HZ, Cheng Y, Zhou ZW, Luo HY, Wen DD, Gao LC. 2021; PI3K/AKT signal pathway: a target of natural products in the prevention and treatment of Alzheimer's disease and Parkinson's disease. Front Pharmacol. 12:648636. DOI: 10.3389/fphar.2021.648636. PMID: 33935751. PMCID: PMC8082498.
Full Text Links
  • KJPP
Actions
Cited
CITED
export Copy
Close
Share
  • Twitter
  • Facebook
Similar articles

Cited

Download

Close

Save citations to file

Download

Close

Email citations

Send Email
recaptcha 영역

Close

Copyright © 2024 by Korean Association of Medical Journal Editors. All rights reserved.     E-mail: koreamed@kamje.or.kr