Effects of crocin on the enhancement of in vitro neurogenesis: Involvement of Notch and CREB/BDNF signaling pathways

Document Type : Original Article

Authors

1 Department of Anatomical Science, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran

2 Kerman Neuroscience Research Center (KNRC), Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran

3 Afzal Research Institute (NGO), Kerman, Iran

Abstract

Objective(s): Adult neurogenesis, the process of generating new neurons, continues throughout life. Unfortunately, this process is insufficient in pathological conditions and needs to be promoted. Crocin, the active component of saffron, affects neurogenesis in vivo and in vitro. We aimed to investigate the enhancing effects of crocin on the neurogenesis of adipose-derived mesenchymal stem cells in the presence of retinoic acid, as well as the molecular pathways involved. 
Materials and Methods: Differentiation capacities and stemness potential of harvested ADSCs were evaluated by differentiating into osteocytes and adipocytes, and expression of mesenchymal CD markers by flow cytometry. The optimum dose of crocin was assessed with an MTT assay. Crocin, retinoic acid, CREB/BDNF, and Notch inhibitors and their combination were added to the culture medium. Jag1, Hes1, Notch, and BDNF gene expression were analyzed by RT-PCR on days 7, 14, and 21, while CREB, DCX, SOX2, and NeuN expression were analyzed by immunofluorescence.
Results: Expression of mesenchymal CD markers as well as adipogenic and osteogenic differentiation confirmed the origin and properties of ADSCs. The optimal dose of crocin was 1 mM. Crocin significantly (P<0.05) increased, while inhibitors (DATP&Naphthol) significantly (P<0.05) decreased Jag1, Hes1, Notch, and BDNF expression. Immunofluorescent assessments showed that expression of DCX, BDNF, NeuN, and Sox2 proteins increased significantly (P<0.05) after crocin administration and decreased significantly (P<0.05) after inhibitor administration.
Conclusion: Crocin can be used as an enhancer for neural differentiation of MSCs in vitro in the presence of retinoic acid. The mechanism is proposed through Notch and CREB/BDNF signaling pathways.

Keywords

Main Subjects

References

1. Colucci-D’Amato L, Bonavita V, di Porzio U. The end of the central dogma of neurobiology: stem cells and neurogenesis in adult CNS. Neurol Sci 2006; 27: 266-270.
2. Kuhn HG, Eisch AJ, Spalding K, Peterson DA. Detection and Phenotypic Characterization of Adult Neurogenesis. Cold Spring Harb Perspect Biol 2016; 8: a025981.
3. Ernst A, Alkass K, Bernard S, Salehpour M, Perl S, Tisdale J, et al. Neurogenesis in the striatum of the adult human brain. Cell 2014; 156: 1072-1083.
4. Yuan TF, Arias-Carrión O. Adult neurogenesis in the hypothalamus: Evidence, functions, and implications. CNS Neurol Disord Drug Targets 2011; 10: 433-439.
5. Kole AJ, Annis RP, Deshmukh M. Mature neurons: equipped for survival. Cell Death Dis 2013; 4: e689.
6. Yadirgi G, Marino S. Adult neural stem cells and their role in brain pathology. J Pathol 2009; 217: 242-253.
7. Urbán N, Guillemot F. Neurogenesis in the embryonic and adult brain: same regulators, different roles. Front Cell Neurosci 2014; 8: 396.
8. Wang LL, Zhang CL. Engineering new neurons: In vivo reprogramming in mammalian brain and spinal cord. Cell Tissue Res 2018; 371: 201-212.
9. Cameron HA, Glover LR. Adult neurogenesis: beyond learning and memory. Annual Rev. Psychol 2015; 66: 53-81.
10.    Poulose SM, Miller MG, Scott T, Shukitt-Hale B. Nutritional Factors Affecting Adult Neurogenesis and Cognitive Function. Advances in nutrition 2017; 8: 804-811.
11.    Jorgensen C. Adult mammalian neurogenesis and motivated behaviors. Integr Zool 2018; 13: 655-672.
12.    Corona R, Retana-Márquez S, Portillo W, Paredes RG. Sexual behavior increases cell proliferation in the rostral migratory stream and promotes the differentiation of the new cells into neurons in the accessory olfactory bulb of female rats. Front Neurosci 2016; 10: 48.
13.    Glasper ER, Hyer MM, Katakam J, Harper R, Ameri C, Wolz T. Fatherhood contributes to increased hippocampal spine density and anxiety regulation in California mice. Brain Behav 2016; 6: e00416.
14.    Ebrahimi B, Vafaei S, Rastegar-Moghaddam SHR, Hosseini M, Tajik Yabr F, Mohammadipour A. Crocin administration from childhood to adulthood increases hippocampal neurogenesis and synaptogenesis in male mice. J Kerman Univ Med Sci 2021; 28: 243-251.
15.    Pachón-Peña G, Yu G, Tucker A, Wu X, Vendrell J, Bunnell BA, et al. Stromal stem cells from adipose tissue and bone marrow of age-matched female donors display distinct immunophenotypic profiles. J Cell Physiol 2011; 226: 843-851.
16.    Yan Y, Ma T, Gong K, Ao Q, Zhang X, Gong Y. Adipose-derived mesenchymal stem cell transplantation promotes adult neurogenesis in the brains of Alzheimer’s disease mice. Neural Regen Res 2014; 9: 798-805.
17.    Radhakrishnan S, Trentz OA, Reddy MS, Rela M, Kandasamy M, Sellathamby S. In vitro transdifferentiation of human adipose tissue-derived stem cells to neural lineage cells - a stage-specific incidence. Adipocyte 2019; 8: 164-177.
18.    Roballo K, Coelho da Silveira J, Bressan F, Souza A, Mattos Pereira V, Pinzon J, et al. Neurons-derived extracellular vesicles promote neural differentiation of ADSCs: a model to prevent peripheral nerve degeneration. Sci Rep 2019; 9: 112-113.
19.    Mollazadeh H, Emami SA, Hosseinzadeh H. Razi’s Al-Hawi and saffron (Crocus sativus): A review. Iran J Basic Med Sci 2015; 18: 1153-1166.
20.    Assimopoulou AN, Sinakos Z, Papageorgiou VP. Radical scavenging activity of Crocus sativus L. extract and its bioactive constituents. Phytother Res 2005; 19: 997-1000.
21.    Mohamadpour AH, Ayati Z, Parizadeh MR, Rajbai O, Hosseinzadeh H. Safety evaluation of crocin (a constituent of saffron) tablets in healthy volunteers. Iran J Basic Med Sci 2013;16: 39-46.
22.    Haeri P, Mohammadipour A, Heidari Z, Ebrahimzadeh-Bideskan A. Neuroprotective effect of crocin on substantia nigra in MPTP-induced Parkinson’s disease model of mice. Anat Sci Int 2019; 94: 119-127.
23.    Rios J-L, Recio M, Giner R, Máñez S. An update review of saffron and its active constituents. Phytother Res. 1996; 10: 189-193.
24.    Tamaddonfard E, Farshid AA, Asri-Rezaee S, Javadi S, Khosravi V, Rahman B, et al. Crocin improved learning and memory impairments in streptozotocin-induced diabetic rats. Iran J Basic Med Sci 2013; 16: 91-100.
25.    Baghishani F, Mohammadipour A, Hosseinzadeh H, Hosseini M, Ebrahimzadeh-Bideskan A. The effects of tramadol administration on hippocampal cell apoptosis, learning and memory in adult rats and neuroprotective effects of crocin. Metab Brain Dis 2018; 33: 907-916.
26.    Tao W, Ruan J, Wu R, Zhao M, Zhao T, Qi M, et al. A natural carotenoid crocin exerts antidepressant action by promoting adult hippocampal neurogenesis through Wnt/β-catenin signaling. J Adv Res 2022; 43: 219–231.
27.    Li S, Liu X, Lei J, Yang J, Tian P, Gao Y. Crocin Protects podocytes against oxidative stress and inflammation induced by high glucose through inhibition of NF-κB. Cell Physiol Biochem 2017; 42: 1481-1492.
28.    Ebrahimzadeh A, Moghadam SY, Rahimi H, Motaghinejad M, Motevalian M, Safari S, et al. Crocin acts as a neuroprotective mediator against methylphenidate‑induced neurobehavioral and neurochemical sequelae: Possible role of the CREB-BDNF signaling pathway. Acta Neurobiol Exp (Wars) 2019; 79: 352-366.
29.    An B, Ma Y, Xu Y, Liu X, Zhang X, Zhang J, et al. Crocin regulates the proliferation and migration of neural stem cells after cerebral ischemia by activating the Notch1 pathway. Folia Neuropathol. 2020; 58: 201-212.
30.    Takahashi K, Kurokawa K, Miyagawa K, Mochida-Saito A, Nemoto Y, Iwasa H, et al. Antidementia effects of Enterococcus faecalis 2001 are associated with enhancement of hippocampal neurogenesis via the ERK-CREB-BDNF pathway in olfactory bulbectomized mice. Physiol Behav 2020; 223: 112997.
31.    O’Leary OF, Ogbonnaya ES, Felice D, Levone BR, L CC, Fitzgerald P, et al. The vagus nerve modulates BDNF expression and neurogenesis in the hippocampus. Eur Neuropsychopharmacol 2018; 28: 307-316.
32.    Kopan R, Ilagan MX. The canonical Notch signaling pathway: unfolding the activation mechanism. Cell 2009; 137: 216-233.
33.    Borggrefe T, Liefke R. Fine-tuning of the intracellular canonical Notch signaling pathway. Cell Cycle 2012; 11 :264-276.
34.    Ables JL, Decarolis NA, Johnson MA, Rivera PD, Gao Z, Cooper DC, et al. Notch1 is required for maintenance of the reservoir of adult hippocampal stem cells. J Neurosci 2010; 30: 10484-10492.
35.    Jagasia R, Steib K, Englberger E, Herold S, Faus-Kessler T, Saxe M, et al. GABA-cAMP response element-binding protein signaling regulates maturation and survival of newly generated neurons in the adult hippocampus. J Neurosci 2009; 29 :7966-7977.
36.    Vitaliano GD, Kim JK, Kaufman MJ, Adam CW, Zeballos G, Shanmugavadivu A, et al. Clathrin-nanoparticles deliver BDNF to hippocampus and enhance neurogenesis, synaptogenesis and cognition in HIV/neuroAIDS mouse model. Commun Biol 2022; 5: 1-17.
37.    Krawczenko A, Klimczak A. Adipose tissue-derived mesenchymal stem/stromal cells and their contribution to angiogenic processes in tissue regeneration. Int J Mol Sci 2022; 23: 2425.
38.    Salehinejad P, Alitheen NB, Ali AM, Omar AR, Mohit M, Janzamin E, et al. Comparison of different methods for the isolation of mesenchymal stem cells from human umbilical cord Wharton’s jelly. In vitro Cell Dev Biol Anim 2012; 48: 75-83.
39.    Morales-Garcia JA, Calleja-Conde J, Lopez-Moreno JA, Alonso-Gil S, Sanz-SanCristobal M, Riba J, et al. N,N-dimethyltryptamine compound found in the hallucinogenic tea ayahuasca, regulates adult neurogenesis in vitro and in vivo. Transl Psychiatry 2020; 10: 331.
40.    Dong Z, Huo J, Liang A, Chen J, Chen G, Liu D. Gamma-Secretase Inhibitor (DAPT), a potential therapeutic target drug, caused neurotoxicity in planarian regeneration by inhibiting Notch signaling pathway. Sci Total Environ 2021; 781: 146735.
41.    Uttarkar S, Dukare S, Bopp B, Goblirsch M, Jose J, Klempnauer KH. Naphthol AS-E phosphate inhibits the activity of the transcription factor myb by blocking the interaction with the KIX domain of the coactivator p300. Mol Cancer Ther 2015; 14: 1276-1285.
42.    Bejoy J, Bijonowski B, Marzano M, Jeske R, Ma T, Li Y. Wnt-notch signaling interactions during neural and astroglial patterning of human stem cells. Tissue Eng Part A 2020; 26: 419-431.
43.    Li BX, Yamanaka K, Xiao X. Structure–activity relationship studies of naphthol AS-E and its derivatives as anticancer agents by inhibiting CREB-mediated gene transcription. Bioorg Med Chem 2012; 20: 6811-6820.
44.    Li B, Qin K, Wang B, Liu B, Yu W, Li Z, et al. Crocin promotes osteogenesis differentiation of bone marrow mesenchymal stem cells. In Vitro Cell Dev Biol Anim 2020; 56: 680-688.
45.    Saharkhiz M, Razavi FE, Riahi SM, Ayadilord M, Rostami Z, Naseri M. An in vitro study of the effects of crocin on the modulation of DSPP, VEGF-A, HLA-G5, STAT3 and CD200 expression in human dental pulp stem cells. Cell Reprogram 2021; 23: 239-249.
46.    Kim YR, Ahn SM, Pak ME, Lee HJ, Jung DH, Shin Y-I, et al. Potential benefits of mesenchymal stem cells and electroacupuncture on the trophic factors associated with neurogenesis in mice with ischemic stroke. Sci Rep 2018; 8: 20-44.
47.    Cawthorn WP, Scheller EL, MacDougald OAJJolr. Adipose tissue stem cells meet preadipocyte commitment: going back to the future. J Lipid Res 2012; 53: 227-246.
48.    Mazini L, Ezzoubi M, Malka GJScr, therapy. Overview of current adipose-derived stem cell (ADSCs) processing involved in therapeutic advancements: flow chart and regulation updates before and after COVID-19. Stem Cell Res Ther 2021; 12: 1-17.
49.    Ahmadi S, Nabiuni M, Tahmaseb M, Amini EJIJoPRI. Enhanced neural differentiation of epidermal neural crest stem cell by synergistic effect of lithium carbonate and Crocin on BDNF and GDNF expression as neurotrophic factors. Iran J Pharm Res 2021; 20: 95-106.
50.    Grochowski CM, Loomes KM, Spinner NBJG. Jagged1 (JAG1): Structure, expression, and disease associations. Gene 2016; 576: 381-384.
51.    Ohtsuka T, Ishibashi M, Gradwohl G, Nakanishi S, Guillemot F, Kageyama RJTEj. Hes1 and Hes5 as notch effectors in mammalian neuronal differentiation. EMBO J 1999; 18: 2196-2207.
52.    Rezai M, Mahmoodi M, Kaeidi A, Karimabad M, Khoshdel A, Hajizadeh M. Effect of crocin carotenoid on BDNF and CREB gene expression in brain ventral tegmental area of morphine treated rats. Asian Pac J Trop Biomed 2018; 8: 387-393.
53.    Dorri SA, Hosseinzadeh H, Abnous K, Hasani FV, Robati RY, Razavi BM. Involvement of brain-derived neurotrophic factor (BDNF) on malathion induced depressive-like behavior in subacute exposure and protective effects of crocin. Iran J Basic Med Sci 2015; 18: 958-966.
54.    Ahmadi S, Nabiuni M, Tahmaseb M, Amini E. Enhanced neural differentiation of epidermal neural crest stem cell by synergistic effect of lithium carbonate and crocin on BDNF and GDNF expression as neurotrophic factors. Iran J Pharm Res 2021; 20: 95-106.
55.    Tao W, Ruan J, Wu R, Zhao M, Zhao T, Qi M, et al. A natural carotenoid crocin exerts antidepressant action by promoting adult hippocampal neurogenesis through Wnt/β-catenin signaling. J Adv Res 2023; 43: 219-231.
56.    Motaghinejad M, Safari S, Feizipour S, Sadr S. Crocin may be useful to prevent or treatment of alcohol induced neurodegeneration and neurobehavioral sequels via modulation of CREB/BDNF and Akt/GSK signaling pathway. Med Hypotheses. 2019; 124: 21-25.
57.    Asghari N, Irani S, Pezeshki-Moddaress M, Zandi M, Mohamadali M. Neuronal differentiation of mesenchymal stem cells by polyvinyl alcohol/Gelatin/crocin and beta-carotene. Mol Biol Rep 2022; 49: 2999-3006.
58.    Vahdati Hassani F, Naseri V, Razavi BM, Mehri S, Abnous K, Hosseinzadeh HJDJoPS. Antidepressant effects of crocin and its effects on transcript and protein levels of CREB, BDNF, and VGF in rat hippocampus. Daru 2014; 22: 1-9.
59.    Tao W, Ruan J, Wu R, Zhao M, Zhao T, Qi M, et al. A natural carotenoid crocin exerts antidepressant action by promoting adult hippocampal neurogenesis through Wnt/β-catenin signaling. J Adv Res 2023; 43: 219-231.
 
Iranian Journal of Basic Medical Sciences
Volume 27, Issue 7
July 2024
Pages 914-922

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