The Inhibitory Effects of Hesperetin, a Natural Flavonoid, against Substance P-induced Inflammatory Responses in Human Primary Glioblastoma Cells
Abstract
Objectives: As a key inflammatory neuropeptide, substance p (SP), is involved in Glioblastoma multiforme (GBM) carcinogenesis and tumor progression. Thus, the proinflammatory effects of SP must be strictly regulated in GBM patients. The purpose of the study is to examine whether Hesperetin, a natural flavonoid found in citrus fruits with strong anti-inflammatory and anticancer effects, can regulate SP-induced inflammation in GBM primary human cells.
Methods: The primary human cells were derived from fresh surgically resected tissue samples of GBM patients and characterized by immunocytochemical analysis of Ki-67. MTT assay was used to evaluate cell proliferation. Apoptosis was detected by an Annexin-V/propidium iodide assay kit using flow cytometry. The levels of the specific inflammatory mediators are measured by western blotting and enzyme-linked immunosorbent assay.
Results: We observed that Hesperetin effectively reduced GBM cell viability in a dose-dependent manner which was associated with the induction of apoptosis. Obtained findings indicated SP significantly increased the protein expression of phosphorylated-NF-κB, as the main regulator of inflammatory processes, and the pro-inflammatory cytokines (IL-1β, TNF-α) while Hesperetin treatment reduced the effects of SP.
Conclusion: Taken together, our findings highlight the role of SP-induced inflammatory responses in GBM pathology and suggest that Hesperetin could be an effective therapeutic strategy in attenuating SP-associated inflammation.
1. Hanif F, Muzaffar K, Perveen K, Malhi SM, Simjee SU. Glioblastoma multiforme: a review of its epidemiology and pathogenesis through clinical presentation and treatment. Asian Pacific journal of cancer prevention: APJCP. 2017;18(1):3.
2. Grochans S, Cybulska AM, Simińska D, Korbecki J, Kojder K, Chlubek D, Baranowska-Bosiacka I. Epidemiology of glioblastoma multiforme–literature review. Cancers. 2022;14(10):2412.
3. Jain KK. A Critical Overview of Targeted Therapies for Glioblastoma. Front Oncol. 2018;8:419.
4. Wu W, Klockow JL, Zhang M, Lafortune F, Chang E, Jin L, et al. Glioblastoma multiforme (GBM): An overview of current therapies and mechanisms of resistance. Pharmacol Res. 2021;171:105780.
5. Gupta T, Sarin R. Poor-prognosis high-grade gliomas: evolving an evidence-based standard of care. Lancet Oncol. 2002;3(9):557-64.
6. Ulasov I, Singh V, Laevskaya A, Timashev P, Kharwar RK. Inflammatory Mediators and GBM Malignancy: Current Scenario and Future Prospective. Discov Med. 2023;35(177):458-75.
7. Johnson MB, Young AD, Marriott I. The Therapeutic Potential of Targeting Substance P/NK-1R Interactions in Inflammatory CNS Disorders. Front Cell Neurosci. 2016;10:296.
8. Sun J, Ramnath RD, Zhi L, Tamizhselvi R, Bhatia M. Substance P enhances NF-kappaB transactivation and chemokine response in murine macrophages via ERK1/2 and p38 MAPK signaling pathways. Am J Physiol Cell Physiol. 2008;294(6):C1586-96.
9. Kowalski ML, Sliwinska-Kowalska M, Kaliner MA. Neurogenic inflammation, vascular permeability, and mast cells. II. Additional evidence indicating that mast cells are not involved in neurogenic inflammation. J Immunol. 1990;145(4):1214-21.
10. Palma C, Bigioni M, Irrissuto C, Nardelli F, Maggi C, Manzini S. Anti-tumour activity of tachykinin NK1 receptor antagonists on human glioma U373 MG xenograft. British Journal of Cancer. 2000;82(2):480-7.
11. Hennig IM, Laissue JA, Horisberger U, Reubi JC. Substance‐P receptors in human primary neoplasms: Tumoral and vascular localization. International Journal of Cancer. 1995;61(6):786-92.
12. Akazawa T, Kwatra SG, Goldsmith LE, Richardson MD, Cox EA, Sampson JH, Kwatra MM. A constitutively active form of neurokinin 1 receptor and neurokinin 1 receptor-mediated apoptosis in glioblastomas. J Neurochem. 2009;109(4):1079-86.
13. Garg A, Garg S, Zaneveld LJ, Singla AK. Chemistry and pharmacology of the Citrus bioflavonoid hesperidin. Phytother Res. 2001;15(8):655-69.
14. Tejada S, Pinya S, Martorell M, Capo X, Tur JA, Pons A, Sureda A. Potential Anti-inflammatory Effects of Hesperidin from the Genus Citrus. Curr Med Chem. 2018;25(37):4929-45.
15. Iranshahi M, Rezaee R, Parhiz H, Roohbakhsh A, Soltani F. Protective effects of flavonoids against microbes and toxins: The cases of hesperidin and hesperetin. Life Sci. 2015;137:125-32.
16. Aswar M, Kute P, Mahajan S, Mahajan U, Nerurkar G, Aswar U. Protective effect of hesperetin in rat model of partial sciatic nerve ligation induced painful neuropathic pain: an evidence of anti-inflammatory and anti-oxidative activity. Pharmacol Biochem Behav. 2014;124:101-7.
17. Jin Q, Zhang W, Qiu XG, Yan W, You G, Liu YW, et al. Gene expression profiling reveals Ki-67 associated proliferation signature in human glioblastoma. Chin Med J (Engl). 2011;124(17):2584-8.
18. Raychaudhuri B, Han Y, Lu T, Vogelbaum MA. Aberrant constitutive activation of nuclear factor kappaB in glioblastoma multiforme drives invasive phenotype. J Neurooncol. 2007;85(1):39-47.
19. O'Connor TM, O'Connell J, O'Brien DI, Goode T, Bredin CP, Shanahan F. The role of substance P in inflammatory disease. J Cell Physiol. 2004;201(2):167-80.
20. Rosso M, Munoz M, Berger M. The role of neurokinin-1 receptor in the microenvironment of inflammation and cancer. ScientificWorldJournal. 2012;2012:381434.
21. Esteban F, Ramos-Garcia P, Munoz M, Gonzalez-Moles MA. Substance P and Neurokinin 1 Receptor in Chronic Inflammation and Cancer of the Head and Neck: A Review of the Literature. Int J Environ Res Public Health. 2021;19(1).
22. Gillespie E, Leeman SE, Watts LA, Coukos JA, O'Brien MJ, Cerda SR, et al. Truncated neurokinin-1 receptor is increased in colonic epithelial cells from patients with colitis-associated cancer. Proceedings of the National Academy of Sciences. 2011;108(42):17420-5.
23. Landskron G, De la Fuente M, Thuwajit P, Thuwajit C, Hermoso MA. Chronic inflammation and cytokines in the tumor microenvironment. J Immunol Res. 2014;2014:149185.
24. Kore RA, Abraham EC. Inflammatory cytokines, interleukin-1 beta and tumor necrosis factor-alpha, upregulated in glioblastoma multiforme, raise the levels of CRYAB in exosomes secreted by U373 glioma cells. Biochemical and biophysical research communications. 2014;453(3):326-31.
25. Godwin P, Baird AM, Heavey S, Barr MP, O'Byrne KJ, Gately K. Targeting nuclear factor-kappa B to overcome resistance to chemotherapy. Front Oncol. 2013;3:120.
26. Singh S, Kumaravel S, Dhole S, Roy S, Pavan V, Chakraborty S. Neuropeptide Substance P Enhances Inflammation-Mediated Tumor Signaling Pathways and Migration and Proliferation of Head and Neck Cancers. Indian J Surg Oncol. 2021;12(Suppl 1):93-102.
27. Lieb K, Fiebich BL, Berger M, Bauer J, Schulze-Osthoff K. The neuropeptide substance P activates transcription factor NF-kappa B and kappa B-dependent gene expression in human astrocytoma cells. J Immunol. 1997;159(10):4952-8.
28. Dehelean CA, Marcovici I, Soica C, Mioc M, Coricovac D, Iurciuc S, et al. Plant-derived anticancer compounds as new perspectives in drug discovery and alternative therapy. Molecules. 2021;26(4):1109.
29. Tsai WH, Wu CH, Yu HJ, Chien CT. l-Theanine inhibits proinflammatory PKC/ERK/ICAM-1/IL-33 signaling, apoptosis, and autophagy formation in substance P-induced hyperactive bladder in rats. Neurourol Urodyn. 2017;36(2):297-307.
30. Li PC, Li SC, Lin YJ, Liang JT, Chien CT, Shaw CF. Thoracic vagal efferent nerve stimulation evokes substance P-induced early airway bronchonstriction and late proinflammatory and oxidative injury in the rat respiratory tract. J Biomed Sci. 2005;12(4):671-81.
31. Li PC, Chen WC, Chang LC, Lin SC. Substance P acts via the neurokinin receptor 1 to elicit bronchoconstriction, oxidative stress, and upregulated ICAM-1 expression after oil smoke exposure. Am J Physiol Lung Cell Mol Physiol. 2008;294(5):L912-20.
32. Li Q, Miao Z, Wang R, Yang J, Zhang D. Hesperetin induces apoptosis in human glioblastoma cells via p38 MAPK activation. Nutrition and Cancer. 2020;72(3):538-45.
33. Cheng Q, Mao L, Huang H, Tang L, Jiang H, Zhang Y, Mu Q. Hesperetin ameliorates glioblastoma by inhibiting proliferation, inducing apoptosis, and suppressing metastasis. Transl Cancer Res. 2022;11(6):1781-94.
2. Grochans S, Cybulska AM, Simińska D, Korbecki J, Kojder K, Chlubek D, Baranowska-Bosiacka I. Epidemiology of glioblastoma multiforme–literature review. Cancers. 2022;14(10):2412.
3. Jain KK. A Critical Overview of Targeted Therapies for Glioblastoma. Front Oncol. 2018;8:419.
4. Wu W, Klockow JL, Zhang M, Lafortune F, Chang E, Jin L, et al. Glioblastoma multiforme (GBM): An overview of current therapies and mechanisms of resistance. Pharmacol Res. 2021;171:105780.
5. Gupta T, Sarin R. Poor-prognosis high-grade gliomas: evolving an evidence-based standard of care. Lancet Oncol. 2002;3(9):557-64.
6. Ulasov I, Singh V, Laevskaya A, Timashev P, Kharwar RK. Inflammatory Mediators and GBM Malignancy: Current Scenario and Future Prospective. Discov Med. 2023;35(177):458-75.
7. Johnson MB, Young AD, Marriott I. The Therapeutic Potential of Targeting Substance P/NK-1R Interactions in Inflammatory CNS Disorders. Front Cell Neurosci. 2016;10:296.
8. Sun J, Ramnath RD, Zhi L, Tamizhselvi R, Bhatia M. Substance P enhances NF-kappaB transactivation and chemokine response in murine macrophages via ERK1/2 and p38 MAPK signaling pathways. Am J Physiol Cell Physiol. 2008;294(6):C1586-96.
9. Kowalski ML, Sliwinska-Kowalska M, Kaliner MA. Neurogenic inflammation, vascular permeability, and mast cells. II. Additional evidence indicating that mast cells are not involved in neurogenic inflammation. J Immunol. 1990;145(4):1214-21.
10. Palma C, Bigioni M, Irrissuto C, Nardelli F, Maggi C, Manzini S. Anti-tumour activity of tachykinin NK1 receptor antagonists on human glioma U373 MG xenograft. British Journal of Cancer. 2000;82(2):480-7.
11. Hennig IM, Laissue JA, Horisberger U, Reubi JC. Substance‐P receptors in human primary neoplasms: Tumoral and vascular localization. International Journal of Cancer. 1995;61(6):786-92.
12. Akazawa T, Kwatra SG, Goldsmith LE, Richardson MD, Cox EA, Sampson JH, Kwatra MM. A constitutively active form of neurokinin 1 receptor and neurokinin 1 receptor-mediated apoptosis in glioblastomas. J Neurochem. 2009;109(4):1079-86.
13. Garg A, Garg S, Zaneveld LJ, Singla AK. Chemistry and pharmacology of the Citrus bioflavonoid hesperidin. Phytother Res. 2001;15(8):655-69.
14. Tejada S, Pinya S, Martorell M, Capo X, Tur JA, Pons A, Sureda A. Potential Anti-inflammatory Effects of Hesperidin from the Genus Citrus. Curr Med Chem. 2018;25(37):4929-45.
15. Iranshahi M, Rezaee R, Parhiz H, Roohbakhsh A, Soltani F. Protective effects of flavonoids against microbes and toxins: The cases of hesperidin and hesperetin. Life Sci. 2015;137:125-32.
16. Aswar M, Kute P, Mahajan S, Mahajan U, Nerurkar G, Aswar U. Protective effect of hesperetin in rat model of partial sciatic nerve ligation induced painful neuropathic pain: an evidence of anti-inflammatory and anti-oxidative activity. Pharmacol Biochem Behav. 2014;124:101-7.
17. Jin Q, Zhang W, Qiu XG, Yan W, You G, Liu YW, et al. Gene expression profiling reveals Ki-67 associated proliferation signature in human glioblastoma. Chin Med J (Engl). 2011;124(17):2584-8.
18. Raychaudhuri B, Han Y, Lu T, Vogelbaum MA. Aberrant constitutive activation of nuclear factor kappaB in glioblastoma multiforme drives invasive phenotype. J Neurooncol. 2007;85(1):39-47.
19. O'Connor TM, O'Connell J, O'Brien DI, Goode T, Bredin CP, Shanahan F. The role of substance P in inflammatory disease. J Cell Physiol. 2004;201(2):167-80.
20. Rosso M, Munoz M, Berger M. The role of neurokinin-1 receptor in the microenvironment of inflammation and cancer. ScientificWorldJournal. 2012;2012:381434.
21. Esteban F, Ramos-Garcia P, Munoz M, Gonzalez-Moles MA. Substance P and Neurokinin 1 Receptor in Chronic Inflammation and Cancer of the Head and Neck: A Review of the Literature. Int J Environ Res Public Health. 2021;19(1).
22. Gillespie E, Leeman SE, Watts LA, Coukos JA, O'Brien MJ, Cerda SR, et al. Truncated neurokinin-1 receptor is increased in colonic epithelial cells from patients with colitis-associated cancer. Proceedings of the National Academy of Sciences. 2011;108(42):17420-5.
23. Landskron G, De la Fuente M, Thuwajit P, Thuwajit C, Hermoso MA. Chronic inflammation and cytokines in the tumor microenvironment. J Immunol Res. 2014;2014:149185.
24. Kore RA, Abraham EC. Inflammatory cytokines, interleukin-1 beta and tumor necrosis factor-alpha, upregulated in glioblastoma multiforme, raise the levels of CRYAB in exosomes secreted by U373 glioma cells. Biochemical and biophysical research communications. 2014;453(3):326-31.
25. Godwin P, Baird AM, Heavey S, Barr MP, O'Byrne KJ, Gately K. Targeting nuclear factor-kappa B to overcome resistance to chemotherapy. Front Oncol. 2013;3:120.
26. Singh S, Kumaravel S, Dhole S, Roy S, Pavan V, Chakraborty S. Neuropeptide Substance P Enhances Inflammation-Mediated Tumor Signaling Pathways and Migration and Proliferation of Head and Neck Cancers. Indian J Surg Oncol. 2021;12(Suppl 1):93-102.
27. Lieb K, Fiebich BL, Berger M, Bauer J, Schulze-Osthoff K. The neuropeptide substance P activates transcription factor NF-kappa B and kappa B-dependent gene expression in human astrocytoma cells. J Immunol. 1997;159(10):4952-8.
28. Dehelean CA, Marcovici I, Soica C, Mioc M, Coricovac D, Iurciuc S, et al. Plant-derived anticancer compounds as new perspectives in drug discovery and alternative therapy. Molecules. 2021;26(4):1109.
29. Tsai WH, Wu CH, Yu HJ, Chien CT. l-Theanine inhibits proinflammatory PKC/ERK/ICAM-1/IL-33 signaling, apoptosis, and autophagy formation in substance P-induced hyperactive bladder in rats. Neurourol Urodyn. 2017;36(2):297-307.
30. Li PC, Li SC, Lin YJ, Liang JT, Chien CT, Shaw CF. Thoracic vagal efferent nerve stimulation evokes substance P-induced early airway bronchonstriction and late proinflammatory and oxidative injury in the rat respiratory tract. J Biomed Sci. 2005;12(4):671-81.
31. Li PC, Chen WC, Chang LC, Lin SC. Substance P acts via the neurokinin receptor 1 to elicit bronchoconstriction, oxidative stress, and upregulated ICAM-1 expression after oil smoke exposure. Am J Physiol Lung Cell Mol Physiol. 2008;294(5):L912-20.
32. Li Q, Miao Z, Wang R, Yang J, Zhang D. Hesperetin induces apoptosis in human glioblastoma cells via p38 MAPK activation. Nutrition and Cancer. 2020;72(3):538-45.
33. Cheng Q, Mao L, Huang H, Tang L, Jiang H, Zhang Y, Mu Q. Hesperetin ameliorates glioblastoma by inhibiting proliferation, inducing apoptosis, and suppressing metastasis. Transl Cancer Res. 2022;11(6):1781-94.
| Files | ||
| Issue | Vol 2 No 2 (2024) | |
| Section | Original Articles | |
| DOI | https://doi.org/10.18502/abi.v2i2.17938 | |
| Keywords | ||
| substance p Hesperetin Glioblastoma inflammation NF-κB | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Aliee A, Hashemy SI, Ebrahimi S. The Inhibitory Effects of Hesperetin, a Natural Flavonoid, against Substance P-induced Inflammatory Responses in Human Primary Glioblastoma Cells. ABI. 2024;2(2):103-110.
