The Potential of Moringa Leaf Extract to Prevent Aging Targeted Cellular Senescence
Keywords:
Cellular senescence, Molringa Oleifera, p21, ROS, TP53
Abstract
High exposure to ultraviolet light and chemicals causes cell senescence due to the overproduction of reactive oxygen species (ROS). Cell senescence is known as one of the causes of degenerative diseases. Reducing ROS levels using antioxidant compounds can overcome cell senescence. This study aimed to evaluate the potential of Moringa leaf extract (EDK) as an anti-aging agent and predict the molecular mechanisms underlying its activity. EDK active compounds were identified through qualitative phytochemical screening, a cytotoxic assay was evaluate using MTT assay, ROS level calculate under DCFDA flow cytometry assay, and cell senescence analyze using SA-β-galactosidase assay. The molecular mechanism that plays an important role in senescence was analyzed through bioinformatics studies and molecular docking in silico to investigate the interaction strength of the active compound EDK with the target protein. The results showed that the EDK ethanol extract contained flavonoid, phenolic, alkaloid, saponin, and steroid compounds. This extract did not give any cytotoxic effect on NIH3T3 cells, as evidenced by the IC50 value of 420μg/mL. EDK concentrations of ¼IC50 (105μg/mL) and ½IC50 (210μg/mL) significantly reduced ROS level in dose-dependent manner in the NIH3T3 cell population with high oxidative stress induced by Doxorubicin 10nM (Dox). The percentage of senescence cells also decreased due to EDK 210μg/mL administration up to 47.60% compared to the positive aging control Dox 10nM (78.90%). Bioinformatics studies found that p21 and TP53 proteins were most directly affected by EDK active compounds in the senescence pathway. The quercetin compound in EDK has the most robust interaction strength against p21 and TP53 proteins compared to ligands. In conclusion, it can be concluded that the active compound EDK can suppress intracellular ROS levels and compete with p21 and TP53 proteins in preventing cell senescence.
References
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Mijit M, Caracciolo V, Melillo A, Amicarelli F, Giordano A (2020) Role of p53 in the regulation of cellular senescence. Biomolecules. 10(3):1–16
Nouman W, Anwar F, Gull T, Newton A, Rosa E, Domínguez-Perles R (2016) Profiling of polyphenolics, nutrients and antioxidant potential of germplasm’s leaves from seven cultivars of Moringa oleifera lam. Industrial Crops and Products. 83:166–176
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Odusote JK, Owalude DO, Olusegun SJ, Yahya RA (2016) Inhibition efficiency of Moringa oleifera leaf extract on the corrosion of reinforced steel bar in HCl solution. West Indian Journal of Engineering. 38(2):64
Oladeji SO, Adelowo FE, Odelade KA (2016) Mass spectroscopic and phytochemical screening of phenolic compounds in the leaf extract of Senna alata (L.) Roxb. (Fabales: Fabaceae). Brazilian Journal of Biological Sciences. 3(5):209-219
Pantsar T, Poso A (2018) Binding affinity via docking: fact and fiction. Molecules. 23(8):1899
Pedersen JK, Engholm G, Skytthe A, Christensen, K (2016) Cancer and aging: epidemiology and methodological challenges. Acta Oncologica. 55(1):7–12
Pulakat L, Chen HH (2020). Pro-Senescence and anti-senescence mechanisms of cardiovascular aging: cardiac MicroRNA regulation of longevity drug-induced autophagy. Frontiers in Oncology. 11:1–9
Qi L, Zhou Y, Li W, Zheng M, Zhong R, Jin X, Lin Y (2019) Effect of moringa oleifera stem extract on hydrogen peroxide-induced opacity of cultured mouse lens. BMC Complementary and Alternative Medicine. 19(1):1-9
Romanov VS, Pospelov VA, Pospelova TV (2012) Cyclin-dependent kinase inhibitor p21Waf1: contemporary view on its role in senescence and oncogenesis. Biochemistry. 77(6):575–584
Salmon AB (2015) About-face on the metabolic side effects of rapamycin. Oncotarget. 6(5):2585–2586
Schosserer M, Grillari J, Breitenbach M (2017) The dual role of cellular senescence in developing tumors and their response to cancer therapy. Frontiers in Oncology. 7:278
Shtutman M, Chang BD, Schools GP, Broude EV (2017) Cellular model of p21-induced senescence. Methods in Molecular Biology. 1534:31–39
Szklarczyk D, Gable AL, Lyon D, Junge A, Wyder S, Huerta-Cepas J, Simonovic M, Doncheva NT, Morris JH, Bork P, Jensen LJ, Von Mering C (2019) STRING v11: protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets. Nucleid Acid Research. 47(D1): D607–D613
Vrancheva R, Ivanov I, Dincheva I, Badjakov I, Pavlov A (2021) Triterpenoids and other non-polar compounds in leaves of wild and cultivated Vaccinium species. Plants. 10(1):1–16
Wyld L, Bellantuono I, Tchkonia T, Morgan J, Turner O, Foss F, George J, Danson S, Kirkland JL (2020) Senescence and cancer: A review of clinical implications of senescence and senotherapies. Cancers (Basel). 12(8):1–20
Yan C, Xu Z, Huang W (2021) Cellular senescence affects cardiac regeneration and repair in ischemic heart disease. Aging and Disease. 12(2):552–569
Yong-Bing X, Gui-Lin C, Ming-Quan G (2019) Antioxidant and anti-inflammatory activities of the crude extracts of Moringa oleifera from kenya and their correlations with flavonoids. Antioxidants. 8(8):296
Zullaikah S, Naulina RY, Meinawati P, Fauziyah K, Rachimoellah M, Rachmaniah O, Nurkhamidah S, Suari NMIP, Prasetyo EN (2019) Enhanced extraction of phenolic compounds from Moringa oleifera leaves using subcritical water ethanol mixture. IOP Conference Series: Materials Science and Engineering. 543(1):1-7
Bagheri G, Martorell M, Ramizer-Alarcon K, Salehi B, Sharifi-Rad J (2020) View of phytochemical screening of Moringa oleifera leaf extracts and their antimicrobial activities. Cellular and Molecular Biology. 66(1): 20-26
Berrouet C, Dorilas N, Rejniak KA, Tuncer N (2020) Comparison of drug inhibitory effects (IC50) in monolayer and spheroid cultures. Bulletin of Mathematical Biology. 82:1-23
Biran A, Zada L, Abou KP, Vadai E, Roitman L, Ovadya Y, Porat Z, Krizhanovsky V (2017) Quantitative identification of senescent cells in aging and disease. Aging Cell. 16:661–671
Brand RM, Wipf P, Durham A, Epperly MW, Greenberger JS, Falo LD (2018) Targeting mitochondrial oxidative stress to mitigate UV-induced skin damage. Frontiers in Pharmacology. 9:1–10
Chen Q, Sun X, Luo X, Wang J, Hu J, Feng Y (2020) PIK3R3 inhibits cell senescence through p53/p21 signaling. Cell Death and Disease. 11(9):1-12
Choi JB, Kim JH, Lee H, Pak JN, Shim BS, Kim SH (2018) Reactive oxygen species and p53 mediated activation of p38 and caspases is critically involved in kaempferol induced apoptosis in colorectal cancer cells. Journal of Agricultural and Food Chemistry. 66:9960–9967
Coppin JP, Xu Y, Chen H, Pan MH, Ho CT, Juliani R, Simon JE, Wu Q (2013) Determination of flavonoids by LC/MS and anti-inflammatory activity in Moringa oleifera. Journal of Functional Foods. 5:1892–1899
Dang Y, An Y, He J, Huang B, Zhu J, Gao M, Zhang S, Wang X, Yang B, Xie Z (2020) Berberine ameliorates cellular senescence and extends the lifespan of mice via regulating p16 and cyclin protein expression. Aging Cell. 19:1–11
Dong X, Hu X, Chen J, Hu D, Chen LF (2018) BRD4 regulates cellular senescence in gastric cancer cells via E2F/miR-106b/p21 axis. Cell Death and Disease. 9(2):1-13
Fischer KE, Gelfond JAL, Soto VY, Han C, Someya S, Richardson A, Austad SN (2015) Health effects of long-term rapamycin treatment: the impact on mouse health of enteric rapamycin treatment from four months of age throughout life. PLoS One. 10:1–18
Fitzgerald AL, Osman AA, Xie TX, Patel A, Skinner H, Sandulache V, Myers JN (2015) Reactive oxygen species and p21Waf1/Cip1 are both essential for p53-mediated senescence of head and neck cancer cells. Cell Death and Disease. 6:1–10
Gasek NS, Kuchel GA, Kirkland JL, Xu M (2021) Strategies for targeting senescent cells in human disease. Nature Aging. 1:870–879
He S, Sharpless NE (2017) Senescence in health and disease. Cell. 169(6):1000-1011
Ikawati M, Jenie RI, Utomo RY, Amalina ND, Ilmawati GPN, Kawaichi M, Meiyanto E (2020) Genistein enhances cytotoxic and antimigratory activities of doxorubicin on 4T1 breast cancer cells through cell cycle arrest and ROS generation. Journal of Applied Pharmaceutical Science. 10(10):95–104
Jenie RI, Amalina ND, Ilmawati GPN, Utomo RY, Ikawati M, Khumaira A, Kato JY, Meiyanto E (2019) Cell cycle modulation of CHO-K1 cells under genistein treatment correlates with cells senescence, apoptosis and ROS level but in a dose-dependent manner. Advanced Pharmaceutical. 9(3):453
Kornienko JS, Smirnova IS, Pugovkina NA, Ivanova JS, Shilina MA, Grinchuk TM, Shatrova, AN, Aksenov ND, Zenin VV, Nikolsky NN, Lyublinskaya OG (2019) High doses of synthetic antioxidants induce premature senescence in cultivated mesenchymal stem cells. Scientific Reports. 9(1): 1–13
Koyano T, Namba M, Kobayashi T, Nakakuni K, Nakano D, Fukushima M, Nishiyama A, Matsuyama M (2019) The p21 dependent G2 arrest of the cell cycle in epithelial tubular cells links to the early stage of renal fibrosis. Scientific Reports. 9(1):1–11
Leibiger C, Kosyakova N, Mkrtchyan H, Glei M, Trifonov V, Liehr T (2013) First molecular cytogenetic high resolution characterization of the NIH 3T3 cell line by murine multicolor banding. Journal of Histochemistry and Cytochemistry. 61(4):306–312
Leone A, Spada A, Battezzati A, Schiraldi A, Aristil J, Bertoli S (2015) Cultivation, genetic, ethnopharmacology, phytochemistry and pharmacology of Moringa oleifera leaves: an overview. International Journal of Molecular Sciences. 16(6):12791–12835
Luqman S, Srivastava S, Kumar R, Maurya AK, Chanda D (2012) Experimental assessment of Moringa oleifera leaf and fruit for its antistress, antioxidant, and scavenging potential using in vitro and in vivo assays. Evidence-based Complement and Alternative Medicine. 1:1-12
Mijit M, Caracciolo V, Melillo A, Amicarelli F, Giordano A (2020) Role of p53 in the regulation of cellular senescence. Biomolecules. 10(3):1–16
Nouman W, Anwar F, Gull T, Newton A, Rosa E, Domínguez-Perles R (2016) Profiling of polyphenolics, nutrients and antioxidant potential of germplasm’s leaves from seven cultivars of Moringa oleifera lam. Industrial Crops and Products. 83:166–176
Nurrachma MY, Maran GG, Putri NB, Esti YF, Hermawan A, Meiyanto E, Jenie RI (2020) Fingerroot (Boesenbergia pandurata) extract inhibits proliferation and migration of 4T1 metastatic breast cancer cells. Indonesian Journal of Cancer Chemoprevention. 11(3):103
Odusote JK, Owalude DO, Olusegun SJ, Yahya RA (2016) Inhibition efficiency of Moringa oleifera leaf extract on the corrosion of reinforced steel bar in HCl solution. West Indian Journal of Engineering. 38(2):64
Oladeji SO, Adelowo FE, Odelade KA (2016) Mass spectroscopic and phytochemical screening of phenolic compounds in the leaf extract of Senna alata (L.) Roxb. (Fabales: Fabaceae). Brazilian Journal of Biological Sciences. 3(5):209-219
Pantsar T, Poso A (2018) Binding affinity via docking: fact and fiction. Molecules. 23(8):1899
Pedersen JK, Engholm G, Skytthe A, Christensen, K (2016) Cancer and aging: epidemiology and methodological challenges. Acta Oncologica. 55(1):7–12
Pulakat L, Chen HH (2020). Pro-Senescence and anti-senescence mechanisms of cardiovascular aging: cardiac MicroRNA regulation of longevity drug-induced autophagy. Frontiers in Oncology. 11:1–9
Qi L, Zhou Y, Li W, Zheng M, Zhong R, Jin X, Lin Y (2019) Effect of moringa oleifera stem extract on hydrogen peroxide-induced opacity of cultured mouse lens. BMC Complementary and Alternative Medicine. 19(1):1-9
Romanov VS, Pospelov VA, Pospelova TV (2012) Cyclin-dependent kinase inhibitor p21Waf1: contemporary view on its role in senescence and oncogenesis. Biochemistry. 77(6):575–584
Salmon AB (2015) About-face on the metabolic side effects of rapamycin. Oncotarget. 6(5):2585–2586
Schosserer M, Grillari J, Breitenbach M (2017) The dual role of cellular senescence in developing tumors and their response to cancer therapy. Frontiers in Oncology. 7:278
Shtutman M, Chang BD, Schools GP, Broude EV (2017) Cellular model of p21-induced senescence. Methods in Molecular Biology. 1534:31–39
Szklarczyk D, Gable AL, Lyon D, Junge A, Wyder S, Huerta-Cepas J, Simonovic M, Doncheva NT, Morris JH, Bork P, Jensen LJ, Von Mering C (2019) STRING v11: protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets. Nucleid Acid Research. 47(D1): D607–D613
Vrancheva R, Ivanov I, Dincheva I, Badjakov I, Pavlov A (2021) Triterpenoids and other non-polar compounds in leaves of wild and cultivated Vaccinium species. Plants. 10(1):1–16
Wyld L, Bellantuono I, Tchkonia T, Morgan J, Turner O, Foss F, George J, Danson S, Kirkland JL (2020) Senescence and cancer: A review of clinical implications of senescence and senotherapies. Cancers (Basel). 12(8):1–20
Yan C, Xu Z, Huang W (2021) Cellular senescence affects cardiac regeneration and repair in ischemic heart disease. Aging and Disease. 12(2):552–569
Yong-Bing X, Gui-Lin C, Ming-Quan G (2019) Antioxidant and anti-inflammatory activities of the crude extracts of Moringa oleifera from kenya and their correlations with flavonoids. Antioxidants. 8(8):296
Zullaikah S, Naulina RY, Meinawati P, Fauziyah K, Rachimoellah M, Rachmaniah O, Nurkhamidah S, Suari NMIP, Prasetyo EN (2019) Enhanced extraction of phenolic compounds from Moringa oleifera leaves using subcritical water ethanol mixture. IOP Conference Series: Materials Science and Engineering. 543(1):1-7
Published
2023-01-31
How to Cite
Sri Andriyani, Rujitoningtyas, K., Wigati, H. U., & Amalina, N. D. (2023). The Potential of Moringa Leaf Extract to Prevent Aging Targeted Cellular Senescence. International Journal of Cell and Biomedical Science, 1(3), 76-85. https://doi.org/10.59278/cbs.v1i3.21
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