Antioxidant and cytoprotective effects of morin against hydrogen peroxide-induced oxidative stress are associated with the induction of Nrf-2‑mediated HO-1 expression in V79-4 Chinese hamster lung fibroblasts

Lee,Moon  Hee; Cha,Hee‑Jae; Choi,Eun  Ok; Han,Min  Ho; Kim,Sung  Ok; Kim,Gi‑Young; Hong,Su  Hyun; Park,Cheol; Moon,Sung‑Kwon; Jeong,Soon‑Jeong; Jeong,Moon‑Jin; Kim,Wun‑Jae; Choi,Yung  Hyun

doi:10.3892/ijmm.2017.2871

Antioxidant and cytoprotective effects of morin against hydrogen peroxide-induced oxidative stress are associated with the induction of Nrf-2‑mediated HO-1 expression in V79-4 Chinese hamster lung fibroblasts

Authors:
- Moon Hee Lee
- Hee‑Jae Cha
- Eun Ok Choi
- Min Ho Han
- Sung Ok Kim
- Gi‑Young Kim
- Su Hyun Hong
- Cheol Park
- Sung‑Kwon Moon
- Soon‑Jeong Jeong
- Moon‑Jin Jeong
- Wun‑Jae Kim
- Yung Hyun Choi
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Affiliations: Anti‑Aging Research Center, Dongeui University, Busan 614‑714, Republic of Korea, Department of Parasitology and Genetics, Kosin University, College of Medicine, Busan 602‑702, Republic of Korea, Natural Products Research Team, Marine Biodiversity Institute of Korea, Seocheon 325‑902, Republic of Korea, Department of Food Science and Biotechnology, College of Engineering, Kyungsung University, Busan 608‑736, Republic of Korea, Laboratory of Immunobiology, Department of Marine Life Sciences, Jeju National University, Jeju 690‑756, Republic of Korea, Department of Biochemistry, Dongeui University College of Korean Medicine, Busan 614‑052, Republic of Korea, Department of Molecular Biology, College of Natural Sciences and Human Ecology, Dongeui University, Busan 614‑714, Republic of Korea, School of Food Science and Technology, Chung‑Ang University, Anseong 456‑756, Republic of Korea, Department of Dental Hygiene, College of Health Sciences, Youngsan University, Yangsan 626‑790, Republic of Korea, Department of Oral Histology and Developmental Biology, School of Dentistry, Chosun University, Gwangju 501‑759, Republic of Korea, Department of Urology, Chungbuk National University, College of Medicine and Institute for Tumor Research, Cheongju 362‑763, Republic of Korea
Published online on: January 27, 2017 https://doi.org/10.3892/ijmm.2017.2871
Pages: 672-680

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Abstract

Natural phytochemicals of plant origin, including flavonoids, have been found to be potent antioxidants providing beneficial effects against oxidative stress-related diseases. The present study was carried out to investigate the antioxidant properties of morin, a flavonoid originally isolated from the flowering plants of the Moraceae family. Superoxide dismutase (SOD)‑like activity and 2,2'‑azino‑bis‑(3‑ethylbenzothiazoline‑6‑sulfonic acid) (ABTS•+) radical scavenging activity were determined. We also investigated the cytoprotective effects of morin against hydrogen peroxide (H2O2)‑induced DNA damage and apoptosis in V79‑4 Chinese hamster lung fibroblasts. Our results demonstrated that morin had strong scavenging effects against ABTS•+ radicals with enhanced SOD activity, which varied in a dose-dependent manner. Morin was found to reduce H2O2‑induced intracellular reactive oxygen species generation and nuclear DNA damage, and it recovered cell viability damaged by H2O2 via inhibition of mitochondrial dysfunction‑mediated apoptosis. Notably, the treatment of V79‑4 cells with morin markedly enhanced the expression of heme oxygenase‑1 (HO‑1) but not quinone oxidoreductase-1, which was associated with the increased expression and phosphorylation of nuclear factor-erythroid 2-related factor 2 (Nrf2) and the downregulation of Kelch‑like ECH‑associated protein 1 expression. Based on our findings, we conclude that morin effectively ameliorated oxidative stress‑induced DNA damage through intrinsic free radical scavenging activity and activation of the Nrf2/HO-1 pathway.

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1	Dröge W: Free radicals in the physiological control of cell function. Physiol Rev. 82:47–95. 2002. View Article : Google Scholar : PubMed/NCBI
2	Lyakhovich A and Graifer D: Mitochondria-mediated oxidative stress: Old Target for New drugs. Curr Med Chem. 22:3040–3053. 2015. View Article : Google Scholar : PubMed/NCBI
3	Ermakov AV, Konkova MS, Kostyuk SV, Izevskaya VL, Baranova A and Veiko NN: Oxidized extracellular DNA as a stress signal in human cells. Oxid Med Cell Longev. 2013:6497472013. View Article : Google Scholar : PubMed/NCBI
4	Yang HY and Lee TH: Antioxidant enzymes as redox-based biomarkers: A brief review. BMB Rep. 48:200–208. 2015. View Article : Google Scholar : PubMed/NCBI
5	Alfadda AA and Sallam RM: Reactive oxygen species in health and disease. J Biomed Biotechnol. 2012:9364862012. View Article : Google Scholar : PubMed/NCBI
6	Cha MY, Kim DK and Mook-Jung I: The role of mitochondrial DNA mutation on neurodegenerative diseases. Exp Mol Med. 47:e1502015. View Article : Google Scholar : PubMed/NCBI
7	Ginter E, Simko V and Panakova V: Antioxidants in health and disease. Bratisl Lek Listy. 115:603–606. 2014.
8	Cirillo G, Curcio M, Vittorio O, Iemma F, Restuccia D, Spizzirri UG, Puoci F and Picci N: Polyphenol conjugates and human health: A perspective review. Crit Rev Food Sci Nutr. 56:326–337. 2016. View Article : Google Scholar
9	Landete JM: Dietary intake of natural antioxidants: Vitamins and polyphenols. Crit Rev Food Sci Nutr. 53:706–721. 2013. View Article : Google Scholar : PubMed/NCBI
10	Kehrer JP and Klotz LO: Free radicals and related reactive species as mediators of tissue injury and disease: Implications for Health. Crit Rev Toxicol. 45:765–798. 2015. View Article : Google Scholar : PubMed/NCBI
11	Kancheva VD and Kasaikina OT: Bio-antioxidants - a chemical base of their antioxidant activity and beneficial effect on human health. Curr Med Chem. 20:4784–4805. 2013. View Article : Google Scholar : PubMed/NCBI
12	Bondonno CP, Croft KD, Ward N, Considine MJ and Hodgson JM: Dietary flavonoids and nitrate: Effects on nitric oxide and vascular function. Nutr Rev. 73:216–235. 2015. View Article : Google Scholar : PubMed/NCBI
13	Stockert JC, Colman OD and Cañete M: Fluorescence reaction of leukocyte granules by morin. Acta Histochem Suppl. 31:243–252. 1985.PubMed/NCBI
14	Srinivas NR: Recent trends in preclinical drug-drug interaction studies of flavonoids - Review of case studies, issues and perspectives. Phytother Res. 29:1679–1691. 2015. View Article : Google Scholar : PubMed/NCBI
15	Caselli A, Cirri P, Santi A and Paoli P: Morin: A Promising natural drug. Curr Med Chem. 23:774–791. 2016. View Article : Google Scholar
16	Kim JM, Lee EK, Park G, Kim MK, Yokozawa T, Yu BP and Chung HY: Morin modulates the oxidative stress-induced NF-kappaB pathway through its anti-oxidant activity. Free Radic Res. 44:454–461. 2010. View Article : Google Scholar : PubMed/NCBI
17	Komirishetty P, Areti A, Sistla R and Kumar A: Morin mitigates chronic constriction injury (CCI)-induced peripheral neuropathy by inhibiting oxidative stress induced PARP over-activation and neuroinflammation. Neurochem Res. 41:2029–2042. 2016. View Article : Google Scholar : PubMed/NCBI
18	Ola MS, Aleisa AM, Al-Rejaie SS, Abuohashish HM, Parmar MY, Alhomida AS and Ahmed MM: Flavonoid, morin inhibits oxidative stress, inflammation and enhances neurotrophic support in the brain of streptozotocin-induced diabetic rats. Neurol Sci. 35:1003–1008. 2014. View Article : Google Scholar : PubMed/NCBI
19	MadanKumar P, NaveenKumar P, Manikandan S, Devaraj H and NiranjaliDevaraj S: Morin ameliorates chemically induced liver fibrosis in vivo and inhibits stellate cell proliferation in vitro by suppressing Wnt/β-catenin signaling. Toxicol Appl Pharmacol. 277:210–220. 2014. View Article : Google Scholar : PubMed/NCBI
20	Ganguli A, Das A, Nag D, Bhattacharya S and Chakrabarti G: Potential role of autophagy in smokeless tobacco extract-induced cytotoxicity and in morin-induced protection in oral epithelial cells. Food Chem Toxicol. 90:160–170. 2016. View Article : Google Scholar : PubMed/NCBI
21	Paoli P, Cirri P, Caselli A, Ranaldi F, Bruschi G, Santi A and Camici G: The insulin-mimetic effect of Morin: A promising molecule in diabetes treatment. Biochim Biophys Acta. 1830:3102–3111. 2013. View Article : Google Scholar : PubMed/NCBI
22	Sendrayaperumal V, Iyyam Pillai S and Subramanian S: Design, synthesis and characterization of zinc-morin, a metal flavonol complex and evaluation of its antidiabetic potential in HFD-STZ induced type 2 diabetes in rats. Chem Biol Interact. 219:9–17. 2014. View Article : Google Scholar : PubMed/NCBI
23	Qureshi AA, Guan XQ, Reis JC, Papasian CJ, Jabre S, Morrison DC and Qureshi N: Inhibition of nitric oxide and inflammatory cytokines in LPS-stimulated murine macrophages by resveratrol, a potent proteasome inhibitor. Lipids Health Dis. 11:762012. View Article : Google Scholar : PubMed/NCBI
24	Dhanasekar C, Kalaiselvan S and Rasool M: Morin, a bioflavonoid suppresses monosodium urate crystal-induced inflammatory immune response in RAW 264.7 macrophages through the inhibition of inflammatory mediators, intracellular ROS levels and NF-κB activation. PLoS One. 10:e01450932015. View Article : Google Scholar
25	Dilshara MG, Jayasooriya RG, Lee S, Choi YH and Kim GY: Morin downregulates nitric oxide and prostaglandin E₂ production in LPS-stimulated BV2 microglial cells by suppressing NF-κB activity and activating HO-1 induction. Environ Toxicol Pharmacol. 44:62–68. 2016. View Article : Google Scholar : PubMed/NCBI
26	Gupta SC, Phromnoi K and Aggarwal BB: Morin inhibits STAT3 tyrosine 705 phosphorylation in tumor cells through activation of protein tyrosine phosphatase SHP1. Biochem Pharmacol. 85:898–912. 2013. View Article : Google Scholar : PubMed/NCBI
27	Manna SK, Aggarwal RS, Sethi G, Aggarwal BB and Ramesh GT: Morin (3,5,7,2′,4′-pentahydroxyflavone) abolishes nuclear factor-kappaB activation induced by various carcinogens and inflammatory stimuli, leading to suppression of nuclear factor-kappaB-regulated gene expression and up-regulation of apoptosis. Clin Cancer Res. 13:2290–2297. 2007. View Article : Google Scholar : PubMed/NCBI
28	Park C, Lee WS, Go SI, Nagappan A, Han MH, Hong SH, Kim GS, Kim GY, Kwon TK, Ryu CH, et al: Morin, a flavonoid from Moraceae, induces apoptosis by induction of BAD protein in human leukemic cells. Int J Mol Sci. 16:645–659. 2014. View Article : Google Scholar
29	Hyun HB, Lee WS, Go SI, Nagappan A, Park C, Han MH, Hong SH, Kim G, Kim GY, Cheong J, et al: The flavonoid morin from Moraceae induces apoptosis by modulation of Bcl-2 family members and Fas receptor in HCT 116 cells. Int J Oncol. 46:2670–2678. 2015.PubMed/NCBI
30	Park JY, Kang KA, Kim KC, Cha JW, Kim EH and Hyun JW: Morin induces heme oxygenase-1 via ERK-Nrf2 signaling pathway. J Cancer Prev. 18:249–256. 2013. View Article : Google Scholar
31	Rizvi F, Mathur A and Kakkar P: Morin mitigates acetaminophen-induced liver injury by potentiating Nrf2 regulated survival mechanism through molecular intervention in PHLPP2-Akt-Gsk3β axis. Apoptosis. 20:1296–1306. 2015. View Article : Google Scholar : PubMed/NCBI
32	Mathur A, Rizvi F and Kakkar P: PHLPP2 down regulation influences nuclear Nrf2 stability via Akt-1/Gsk3β/Fyn kinase axis in acetaminophen induced oxidative renal toxicity: Protection accorded by morin. Food Chem Toxicol. 89:19–31. 2016. View Article : Google Scholar : PubMed/NCBI
33	Lee YJ, Koh EK, Kim JE, Go J, Song SH, Seong JE, Son HJ, Kang BC and Hwang DY: Beneficial effects of ethanol extracts of Red Liriope platyphylla on vascular dysfunction in the aorta of spontaneously hypertensive rats. Lab Anim Res. 31:13–23. 2015. View Article : Google Scholar : PubMed/NCBI
34	Sreejayan N and Rao MN: Nitric oxide scavenging by curcuminoids. J Pharm Pharmacol. 49:105–107. 1997. View Article : Google Scholar : PubMed/NCBI
35	Kim SH, Kang SH and Kang BS: Therapeutic effects of dihydroartemisinin and transferrin against glioblastoma. Nutr Res Pract. 10:393–397. 2016. View Article : Google Scholar : PubMed/NCBI
36	Eom SA, Kim DW, Shin MJ, Ahn EH, Chung SY, Sohn EJ, Jo HS, Jeon SJ, Kim DS, Kwon HY, et al: Protective effects of PEP-1-Catalase on stress-induced cellular toxicity and MPTP-induced Parkinson's disease. BMB Rep. 48:395–400. 2015. View Article : Google Scholar :
37	Chun SK, Go K, Yang MJ, Zendejas I, Behrns KE and Kim JS: Autophagy in ischemic livers: A critical role of Sirtuin 1/Mitofusin 2 axis in autophagy induction. Toxicol Res. 32:35–46. 2016. View Article : Google Scholar : PubMed/NCBI
38	Alaoui-Jamali MA, Bismar TA, Gupta A, Szarek WA, Su J, Song W, Xu Y, Xu B, Liu G, Vlahakis JZ, et al: A novel experimental heme oxygenase-1-targeted therapy for hormone-refractory prostate cancer. Cancer Res. 69:8017–8024. 2009. View Article : Google Scholar : PubMed/NCBI
39	Hensley P, Mishra M and Kyprianou N: Targeting caspases in cancer therapeutics. Biol Chem. 394:831–843. 2013. View Article : Google Scholar : PubMed/NCBI
40	MacKenzie SH and Clark AC: Targeting cell death in tumors by activating caspases. Curr Cancer Drug Targets. 8:98–109. 2008. View Article : Google Scholar : PubMed/NCBI
41	Lazebnik YA, Kaufmann SH, Desnoyers S, Poirier GG and Earnshaw WC: Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE. Nature. 371:346–347. 1994. View Article : Google Scholar : PubMed/NCBI
42	Azqueta A, Slyskova J, Langie SA, O'Neill Gaivão I and Collins A: Comet assay to measure DNA repair: Approach and applications. Front Genet. 5:2882014. View Article : Google Scholar : PubMed/NCBI
43	Rogakou EP, Pilch DR, Orr AH, Ivanova VS and Bonner WM: DNA double-stranded breaks induce histone H2AX phosphory-lation on serine 139. J Biol Chem. 273:5858–5868. 1998. View Article : Google Scholar : PubMed/NCBI
44	Dinkova-Kostova AT, Holtzclaw WD, Cole RN, Itoh K, Wakabayashi N, Katoh Y, Yamamoto M and Talalay P: Direct evidence that sulfhydryl groups of Keap1 are the sensors regulating induction of phase 2 enzymes that protect against carcinogens and oxidants. Proc Natl Acad Sci USA. 99:11908–11913. 2002. View Article : Google Scholar : PubMed/NCBI
45	Gan L and Johnson JA: Oxidative damage and the Nrf2-ARE pathway in neurodegenerative diseases. Biochim Biophys Acta. 1842:1208–1218. 2014. View Article : Google Scholar : PubMed/NCBI
46	Wang Y, Yang J and Yi J: Redox sensing by proteins: Oxidative modifications on cysteines and the consequent events. Antioxid Redox Signal. 16:649–657. 2012. View Article : Google Scholar
47	Clerkin JS, Naughton R, Quiney C and Cotter TG: Mechanisms of ROS modulated cell survival during carcinogenesis. Cancer Lett. 266:30–36. 2008. View Article : Google Scholar : PubMed/NCBI
48	Jaramillo MC and Zhang DD: The emerging role of the Nrf2-Keap1 signaling pathway in cancer. Genes Dev. 27:2179–2191. 2013. View Article : Google Scholar : PubMed/NCBI
49	Kaspar JW and Jaiswal AK: Tyrosine phosphorylation controls nuclear export of Fyn, allowing Nrf2 activation of cytoprotective gene expression. FASEB J. 25:1076–1087. 2011. View Article : Google Scholar :
50	Jain AK and Jaiswal AK: Phosphorylation of tyrosine 568 controls nuclear export of Nrf2. J Biol Chem. 281:12132–12142. 2006. View Article : Google Scholar : PubMed/NCBI