The cytoprotective effects of 7,8-dihydroxyflavone against oxidative stress are mediated by the upregulation of Nrf2-dependent HO-1 expression through the activation of the PI3K/Akt and ERK pathways in C2C12 myoblasts

Kang,Ji  Sook; Choi,Il-Whan; Han,Min  Ho; Kim,Gi-Young; Hong,Su  Hyun; Park,Cheol; Hwang,Hye  Jin; Kim,Cheol  Min; Kim,Byung  Woo; Choi,Yung  Hyun

doi:10.3892/ijmm.2015.2256

The cytoprotective effects of 7,8-dihydroxyflavone against oxidative stress are mediated by the upregulation of Nrf2-dependent HO-1 expression through the activation of the PI3K/Akt and ERK pathways in C2C12 myoblasts

Authors:
- Ji Sook Kang
- Il-Whan Choi
- Min Ho Han
- Gi-Young Kim
- Su Hyun Hong
- Cheol Park
- Hye Jin Hwang
- Cheol Min Kim
- Byung Woo Kim
- Yung Hyun Choi
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Affiliations: Blue-Bio Industry RIC and Anti-Aging Research Center, Dongeui University, Busan 614-714, Republic of Korea, Department of Microbiology, College of Medicine, Inje University, Busan 608-756, 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, Department of Biochemistry, Busan National University College of Medicine, Yangsan, Gyeongsangnam-do 626-870, Republic of Korea
Published online on: June 22, 2015 https://doi.org/10.3892/ijmm.2015.2256
Pages: 501-510

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Abstract

Recent studies have demonstrated that 7,8-dihydroxyflavone (7,8-DHF), a newly identified tyrosine kinase receptor B agonist, is a potent antioxidant agent. The present study was designed to confirm the cytoprotective effects of 7,8‑DHF against oxidative stress‑induced cellular damage and to further elucidate the underlying mechanisms in C2C12 myoblasts. We found that 7,8‑DHF attenuated hydrogen peroxide (H2O2)‑induced growth inhibition and exhibited scavenging activity against intracellular reactive oxygen species (ROS) that were induced by H2O2. We also observed that 7,8‑DHF significantly attenuated H2O2‑induced comet tail formation, and decreased the phosphorylation levels of the histone, H2AX, as well as the number of Annexin V‑positive cells, suggesting that 7,8‑DHF prevents H2O2‑induced DNA damage and cell apoptosis. Furthermore, 7,8‑DHF increased the levels of heme oxygenase‑1 (HO‑1), which is a potent antioxidant enzyme associated with the induction and phosphorylation of nuclear factor‑erythroid 2‑related factor 2 (Nrf2), as well as the translocation of Nrf2 from the cytosol to the nucleus. However, the protective effects of 7,8‑DHF against H2O2‑induced ROS generation and growth inhibition were significantly diminished by zinc protoporphyrin IX, an HO‑1 competitive inhibitor. Moreover, the potential of 7,8‑DHF to mediate HO‑1 induction and protect the cells against H2O2‑mediated growth inhibition was abrogated by transient transfection with Nrf2‑specific small interfering RNA (siRNA). In addition, 7,8‑DHF induced the activation of Akt, a downstream target of phosphatidylinositol 3‑kinase (PI3K), and also that of extracellular signal‑regulated kinase (ERK) and p38 mitogen‑activated protein kinase (MAPK), while specific inhibitors of PI3K and ERK, but not a p38 MAPK inhibitor, abolished the 7,8‑DHF induced HO‑1 upregulation and Nrf2 induction and phosphorylation. Collectively, these results demonstrate that 7,8‑DHF augments the cellular antioxidant defense capacity through activation of the Nrf2/HO‑1 pathway, which also involves the activation of the PI3K/Akt and ERK pathways, thereby protecting C2C12 myoblasts from H2O2-induced oxidative cytotoxicity.

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1	Kregel KC and Zhang HJ: An integrated view of oxidative stress in aging: basic mechanisms, functional effects, and pathological considerations. Am J Physiol Regul Integr Comp Physiol. 292:R18–R36. 2007. View Article : Google Scholar
2	Finkel T: Signal transduction by reactive oxygen species. J Cell Biol. 194:7–15. 2011. View Article : Google Scholar : PubMed/NCBI
3	Venugopal R and Jaiswal AK: Nrf1 and Nrf2 positively and c-Fos and Fra1 negatively regulate the human antioxidant response element-mediated expression of NAD(P)H:quinone oxidoreductase1 gene. Proc Natl Acad Sci USA. 93:14960–14965. 1996. View Article : Google Scholar : PubMed/NCBI
4	Zhang Y and Gordon GB: A strategy for cancer prevention: Stimulation of the Nrf2-ARE signaling pathway. Mol Cancer Ther. 3:885–893. 2004.PubMed/NCBI
5	Kaspar JW, Niture SK and Jaiswal AK: Nrf2:INrf2 (Keap1) signaling in oxidative stress. Free Radic Biol Med. 47:1304–1309. 2009. View Article : Google Scholar : PubMed/NCBI
6	Niture SK, Khatri R and Jaiswal AK: Regulation of Nrf2-an update. Free Radic Biol Med. 66:36–44. 2014. View Article : Google Scholar
7	Surh YJ, Kundu JK and Na HK: Nrf2 as a master redox switch in turning on the cellular signaling involved in the induction of cytoprotective genes by some chemopreventive phytochemicals. Planta Med. 74:1526–1539. 2008. View Article : Google Scholar : PubMed/NCBI
8	Kweon MH, In Park Y, Sung HC and Mukhtar H: The novel antioxidant 3-O-caffeoyl-1-methylquinic acid induces Nrf2-dependent phase II detoxifying genes and alters intracellular glutathione redox. Free Radic Biol Med. 40:1349–1361. 2006. View Article : Google Scholar : PubMed/NCBI
9	Jaiswal AK: Nrf2 signaling in coordinated activation of antioxidant gene expression. Free Radic Biol Med. 36:1199–1207. 2004. View Article : Google Scholar : PubMed/NCBI
10	Wang Y, Chun OK and Song WO: Plasma and dietary antioxidant status as cardiovascular disease risk factors: a review of human studies. Nutrients. 5:2969–3004. 2013. View Article : Google Scholar : PubMed/NCBI
11	Liu X, Chan CB, Jang SW, Pradoldej S, Huang J, He K, Phun LH, France S, Xiao G, Jia Y, et al: A synthetic 7,8-dihydroxyflavone derivative promotes neurogenesis and exhibits potent antidepressant effect. J Med Chem. 53:8274–8286. 2010. View Article : Google Scholar : PubMed/NCBI
12	Devi L and Ohno M: 7,8-Dihydroxyflavone, a small-molecule TrkB agonist, reverses memory deficits and BACE1 elevation in a mouse model of Alzheimer’s disease. Neuropsychopharmacology. 37:434–444. 2012. View Article : Google Scholar :
13	Gupta VK, You Y, Li JC, Klistorner A and Graham SL: Protective effects of 7,8-dihydroxyflavone on retinal ganglion and RGC-5 cells against excitotoxic and oxidative stress. J Mol Neurosci. 49:96–104. 2013. View Article : Google Scholar
14	Chen J, Chua KW, Chua CC, Yu H, Pei A, Chua BH, Hamdy RC, Xu X and Liu CF: Antioxidant activity of 7,8-dihydroxyflavone provides neuroprotection against glutamate-induced toxicity. Neurosci Lett. 499:181–185. 2011. View Article : Google Scholar : PubMed/NCBI
15	Zhang R, Kang KA, Piao MJ, Ko DO, Wang ZH, Chang WY, You HJ, Lee IK, Kim BJ, Kang SS and Hyun JW: Preventive effect of 7,8-dihydroxyflavone against oxidative stress induced genotoxicity. Biol Pharm Bull. 32:166–171. 2009. View Article : Google Scholar : PubMed/NCBI
16	Han XH, Cheng MN, Chen L, Fang H, Wang LJ, Li XT and Qu ZQ: 7,8-Dihydroxyflavone protects PC12 cells against 6-hydroxydopamine-induced cell death through modulating PI3K/Akt and JNK pathways. Neurosci Lett. 581:85–88. 2014. View Article : Google Scholar : PubMed/NCBI
17	Park HY, Park C, Hwang HJ, Kim BW, Kim GY, Kim CM, Kim ND and Choi YH: 7,8-Dihydroxyflavone attenuates the release of pro-inflammatory mediators and cytokines in lipopolysaccharide-stimulated BV2 microglial cells through the suppression of the NF-κB and MAPK signaling pathways. Int J Mol Med. 33:1027–1034. 2014.PubMed/NCBI
18	Park HY, Kim GY, Hyun JW, Hwang HJ, Kim ND, Kim BW and Choi YH: 7,8-Dihydroxyflavone exhibits anti-inflammatory properties by downregulating the NF-κB and MAPK signaling pathways in lipopolysaccharide-treated RAW264.7 cells. Int J Mol Med. 29:1146–1152. 2012.PubMed/NCBI
19	Ryu MJ, Kang KA, Piao MJ, Kim KC, Zheng J, Yao CW, Cha JW, Chung HS, Kim SC, Jung E, et al: 7,8-Dihydroxyflavone protects human keratinocytes against oxidative stress-induced cell damage via the ERK and PI3K/Akt-mediated Nrf2/HO-1 signaling pathways. Int J Mol Med. 33:964–970. 2014.PubMed/NCBI
20	Ryu MJ, Kang KA, Piao MJ, Kim KC, Zheng J, Yao CW, Cha JW, Hyun CL, Chung HS, Park JC, et al: Effect of 7, 8-dihydroxy-flavone on the upregulation of Nrf2-mediated heme oxygenase-1 expression in hamster lung fibroblasts. In Vitro Cell Dev Biol Anim. 50:549–554. 2014. View Article : Google Scholar : PubMed/NCBI
21	Kim KC, Lee IK, Kang KA, Cha JW, Cho SJ, Na SY, Chae S, Kim HS, Kim S and Hyun JW: 7,8-Dihydroxyflavone suppresses oxidative stress-induced base modification in DNA via induction of the repair enzyme 8-oxoguanine DNA glycosylase-1. Biomed Res Int. 2013:8637202013. View Article : Google Scholar : PubMed/NCBI
22	Kim YS, Li XF, Kang KH, Ryu B and Kim SK: Stigmasterol isolated from marine microalgae Navicula incerta induces apoptosis in human hepatoma HepG2 cells. BMB Rep. 47:433–438. 2014. View Article : Google Scholar :
23	Song JL, Choi JH, Seo JH, Kil JH and Park KY: Antioxidative effects of fermented sesame sauce against hydrogen peroxide-induced oxidative damage in LLC-PK1 porcine renal tubule cells. Nutr Res Pract. 8:138–145. 2014. View Article : Google Scholar : PubMed/NCBI
24	Jiang R, Teng Y, Huang Y, Gu J, Ma L, Li M and Zhou Y: Preeclampsia serum-induced collagen I expression and intracellular calcium levels in arterial smooth muscle cells are mediated by the PLC-γ1 pathway. Exp Mol Med. 46:e1152014. View Article : Google Scholar
25	Rogakou EP, Pilch DR, Orr AH, Ivanova VS and Bonner WM: DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139. J Biol Chem. 273:5858–5868. 1998. View Article : Google Scholar : PubMed/NCBI
26	Pischke SE, Zhou Z, Song R, Ning W, Alam J, Ryter SW and Choi AM: Phosphatidylinositol 3-kinase/Akt pathway mediates heme oxygenase-1 regulation by lipopolysaccharide. Cell Mol Biol (Noisy-le-grand). 51:461–470. 2005.
27	Paine A, Eiz-Vesper B, Blasczyk R and Immenschuh S: Signaling to heme oxygenase-1 and its anti-inflammatory therapeutic potential. Biochem Pharmacol. 80:1895–1903. 2010. View Article : Google Scholar : PubMed/NCBI
28	Yang JJ, Tao H, Huang C and Li J: Nuclear erythroid 2-related factor 2: a novel potential therapeutic target for liver fibrosis. Food Chem Toxicol. 59:421–427. 2013. View Article : Google Scholar : PubMed/NCBI
29	Woldu AS and Mai J: A novel relationship between the radical-scavenging activity of flavonoids and enthalpy of formation revealed with Hartree-Fock computations and thermochemical deduction. Redox Rep. 17:115–130. 2012. View Article : Google Scholar : PubMed/NCBI
30	Melidou M, Riganakos K and Galaris D: Protection against nuclear DNA damage offered by flavonoids in cells exposed to hydrogen peroxide: the role of iron chelation. Free Radic Biol Med. 39:1591–1600. 2005. View Article : Google Scholar : PubMed/NCBI
31	Chapple SJ, Siow RC and Mann GE: Crosstalk between Nrf2 and the proteasome: therapeutic potential of Nrf2 inducers in vascular disease and aging. Int J Biochem Cell Biol. 44:1315–1320. 2012. View Article : Google Scholar : PubMed/NCBI
32	Chen K and Keaney J: Reactive oxygen species-mediated signal transduction in the endothelium. Endothelium. 11:109–121. 2004. View Article : Google Scholar : PubMed/NCBI
33	Mo C, Wang L, Zhang J, Numazawa S, Tang H, Tang X, Han X, Li J, Yang M, Wang Z, et al: The crosstalk between Nrf2 and AMPK signal pathways is important for the anti-inflammatory effect of berberine in LPS-stimulated macrophages and endotoxin-shocked mice. Antioxid Redox Signal. 20:574–588. 2014. View Article : Google Scholar :
34	Deng C, Sun Z, Tong G, Yi W, Ma L, Zhao B, Cheng L, Zhang J, Cao F and Yi D: α-Lipoic acid reduces infarct size and preserves cardiac function in rat myocardial ischemia/reperfusion injury through activation of PI3K/Akt/Nrf2 pathway. PLoS One. 8:e583712013. View Article : Google Scholar
35	Chen HH, Chen YT, Huang YW, Tsai HJ and Kuo CC: 4-Ketopinoresinol, a novel naturally occurring ARE activator, induces the Nrf2/HO-1 axis and protects against oxidative stress-induced cell injury via activation of PI3K/AKT signaling. Free Radic Biol Med. 52:1054–1066. 2012. View Article : Google Scholar : PubMed/NCBI
36	Jeon WK, Hong HY and Kim BC: Genipin upregulates heme oxygenase-1 via PI3-kinase-JNK1/2-Nrf2 signaling pathway to enhance the anti-inflammatory capacity in RAW264.7 macrophages. Arch Biochem Biophys. 512:119–125. 2011. View Article : Google Scholar : PubMed/NCBI
37	Xu C, Yuan X, Pan Z, Shen G, Kim JH, Yu S, Khor TO, Li W, Ma J and Kong AN: Mechanism of action of isothiocyanates: The induction of ARE-regulated genes is associated with activation of ERK and JNK and the phosphorylation and nuclear translocation of Nrf2. Mol Cancer Ther. 5:1918–1926. 2006. View Article : Google Scholar : PubMed/NCBI
38	Koriyama Y, Nakayama Y, Matsugo S and Kato S: Protective effect of lipoic acid against oxidative stress is mediated by Keap1/Nrf2-dependent heme oxygenase-1 induction in the RGC-5 cell line. Brain Res. 1499:145–157. 2013. View Article : Google Scholar : PubMed/NCBI
39	Jiang G, Hu Y, Liu L, Cai J, Peng C and Li Q: Gastrodin protects against MPP(+)-induced oxidative stress by up regulates heme oxygenase-1 expression through p38 MAPK/Nrf2 pathway in human dopaminergic cells. Neurochem Int. 75:79–88. 2014. View Article : Google Scholar : PubMed/NCBI
40	Lee DS, Kim KS, Ko W, Li B, Jeong GS, Jang JH, Oh H and Kim YC: The cytoprotective effect of sulfuretin against tert-butyl hydroperoxide-induced hepatotoxicity through Nrf2/ARE and JNK/ERK MAPK-mediated heme oxygenase-1 expression. Int J Mol Sci. 15:8863–8877. 2014. View Article : Google Scholar : PubMed/NCBI
41	Jun YJ, Lee M, Shin T, Yoon N, Kim JH and Kim HR: eckol enhances heme oxygenase-1 expression through activation of Nrf2/JNK pathway in HepG2 cells. Molecules. 19:15638–15652. 2014. View Article : Google Scholar : PubMed/NCBI
42	Lee J and Kim S: Upregulation of heme oxygenase-1 expression by dehydrodiconiferyl alcohol (DHCA) through the AMPK-Nrf2 dependent pathway. Toxicol Appl Pharmacol. 281:87–100. 2014. View Article : Google Scholar : PubMed/NCBI
43	Boutros T, Chevet E and Metrakos P: Mitogen-activated protein (MAP) kinase/MAP kinase phosphatase regulation: Rroles in cell growth, death, and cancer. Pharmacol Rev. 60:261–310. 2008. View Article : Google Scholar : PubMed/NCBI