Atrial natriuretic peptide protects vertebral endplate chondrocytes against H<sub>2</sub>O<sub>2</sub>‑induced apoptosis and oxidative stress through activation of the Nrf2/HO‑1 signaling pathway

He,Feiping; Gai,Jingying; Wang,Jun; Tang,Lei; Liu,Yifeng; Feng,Qingchun

doi:10.3892/mmr.2021.12394

Atrial natriuretic peptide protects vertebral endplate chondrocytes against H₂O₂‑induced apoptosis and oxidative stress through activation of the Nrf2/HO‑1 signaling pathway

Authors:
- Feiping He
- Jingying Gai
- Jun Wang
- Lei Tang
- Yifeng Liu
- Qingchun Feng
View Affiliations

Affiliations: Department of Spinal Surgery, The Second People's Hospital of Hunan Province, Brain Hospital of Hunan Province, Changsha, Hunan 410007, P.R. China, Department of Cardiovascular Medicine, The Second People's Hospital of Hunan Province, Brain Hospital of Hunan Province, Changsha, Hunan 410007, P.R. China
Published online on: September 1, 2021 https://doi.org/10.3892/mmr.2021.12394
Article Number: 754
Copyright: © He et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The present study aimed to investigate the effect of atrial natriuretic peptide (ANP) on cell apoptosis and oxidative stress in H₂O₂‑induced vertebral endplate chondrocytes (EPCs), and to assess the associated mechanisms involved. Cell viability and apoptosis were evaluated using the Cell Counting Kit‑8 method and TUNEL assay, respectively. In addition, the scavenging capability was detected using various enzymatic assays, and the quantity of nitric oxide (NO) and malondialdehyde (MDA), and activity of superoxide dismutase (SOD) were assessed. The expression levels of apoptosis‑related proteins, activation of the nuclear factor erythroid 2‑related factor 2 (Nrf2)/heme oxygenase‑1 (HO‑1) signaling pathway induced by H₂O₂ and the effect of treatment with ANP on vertebral EPCs were detected by western blotting. The results revealed that ANP protected EPCs from H₂O₂‑induced cell damage. H₂O₂‑induced intracellular MDA was decreased by ANP, and the levels of SOD and NO were increased in the presence of ANP. ANP also inhibited the H₂O₂‑induced alterations in the expression levels of cleaved‑caspase‑3, Bax and Bcl‑2. Finally, ANP blocked H2O2‑induced oxidative stress through activating the Nrf2/HO‑1 signaling pathway. These findings suggested that ANP may effectively protect EPCs through inhibition of H₂O₂‑induced oxidant injury and cell death by activating the Nrf2/HO‑1 signaling pathway.

View Figures

View References

1	Hu RM, Levin ER, Pedram A and Frank HJL: Atrial natriuretic peptide inhibits the production and secretion of endothelin from cultured endothelial cells: Mediation through the C receptor. J Biol Chem. 267:17384–9. 1992. View Article : Google Scholar : PubMed/NCBI
2	Smith S, Anderson S, Ballermann BJ and Brenner BM: Role of atrial natriuretic peptide in adaptation of sodium excretion with reduced renal mass. J Clin Invest. 77:1395–1398. 1986. View Article : Google Scholar : PubMed/NCBI
3	Baxter JD, Lewicki JA and Gardner DG: Atrial Natriuretic Peptide. Nat Biotechnol. 6:529–546. 1988. View Article : Google Scholar
4	Abraham WT, Hensen J, Kim JK, Dürr J and Schrier RW: Atrial natriuretic peptide and urinary cyclic guanosine monophosphate in patients with chronic heart failure. J Am Soc Nephrol. 2:1697–1703. 1992. View Article : Google Scholar : PubMed/NCBI
5	Rahman SN, Kim GE, Mathew AS, Goldberg CA, Allgren R, Schrier RW and Conger JD: Effects of atrial natriuretic peptide in clinical acute renal failure. Kidney Int. 45:1731–1738. 1994. View Article : Google Scholar : PubMed/NCBI
6	Bilzer M, Jaeschke H, Vollmar AM, Paumgartner G and Gerbes AL: Prevention of Kupffer cell-induced oxidant injury in rat liver by atrial natriuretic peptide. Am J Physiol. 276:G1137–G1144. 1999.PubMed/NCBI
7	Bilzer M, Jaeschke H, Vollmar AM, Paumgartner G and Gerbes AL: Reduction of Kupffer cell-induced oxidant injury of the rat liver by atrial natriuretic peptide and cyclo-GMP receptor proteins. J Hepatol. 28:471998. View Article : Google Scholar
8	Fahn S and Cohen G: The oxidant stress hypothesis in Parkinson's disease: Evidence supporting it. Ann Neurol. 32:804–812. 1992. View Article : Google Scholar : PubMed/NCBI
9	Jie B, Fan Y, Li D and Yi Z: Ghrelin protects human lens epithelial cells against oxidative stress-induced damage. Oxid Med Cell Longev. 2017:19104502017.PubMed/NCBI
10	Martinou JC and Youle RJ: Mitochondria in apoptosis: Bcl-2 family members and mitochondrial dynamics. Dev Cell. 21:92–101. 2011. View Article : Google Scholar : PubMed/NCBI
11	Kluck RM, Bossy-Wetzel E, Green DR and Newmeyer DD: The release of cytochrome c from mitochondria: A primary site for Bcl-2 regulation of apoptosis. Science. 275:1132–1136. 1997. View Article : Google Scholar : PubMed/NCBI
12	Shi L, Chen J, Yang J, Pan T, Zhang S and Wang Z: miR-21 protected human glioblastoma U87MG cells from chemotherapeutic drug temozolomide induced apoptosis by decreasing Bax/Bcl-2 ratio and caspase-3 activity. Brain Res. 1352:255–264. 2010. View Article : Google Scholar : PubMed/NCBI
13	Kensler TW, Wakabayashi N and Biswal S: Cell Survival responses to environmental stresses via the Keap1-Nrf2-ARE Pathway. Annu Rev Pharmacol Toxicol. 47:89–116. 2007. View Article : Google Scholar : PubMed/NCBI
14	Mukaigasa K, Nguyen LTP, Li L, Nakajima H, Yamamoto M and Kobayashi M: Genetic evidence of an evolutionarily conserved role for Nrf2 in the protection against oxidative stress. Mol Cell Biol. 32:4455–4461. 2012. View Article : Google Scholar : PubMed/NCBI
15	Strom J: A critical role of Nrf2 in protecting cardiomyocytes against oxidative stress and ischemic injury. PhD dissertation, The University of Arizona, . 2014.
16	Buendia I, Michalska P, Navarro E, Gameiro I and León R: Nrf2-ARE pathway: An emerging target against oxidative stress and neuroinflammation in neurodegenerative diseases. Pharmacol Ther. 157:84–104. 2016. View Article : Google Scholar : PubMed/NCBI
17	Yagishita Y, Fukutomi T, Sugawara A, Kawamura H, Takahashi T, Pi J, Uruno A and Yamamoto M: Nrf2 protects pancreatic beta-cells from oxidative and nitrosative stress in diabetic model mice. Diabetes. 63:605–618. 2014. View Article : Google Scholar : PubMed/NCBI
18	Loboda A, Damulewicz M, Pyza E, Jozkowicz A and Dulak J: Role of Nrf2/HO-1 system in development, oxidative stress response and diseases: An evolutionarily conserved mechanism. Cell Mol Life Sci. 73:3221–3247. 2016. View Article : Google Scholar : PubMed/NCBI
19	Jian Z, Li K, Liu L, Zhang Y, Zhou Z, Li C and Gao T: Heme Oxygenase-1 protects human melanocytes from H₂O₂-induced oxidative stress via the Nrf2-ARE pathway. J Invest Dermatol. 131:1420–1427. 2011. View Article : Google Scholar : PubMed/NCBI
20	Sahin K, Tuzcu M, Sahin N, Ali S and Kucuk O: Nrf2/HO-1 signaling pathway may be the prime target for chemoprevention of cisplatin-induced nephrotoxicity by lycopene. Food Chem Toxicol. 48:2670–2674. 2010. View Article : Google Scholar : PubMed/NCBI
21	Han Y, Li X, Yan M, Yang M, Wang S, Pan J, Li L and Tan J: Oxidative damage induces apoptosis and promotes calcification in disc cartilage endplate cell through ROS/MAPK/NF-κB pathway: Implications for disc degeneration. Biochem Biophys Res Commun. 516:1026–1032. 2019. View Article : Google Scholar : PubMed/NCBI
22	Giers MB, Munter BT, Eyster KJ, Ide GD, Newcomb AGUS, Lehrman JN, Belykh E, Byvaltsev VA, Kelly BP, Preul MC and Theodore N: Biomechanical and endplate effects on nutrient transport in the intervertebral disc. World Neurosurg. 99:395–402. 2017. View Article : Google Scholar : PubMed/NCBI
23	Ariga K, Yonenobu K, Nakase T, Hosono N, Okuda S, Meng W, Tamura Y and Yoshikawa H: Mechanical stress-induced apoptosis of endplate chondrocytes in organ-cultured mouse intervertebral discs: An ex vivo study. Spine (Phila Pa 1976). 28:1528–1533. 2003. View Article : Google Scholar : PubMed/NCBI
24	Otero M, Lago R, Lago F, Reino JJG and Gualillo O: Signalling pathway involved in nitric oxide synthase type II activation in chondrocytes: Synergistic effect of leptin with interleukin-1. Arthritis Res Ther. 7:R581–R591. 2005. View Article : Google Scholar : PubMed/NCBI
25	Zhang JX, Han YP, Bai C and Li Q: Notch1/3 and p53/p21 are a potential therapeutic target for APS-induced apoptosis in non-small cell lung carcinoma cell lines. Int J Clin Exp Med. 8:12539–12547. 2015.PubMed/NCBI
26	Zhang Z, Lin J, Tian N, Wu Y, Zhou Y, Wang C, Wang Q, Jin H, Chen T, Nisar M, et al: Melatonin protects vertebral endplate chondrocytes against apoptosis and calcification via the Sirt1-autophagy pathway. J Cell Mol Med. 23:177–193. 2019. View Article : Google Scholar : PubMed/NCBI
27	Zhou YF, Guo B, Ye MJ, Liao RF and Li SL: Protective effect of rutin against H₂O₂-induced oxidative stress and apoptosis in human lens epithelial cells. Curr Eye Res. 41:933–942. 2016. View Article : Google Scholar : PubMed/NCBI
28	Liang B, Wei W, Wang J, Zhang M, Xu R, Wu F, Xiao H and Tang L: Protective effects of Semiaquilegia adoxoides n-butanol extract against hydrogen peroxide-induced oxidative stress in human lens epithelial cells. Pharm Biol. 54:1656–1663. 2016. View Article : Google Scholar : PubMed/NCBI
29	Salakou S, Kardamakis D, Tsamandas AC, Zolota V, Apostolakis E, Tzelepi V, Papathanasopoulos P, Bonikos DS, Papapetropoulos T, Petsas T and Dougenis D: Increased Bax/Bcl-2 ratio up-regulates caspase-3 and increases apoptosis in the thymus of patients with myasthenia gravis. In Vivo. 21:123–132. 2007.PubMed/NCBI
30	Ghate NB, Hazra B, Sarkar R, Chaudhuri D and Mandal N: Alteration of Bax/Bcl-2 ratio contributes to Terminalia Belerica-induced apoptosis in human lung and breast carcinoma. In Vitro Cell Dev Biol Anim. 50:527–537. 2014. View Article : Google Scholar : PubMed/NCBI
31	Chen HM, Lai ZQ, Liao HJ, Xie JH, Xian YF, Chen YL, Ip SP, Lin ZX and Su ZR: Synergistic antitumor effect of brusatol combined with cisplatin on colorectal cancer cells. Int J Mol Med. 41:1447–1454. 2018.PubMed/NCBI
32	Hart BA: Characterization of cadmium-induced apoptosis in rat lung epithelial cells: Evidence for the participation of oxidant stress. Toxicology. 133:43–58. 1999. View Article : Google Scholar : PubMed/NCBI
33	Olayanju A, Copple IM, Bryan HK, Edge GT, Sison RL, Wong MW, Lai ZQ, Lin ZX, Dunn K, Sanderson CM, et al: Brusatol provokes a rapid and transient inhibition of Nrf2 signaling and sensitizes mammalian cells to chemical toxicity-implications for therapeutic targeting of Nrf2. Free Radic Biol Med. 78:202–212. 2015. View Article : Google Scholar : PubMed/NCBI
34	El-Beltagi HS and Mohamed HI: Reactive oxygen species, lipid peroxidation and antioxidative defense mechanism. Notulae Botanicae Horti Agrobotanici Cluj-Napoca. 41:44–57. 2013. View Article : Google Scholar
35	Zheng Y, Liu Y, Ge J, Wang X and Liu P: Resveratrol protects human lens epithelial cells against H₂O₂-induced oxidative stress by increasing catalase, SOD-1, and HO-1 expression. Mol Vis. 16:1467–1474. 2010.PubMed/NCBI
36	Velichkova M and Hasson T: Keap1 regulates the oxidation-sensitive shuttling of Nrf2 into and out of the nucleus via a Crm1-dependent nuclear export mechanism. Mol Cell Biol. 25:4501–4513. 2005. View Article : Google Scholar : PubMed/NCBI
37	Camara NO and Soares MP: Heme oxygenase-1 (HO-1), a protective gene that prevents chronic graft dysfunction. Free Radic Biol Med. 38:426–435. 2005. View Article : Google Scholar : PubMed/NCBI
38	Abraham NG and Kappas A: Heme oxygenase and the cardiovascular-renal system. Free Radic Biol Med. 39:1–25. 2005. View Article : Google Scholar : PubMed/NCBI
39	Ren D, Villeneuve NF, Jiang T, Wu T, Lau A, Toppin HA and Zhang DD: Brusatol enhances the efficacy of chemotherapy by inhibiting the Nrf2-mediated defense mechanism. Proc Natl Acad Sci USA. 108:1433–1438. 2011. View Article : Google Scholar : PubMed/NCBI
40	Tang XQ, Feng JQ, Chen J, Chen PX, Zhi JL, Cui Y, Guo RX and Yu HM: Protection of oxidative preconditioning against apoptosis induced by H₂O₂ in PC12 cells: Mechanisms via MMP, ROS, and Bcl-2. Brain Res. 1057:57–64. 2005. View Article : Google Scholar : PubMed/NCBI
41	Buttke TM and Sandstrom P: Oxidative stress as a mediator of apoptosis. Immunol Today. 15:7–10. 1994. View Article : Google Scholar : PubMed/NCBI