Calcium chelator BAPTA‑AM protects against iron overload‑induced chondrocyte mitochondrial dysfunction and cartilage degeneration

Jing,Xingzhi; Wang,Qiang; Du,Ting; Zhang,Weimin; Liu,Xiaoyang; Liu,Qiang; Li,Tao; Wang,Guodong; Chen,Feifei; Cui,Xingang

doi:10.3892/ijmm.2021.5029

Calcium chelator BAPTA‑AM protects against iron overload‑induced chondrocyte mitochondrial dysfunction and cartilage degeneration

Authors:
- Xingzhi Jing
- Qiang Wang
- Ting Du
- Weimin Zhang
- Xiaoyang Liu
- Qiang Liu
- Tao Li
- Guodong Wang
- Feifei Chen
- Xingang Cui
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Affiliations: Department of Spine Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong 250021, P.R. China, Department of Human Resources, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong 250021, P.R. China, Yidu Cloud (Beijing) Technology Co., Ltd., Beijing 100191, P.R. China
Published online on: August 31, 2021 https://doi.org/10.3892/ijmm.2021.5029
Article Number: 196
Copyright: © Jing et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Osteoarthritis (OA) is a common joint disease that is characterized by cartilage degradation. Iron deposition in the joints is common during the pathogenic progression of OA and recent studies have indicated that iron overload is an important contributor to OA progression. Calcium chelators have been reported to inhibit iron influx via modulating transferrin receptor protein 1 internalization, and they have been identified as a potential approach to the treatment of iron overload‑induced diseases. The aim of the present study was to investigate the effect of calcium chelators on the progression of iron overload‑induced OA. Primary chondrocytes were treated with various concentrations of ferric ammonium citrate (FAC) to mimic iron overload in vitro, followed by co‑treatment with the calcium chelator BAPTA acetoxymethyl ester (BAPTA‑AM). Subsequently, intracellular iron levels, cell viability, reactive oxygen species (ROS) levels, mitochondrial function and morphological changes, as well as MMP levels, were detected using commercial kits. It was demonstrated that FAC treatment significantly promoted chondrocyte apoptosis and the expression of MMPs, and these effects were reversed by co‑treatment with BAPTA‑AM. Moreover, BAPTA‑AM suppressed iron influx into chondrocytes and inhibited iron overload‑induced ROS production and mitochondrial dysfunction. These results indicated that calcium chelators may be of value in the treatment of iron metabolism‑related diseases and iron overload‑induced OA progression.

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1	Jing X, Du T, Li T, Yang X, Wang G, Liu X, Jiang Z and Cui X: The detrimental effect of iron on OA chondrocytes: Importance of pro-inflammatory cytokines induced iron influx and oxidative stress. J Cell Mol Med. 25:5671–5680. 2021. View Article : Google Scholar : PubMed/NCBI
2	Jing X, Lin J, Du T, Jiang Z, Li T, Wang G, Liu X, Cui X and Sun K: Iron overload is associated with accelerated progression of osteoarthritis: The role of DMT1 mediated iron homeostasis. Front Cell Dev Biol. 8:5945092020. View Article : Google Scholar
3	Jeney V: Clinical impact and cellular mechanisms of iron overload-associated bone loss. Front Pharmacol. 8:772017. View Article : Google Scholar : PubMed/NCBI
4	Sheth S: Iron chelation: An update. Curr Opin Hematol. 21:179–185. 2014. View Article : Google Scholar : PubMed/NCBI
5	Gozzelino R and Arosio P: Iron homeostasis in health and disease. Int J Mol Sci. 17:1302016. View Article : Google Scholar :
6	Leipuviene R and Theil E: The family of iron responsive RNA structures regulated by changes in cellular iron and oxygen. Cell Mol Life Sci. 64:2945–2955. 2007. View Article : Google Scholar : PubMed/NCBI
7	Xie Y, Hou W, Song X, Yu Y, Huang J, Sun X, Kang R and Tang D: Ferroptosis: Process and function. Cell Death Differe. 23:369–379. 2016. View Article : Google Scholar
8	Siddique A and Kowdley KV: Review article: The iron overload syndromes. Aliment Pharmacol Ther. 35:876–893. 2012. View Article : Google Scholar : PubMed/NCBI
9	Ogilvie-Harris DJ and Fornaiser VL: Synovial iron deposition in osteoarthritis and rheumatoid arthritis. J Rheumatol. 7:30–36. 1980.PubMed/NCBI
10	Roosendaal G, Tekoppele JM, Vianen ME, van den Berg HM, Lafeber FP and Bijlsma JW: Articular cartilage is more susceptible to blood induced damage at young than at old age. J Rheumatol. 27:1740–1744. 2000.PubMed/NCBI
11	Naughton DP: Iron(III)-mediated intra-articular crystal deposition in arthritis: A therapeutic role for iron chelators. Med Hypotheses. 57:120–122. 2001. View Article : Google Scholar : PubMed/NCBI
12	Nakamura T, Naguro I and Ichijo H: Iron homeostasis and iron-regulated ROS in cell death, senescence and human diseases. Biochim Biophys Acta. 1863:1398–1409. 2019. View Article : Google Scholar
13	Kennish L, Attur M, Oh C, Krasnokutsky S, Samuels J, Greenberg JD, Huang X and Abramson SB: Age-dependent ferritin elevations and HFE C282Y mutation as risk factors for symptomatic knee osteoarthritis in males: A longitudinal cohort study. BMC Musculoskelet Disord. 15:82014. View Article : Google Scholar : PubMed/NCBI
14	Suantawee T, Tantavisut S, Adisakwattana S, Tanavalee A, Yuktanandana P, Anomasiri W, Deepaisarnsakul B and Honsawek S: Oxidative stress, vitamin e, and antioxidant capacity in knee osteoarthritis. J Clin Diagn Res. 7:1855–1859. 2013.PubMed/NCBI
15	Blanco FJ, Rego I and Ruiz-Romero C: The role of mitochondria in osteoarthritis. Nat Rev Rheumatol. 7:161–169. 2011. View Article : Google Scholar : PubMed/NCBI
16	Kühn LC: Iron regulatory proteins and their role in controlling iron metabolism. Metallomics. 7:232–243. 2015. View Article : Google Scholar
17	Kumfu S, Chattipakorn SC, Fucharoen S and Chattipakorn N: Dual T-type and L-type calcium channel blocker exerts beneficial effects in attenuating cardiovascular dysfunction in iron-overloaded thalassaemic mice. Exp Physiol. 101:521–539. 2016. View Article : Google Scholar : PubMed/NCBI
18	Sripetchwandee J, KenKnight SB, Sanit J, Chattipakorn S and Chattipakorn N: Blockade of mitochondrial calcium uniporter prevents cardiac mitochondrial dysfunction caused by iron overload. Acta Physiol (Oxf). 210:330–341. 2014. View Article : Google Scholar
19	Kumfu S, Khamseekaew J, Palee S, Srichairatanakool S, Fucharoen S, Chattipakorn SC and Chattipakorn N: Combined iron chelator and T-type calcium channel blocker exerts greater efficacy on cardioprotection than monotherapy in iron-overload thalassemic mice. Eur J Pharmacol. 822:43–50. 2018. View Article : Google Scholar : PubMed/NCBI
20	Zhang Y, Zhao X, Chang Y, Zhang Y, Chu X, Zhang X, Liu Z, Guo H, Wang N, Gao Y, et al: Calcium channel blockers ameliorate iron overload-associated hepatic fibrosis by altering iron transport and stellate cell apoptosis. Toxicol Appl Pharmacol. 301:50–60. 2016. View Article : Google Scholar : PubMed/NCBI
21	Cui R, Choi SE, Kim TH, Lee HJ, Lee SJ, Kang Y, Jeon JY, Kim HJ and Lee KW: Iron overload by transferrin receptor protein 1 regulation plays an important role in palmitate-induced insulin resistance in human skeletal muscle cells. FASEB J. 12:fj201800448R2018.
22	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
23	Meyer JN, Leuthner TC and Luz AL: Mitochondrial fusion, fission, and mitochondrial toxicity. Toxicology. 391:42–53. 2017. View Article : Google Scholar : PubMed/NCBI
24	Wang H, Liu C, Zhao Y and Gao G: Mitochondria regulation in ferroptosis. Eur J Cell Biol. 99:1510582019. View Article : Google Scholar : PubMed/NCBI
25	Tilokani L, Nagashima S, Paupe V and Prudent J: Mitochondrial dynamics: Overview of molecular mechanisms. Essays Biochem. 62:341–360. 2018. View Article : Google Scholar : PubMed/NCBI
26	Jing X, Lin J, Du T, Jiang Z, Li T, Wang G, Liu X, Cui X and Sun K: Iron overload is associated with accelerated progression of osteoarthritis: The role of DMT1 mediated iron homeostasis. Front Cell Dev Biol. 8:5945092021. View Article : Google Scholar : PubMed/NCBI
27	Chung JY, Kim HS and Song J: Iron metabolism in diabetes-induced Alzheimer's disease: A focus on insulin resistance in the brain. Biometals. 31:705–714. 2018. View Article : Google Scholar : PubMed/NCBI
28	Gao M, Monian P, Pan Q, Zhang W, Xiang J and Jiang X: Ferroptosis is an autophagic cell death process. Cell Res. 26:1021–1032. 2016. View Article : Google Scholar : PubMed/NCBI
29	Nieuwenhuizen L, Schutgens RE, van Asbeck BS, Wenting MJ, van Veghel K, Roosendaal G, Biesma DH and Lafeber FP: Identification and expression of iron regulators in human synovium: Evidence for upregulation in haemophilic arthropathy compared to rheumatoid arthritis, osteoarthritis, and healthy controls. Haemophilia. 19:e218–e227. 2013. View Article : Google Scholar : PubMed/NCBI
30	Camacho A, Simao M, Ea HK, Cohen-Solal M, Richette P, Branco J and Cancela ML: Iron overload in a murine model of hereditary hemochromatosis is associated with accelerated progression of osteoarthritis under mechanical stress. Osteoarthritis Cartilage. 24:494–502. 2016. View Article : Google Scholar
31	Gammella E, Buratti P, Cairo G and Recalcati S: The transferrin receptor: The cellular iron gate. Metallomics. 9:1367–1375. 2017. View Article : Google Scholar : PubMed/NCBI
32	Chen MP, Cabantchik ZI, Chan S, Chan GC and Cheung YF: Iron overload and apoptosis of HL-1 cardiomyocytes: Effects of calcium channel blockade. PLoS One. 9:e1129152014. View Article : Google Scholar : PubMed/NCBI
33	Lopez de Figueroa P, Lotz MK, Blanco FJ and Carames B: Autophagy activation and protection from mitochondrial dysfunction in human chondrocytes. Arthritis Rheumatol. 67:966–976. 2015. View Article : Google Scholar : PubMed/NCBI
34	Zhang Z, Xu T, Chen J, Shao Z, Wang K, Yan Y, Wu C, Lin J, Wang H, Gao W, et al: Parkin-mediated mitophagy as a potential therapeutic target for intervertebral disc degeneration. Cell Death Dis. 9:9802018. View Article : Google Scholar : PubMed/NCBI
35	Tsuchiya M, Ichiseki T, Ueda S, Ueda Y, Shimazaki M, Kaneuji A and Kawahara N: Mitochondrial stress and redox failure in steroid-associated osteonecrosis. Int J Med Sci. 15:205–209. 2018. View Article : Google Scholar : PubMed/NCBI
36	Suwanjang W, Wu KL, Prachayasittikul S, Chetsawang B and Charngkaew K: Mitochondrial dynamics impairment in dexamethasone-treated neuronal cells. Neurochem Res. 44:1567–1581. 2019. View Article : Google Scholar : PubMed/NCBI
37	Anderson CP, Shen M, Eisenstein RS and Leibold EA: Mammalian iron metabolism and its control by iron regulatory proteins. Biochim Biophys Acta. 1823:1468–1483. 2012. View Article : Google Scholar : PubMed/NCBI
38	Esposito G, Vos M, Vilain S, Swerts J, Valadas JD, Van Meensel S, Schaap O and Verstreken P: Aconitase causes iron toxicity in Drosophila pink1 mutants. PLoS Genet. 9:e10034782013. View Article : Google Scholar : PubMed/NCBI
39	Bresgen N and Eckl PM: Oxidative stress and the homeodynamics of iron metabolism. Biomolecules. 5:808–847. 2015. View Article : Google Scholar : PubMed/NCBI