Melatonin Attenuates AlCl3-Induced Apoptosis and Osteoblastic Differentiation Suppression by Inhibiting Oxidative Stress in MC3T3-E1 Cells
Aluminum (Al) inhibits osteoblast-mediated bone formation by oxidative stress, resulting in Al-induced bone disease. Melatonin (MT) has received extensive attention due to its antioxidant and maintenance of bone health effect. To evaluate the protective effect and mechanism of MT on AlCl3-induced osteoblast dysfunction, MC3T3-E1 cells were treated with MT (100 μM) and/or AlCl3 (8 μM). First, MT alleviated AlCl3-induced osteoblast dysfunction, presenting as the reduced apoptosis rate as well as increased cell viability, alkaline phosphatase (ALP) activity, and type I collagen (COL-1) level. Then, MT significantly attenuated AlCl3-induced oxidative stress, presenting as the reduced reactive oxygen species and 8-hydroxy-2′-deoxyguanosine levels as well as increased glutathione level and superoxide dismutase activity. Finally, MT protected MC3T3-E1 cells against p53-dependent apoptosis and differentiation suppression, as assessed by Caspase-3 activity, protein levels of p53, Bcl-2-associated X protein (Bax), B cell lymphoma gene 2 (Bcl-2), cytosolic Cytochrome c, Runt-related transcription factor 2 (Runx2), and Osterix, as well as the mRNA levels of Bax, Bcl-2, Runx2, Osterix, ALP, and COL-1. Overall, our findings demonstrate MT attenuates AlCl3-induced apoptosis and osteoblastic differentiation suppression by inhibiting oxidative stress in MC3T3-E1 cells.
KeywordsAluminum chloride Melatonin Apoptosis Osteoblastic differentiation Oxidative stress p53
This work was supported by the National Natural Science Foundation Project (31872530), the “Young Talent” Project of Northeast Agricultural University (18QC44), the Earmarked Fund For China Agriculture Research System (CARS-35), and the Open Project Program of Northeastern Science Inspection Station, China Ministry of Agriculture Key Laboratory of Animal Pathogen Biology (DY201709).
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no competing interests.
- 1.Klotz K, Weistenhöfer W, Neff F, Hartwig A, Van TC, Drexler H (2017) The health effects of aluminum exposure. Dtsch Arztebl Int 114:653–659Google Scholar
- 3.Willhite CC, Karyakina NA, Yokel RA, Yenugadhati N, Wisniewski TM, Arnold IM, Momoli F, Krewski D (2014) Systematic review of potential health risks posed by pharmaceutical, occupational and consumer exposures to metallic and nanoscale aluminum, aluminum oxides, aluminum hydroxide and its soluble salts. Crit Rev Toxicol 44:1–80CrossRefGoogle Scholar
- 8.Krewski D, Yokel RA, Nieboer E, Borchelt D, Cohen J, Harry J, Kacew S, Lindsay J, Mahfouz AM, Rondeau V (2007) Human health risk assessment for aluminium, aluminium oxide, and aluminium hydroxide. J Toxicol Environ Health B Crit Rev 10:11–269Google Scholar
- 9.Natalia VD, Raja S, Jennifer LV, Shaoyu Z, Jason U, Richard N (2012) The metal transporter SMF-3/DMT-1 mediates aluminum-induced dopamine neuron degeneration. J Neurochem 124:147–157Google Scholar
- 14.Almeida M, Han L, Martin-Millan M, Plotkin LI, Stewart SA, Roberson PK, Kousteni S, O'Brien CA, Bellido T, Parfitt AM, Weinstein RS, Jilka RL, Manolagas SC (2007) Skeletal involution by age-associated oxidative stress and its acceleration by loss of sex steroids. J Biol Chem 282:27285–27297CrossRefGoogle Scholar
- 20.Li J, Zheng X, Ma X, Xu X, Du Y, Lv Q, Li X, Wu Y, Sun H, Yu L, Zhang Z (2019) Melatonin protects against chromium(VI)-induced cardiac injury via activating the AMPK/Nrf2 pathway. J Biol Chem 197:110698Google Scholar
- 21.Fischer TW, Kleszczynski K, Hardkop LH, Kruse N, Zillikens D (2013) Melatonin enhances antioxidative enzyme gene expression (CAT, GPx, SOD), prevents their UVR-induced depletion, and protects against the formation of DNA damage (8-hydroxy-2′-deoxyguanosine) in ex vivo human skin. J Pineal Res 54:303–312CrossRefGoogle Scholar
- 38.Xu F, Ren L, Song M, Shao B, Han Y, Cao Z, Li Y (2017) Fas- and mitochondria-mediated signaling pathway involved in osteoblast apoptosis induced by AlCl3. Biol Trace Elem Res 184:1–13Google Scholar
- 40.Kim HJ, Kim SH, Kim MS, Lee EJ, Oh HG, Oh WM, Park SW, Kim WJ, Lee GJ, Choi NG, Koh JT, Dinh DB, Hardin RR, Johnson K, Sylvia VL, Schmitz JP, Dean DD (2005) Varying Ti-6Al-4V surface roughness induces different early morphologic and molecular responses in MG63 osteoblast-like cells. J Biomed Mater Res A 74:366–373CrossRefGoogle Scholar
- 45.Miyashita T, Harigai M, Hanada M, Reed JC (1994) Identification of a p53-dependent negative response element in the bcl-2 gene. Cancer Res 54:3131–3135Google Scholar
- 49.Savitskaya MA, Onishchenko GE (2015) Mechanisms of apoptosis. Biochemistry 80:1393–1405Google Scholar