Mechanism analysis of ω-3 polyunsaturated fatty acids in alleviating oxidative stress and promoting osteogenic differentiation of MC3T3-E1 cells through activating Nrf2/NQO1 pathway.
10.7507/1002-1892.202506037
- Author:
Jiahui HUANG
1
;
Long CHEN
1
;
Chen XU
1
;
Haojie YU
1
;
Shishuai ZHOU
1
;
Jianzhong GUAN
1
Author Information
1. Department of Orthopaedics, the First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui, 233000, P. R. China.
- Publication Type:Journal Article
- Keywords:
MC3T3-E1 cells;
NAD (P) H quinone oxidoreductase 1;
nuclear factor E2-related factor 2;
oxidative stress;
ω-3 polyunsaturated fatty acids
- MeSH:
Oxidative Stress/drug effects*;
NF-E2-Related Factor 2/metabolism*;
NAD(P)H Dehydrogenase (Quinone)/metabolism*;
Animals;
Mice;
Osteogenesis/drug effects*;
Cell Differentiation/drug effects*;
Fatty Acids, Omega-3/pharmacology*;
Signal Transduction/drug effects*;
Osteoblasts/drug effects*;
Reactive Oxygen Species/metabolism*;
Cell Line;
Hydrogen Peroxide/pharmacology*;
Core Binding Factor Alpha 1 Subunit/metabolism*;
Antioxidants/pharmacology*;
Heme Oxygenase-1/metabolism*
- From:
Chinese Journal of Reparative and Reconstructive Surgery
2025;39(11):1459-1467
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To explore the mechanism by which ω-3 polyunsaturated fatty acids (hereinafter referred to as "ω-3") exert antioxidant stress protection and promote osteogenic differentiation in MC3T3-E1 cells, and to reveal the relationship between ω-3 and the key antioxidant stress pathway involving nuclear factor E2-related factor 2 (Nrf2) and NAD (P) H quinone oxidoreductase 1 (NQO1) in MC3T3-E1 cells.
METHODS:The optimal concentration of H 2O 2 (used to establish the oxidative stress model of MC3T3-E1 cells in vitro) and the optimal intervention concentrations of ω-3 were screened by cell counting kit 8. MC3T3-E1 cells were divided into blank control group, oxidative stress group (H 2O 2), low-dose ω-3 group (H 2O 2+low-dose ω-3), and high-dose ω-3 group (H 2O 2+high-dose ω-3). After osteoblastic differentiation for 7 or 14 days, the intracellular reactive oxygen species (ROS) level was measured by fluorescence staining and flow cytometry, and the mitochondrial morphological changes were observed by biological transmission electron microscope; the expression levels of Nrf2, NQO1, heme oxygenase 1 (HO-1), Mitofusin 1 (Mfn1), and Mfn2 were detected by Western blot to evaluate the cells' antioxidant stress capacity; the expression levels of Runt-related transcription factor 2 (RUNX2) and osteocalcin (OCN) were detected by immunofluorescence staining and Western blot; osteogenic potential of MC3T3-E1 cells was evaluated by alkaline phosphatase (ALP) staining and alizarin red staining.
RESULTS:Compared with the oxidative stress group, the content of ROS in the low and high dose ω-3 groups significantly decreased, and the protein expressions of Nrf2, NQO1, and HO-1 significantly increased ( P<0.05). At the same time, the mitochondrial morphology of MC3T3-E1 cells improved, and the expressions of mitochondrial morphology-related proteins Mfn1 and Mfn2 significantly increased ( P<0.05). ALP staining and alizarin red staining showed that the low-dose and high-dose ω-3 groups showed stronger osteogenic ability, and the expressions of osteogenesis-related proteins RUNX2 and OCN significantly increased ( P<0.05). And the above results showed a dose-dependence in the two ω-3 treatment groups ( P<0.05).
CONCLUSION:ω-3 can enhance the antioxidant capacity of MC3T3-E1 cells under oxidative stress conditions and upregulate their osteogenic activity, possibly through the Nrf2/NQO1 signaling pathway.
