Pterostilbene promotes the osteogenic differentiation of MC3T3-E1 cells inhibited by H2O2 by activating the AMPK signaling pathways

Haiyan Li; Jiang Guo; Dong Zhang; Jihong Huang; Xuejia Li

doi:10.15586/qas.v14i1.1010

Haiyan Li

Department of Endocrinology, Shenzhen Second People’s Hospital (the First Affiliated Hospital of Shenzhen University), Shenzhen, Guangdong Province, China.

Jiang Guo

Department of Ophthalmology, Shenzhen Second People’s Hospital (the First Affiliated Hospital of Shenzhen University), Shenzhen, Guangdong Province, China.

Dong Zhang

Department of Orthopedics, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, China.

Jihong Huang

Department of Orthopedics, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, China.

Xuejia Li

Department of Orthopedics, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, China.

Keywords

pterostilbene, osteogenic differentiation, osteoporosis, AMPK pathway, MC3T3-E1

Abstract

Osteoporosis is one of the common degenerative skeletal diseases whose essential mechanism is the imbalance between bone resorption and formation. Currently, the close coordination of signaling pathways at genetic level during bone development is a hot topic of osteoporosis research. The present study is focused on whether pterostilbene protects against hydrogen peroxide (H₂O₂)-induced osteoblastic cell apoptosis, oxidative stress, and reveals the related underlying mechanisms. MC3T3-E1 osteoblastic cells were cultured and treated with different concentrations of H₂O₂ (0–1 mM) along with or without pterostilbene, and MTT assay or Annexin V-FITC/propidium iodide staining was applied for measuring cell viability and apoptosis. The in vitro cellular antioxidant analysis was performed using 2,7-dichlorodihydrofluorescein diacetate assay and enzyme-linked-immunosorbent serologic assay against glutathione peroxidase and malondialdehyde. In addition, cellular alkaline phosphatase activity was executed by Alizarin Red S staining. Western blot assay was conducted to determine the expression levels of osteogenic-related markers, including type I collagen, osteopontin, runt-related transcription factor 2, and the 5' adenosine monophosphate-activated protein kinase (AMPK) signaling pathways related proteins. The key finding was that pterostilbene could attenuate the H₂O₂-induced cellular apoptosis and oxidative stress. Pterostilbene also promoted osteogenic differentiation in H₂O₂-treated MC3T3-E1 cells through activation of the AMPK pathway. In conclusion, pterostilbene blocked the H₂O₂-induced MC3T3-E1 cells dysfunction, indicating its potential to be a promising medication for treating osteoporosis.

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