Abstract
Context: Sika pilose antler type I collagen (SPC-I) have been reported to promote bone marrow mesenchymal stem cell (BMSC) proliferation and differentiation. However, the underlying mechanism is still unclear.Objective: This study investigates the molecular mechanisms of SPC-I on the BMSC proliferation and differentiation of osteoblast (OB) in vitro.Material and methods: The primary rat BMSC was cultured and exposed to SPC-I at different concentrations (2.5, 5.0 and 10.0 mg/mL) for 20 days. The effect of SPC-I on the differentiation of BMSCs was evaluated through detecting the activity of alkaline phosphatase (ALP), ALP staining, collagen I (Col-I) content, and calcified nodules. The markers of osteoblastic differentiation were evaluated using RT-PCR and Western-blot analysis.Results: SPC-I treatment (2.5 mg/mL) significantly increased the proliferation of BMSCs (p < 0.01), whereas, SPC-I (5.0 and 10.0 mg/mL) significantly inhibited the proliferation of BMSCs (p < 0.01). SPC-I (2.5 mg/mL) significantly increased ALP activity and Col-I content (p < 0.01), and increased positive cells in ALP staining and the formation of calcified nodules. Additionally, the gene expression of ALP, Col-I, Osteocalcin (OC), Runx2, Osterix (Osx), ERK1/2, BMP2 and p38-MAPK, along with the protein expression of ERK1/2, p-ERK1/2, p-p38 MAPK were markedly increased in the SPC-I (5.0 mg/mL) treatment group (p < 0.01) compared to the control group.Discussion and conclusions: SPC-I can induce BMSC differentiation into OBs and enhance the function of osteogenesis through ERK1/2 and p38-MAPK signal transduction pathways and regulating the gene expression of osteogenesis-specific transcription factors.
Highlights
Osteoporosis, one of the most common bone diseases characterized by bone microstructure deterioration in addition to a reduction in bone mass, becomes a serious threat to post-menopausal women and elderly men (Lodie et al 2002; Meier et al 2005)
We proved the hypothesis that there was a cross-talk mechanism between ERK1/2 and p38-mitogenactivated protein kinases (MAPKs) that SPC-I acted on the differentiation and proliferation of bone marrow mesenchymal stem cell (BMSC)
We found that SPC-I promoted BMSC differentiation into osteoblast through ERK1/2 and p38-MAPK signal transduction pathways in addition to bone morphogenetic protein (BMP)-2/Smad signalling pathway
Summary
Osteoporosis, one of the most common bone diseases characterized by bone microstructure deterioration in addition to a reduction in bone mass, becomes a serious threat to post-menopausal women and elderly men (Lodie et al 2002; Meier et al 2005). The development of traditional Chinese medicine (TCM) has become a research topic of interest, and many natural medicines have been reported for use in the treatment of osteoporosis (Li et al 2003). Traditional use of antler velvet as medicines has been extensively recognized. The deficiency in kidney function is deemed to be closely related to osteoporosis among bones and joints (Okazaki and Sandell 2004; Bauer et al 2009). The velvet and its extracts are commonly administered to prevent and treat kidney deficiency related to osteoporosis in order to strengthen bone and muscles, reduce chronic joint pain (Wu et al 2013). Velvet antler components are the same as human bone having a calcium phosphate matrix (73%) with other organic material (23%). The Sika pilose antler type I collagen (SPC-I) (80%) is mainly made of fibrous tissues
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