Abstract
Purpose: To evaluate the effect of Eucommia ulmoides extract (EUE) on osteoblast proliferation as well as investigate its probable mechanisms of action. Methods: EUE was pharmacologically evaluated at three doses. Osteoblast cells were divided as follows: Group I: negative control; groups II–IV: received EUE (180, 360 and 540 μg/ml, respectively). We performed 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay to determine osteoblast viability following treatment. Alkaline phosphatase (ALP), osteocalcin, and collagen I levels in osteoblasts were quantified using commercially available kits. Thereafter, mRNA and protein expression of ALP, collagen I, osteocalcin, transforming growth factor-β1 (TGF-β1) were measured using real-time quantitative PCR (qPCR) and western blot, respectively. Results : EUE significantly (p < 0.01) promoted osteoblast proliferation at three treatment doses (180, 360, and 540 μg/mL). Furthermore, ALP, osteocalcin, collagen I and TGF-β1 expression at both mRNA and protein levels increased significantly (all p < 0.05) following EUE treatment. Conclusion : The results suggest that EUE may promote osteoblast cell proliferation and that ALP, osteocalcin, collagen I and TGF-β1 gene expression may be involved in the mechanism of action. Keywords: qPCR, collagen I, Bone, Liver, Eucommia ulmoides extract
Highlights
Osteoporosis reduces bone mass by destroying bone microstructure, leading to increased bone fragility
The effects of Eucommia ulmoides extract on cell viability and Alkaline phosphatase (ALP), collagen I, osteocalcin, and transforming growth factor-β1 (TGF-β1) mRNA and protein levels in osteoblasts were evaluated
ALP, osteocalcin and collagen I levels in osteoblasts increased significantly following treatment with Eucommia ulmoides extract (Table 3)
Summary
Osteoporosis reduces bone mass by destroying bone microstructure, leading to increased bone fragility. Eucommia ulmoides is a perennial deciduous tree that is endemic to China and protected by the government. This plant is often used for medicinal purposes. Eucommia ulmoides has been used to treat numbness, certain types of liver deficiencies, tinea cruris, and other conditions. The effects of Eucommia ulmoides extract on cell viability and ALP, collagen I, osteocalcin, and TGF-β1 mRNA and protein levels in osteoblasts were evaluated. We collected Eucommia ulmoides plants from XiangYang City, HuBei Province, China in 2015. Eucommia ulmoides extract was prepared in our laboratory as follows. Osteoblast were treated with extract for 48 h, washed with cold PBS three times, and lysed in RIPA buffer for 30 min on ice. Samples were centrifuged (12000 rpm, 4 °C, 20 min).
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