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[Effect of naringin combined with bone morphogenetic protein-2 on the proliferation and differentiation of MC3T3-E1 cells].

ABSTRACT: OBJECTIVE:This study evaluates the biological effects of naringin (NAR) joint bone morphogenetic protein (BMP)-2 on the proliferation, alkaline phosphatase (ALP) activity, and expression of osteoblastogenic genes, such as Runt-related transcription factor 2 (Runx2), collagen ? (Col?), ALP, and osteocalcin (OCN) of pre-osteoblasts. METHODS:Three different NAR concentrations (10, 100, and 1 000 ?mol·L?¹) were applied, alone or combined with BMP-2(50 ng·mL?¹), to restore the osteoblastogenesis of pre-osteoblasts (MC3T3-E1 cell line). Cell numbers (proliferation) were evaluated at first, fourth, and seventh days by Alamar blue assay. ALP activity and the expression of osteoblastogenic genes, such as Runx2, Col?, ALP, and OCN were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) at fourth and seventh day. RESULTS:Stimulation by NAR alone and in combination with BMP-2 for 1 day and 4 days could promote cell proliferation, which peaked at a concentration of 100 ?mol·L?¹ NAR combined with BMP-2 could promote cell proliferation significantly (P<0.05). Stimulation by NAR alone and in combination with BMP-2 for 4 and 7 days could promote ALP activity and bone-related gene(ALP, OCN, Runx2, Col?) expression. ALP expression was significantly promoted after stimulation of 100 ?mol·L?¹ NAR and BMP-2 (P<0.05). CONCLUSIONS:NAR exhibits promising potential for improving MC3T3-E1 proliferation and differentiation, and appropriate concentrations of NAR and BMP-2 show synergistic effect.?.

SUBMITTER: Gaoli X

PROVIDER: S-EPMC7030434 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature

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[Effect of naringin combined with bone morphogenetic protein-2 on the proliferation and differentiation of MC3T3-E1 cells].

Gaoli Xu X Yi Liu L Lili Wu W Qiutao Shi S Guang Huo H Zhiyuan Gu G

Hua xi kou qiang yi xue za zhi = Huaxi kouqiang yixue zazhi = West China journal of stomatology 20170601 3

<h4>Objective</h4>This study evaluates the biological effects of naringin (NAR) joint bone morphogenetic protein (BMP)-2 on the proliferation, alkaline phosphatase (ALP) activity, and expression of osteoblastogenic genes, such as Runt-related transcription factor 2 (Runx2), collagen Ⅰ (ColⅠ), ALP, and osteocalcin (OCN) of pre-osteoblasts.<h4>Methods</h4>Three different NAR concentrations (10, 100, and 1 000 μmol·L⁻¹) were applied, alone or combined with BMP-2(50 ng·mL⁻¹), to restore the osteobla ...[more]

PMID: 28675012

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Project description:BackgroundIn mouse, it was discovered that resveratrol (Res) enhanced osteoporosis (OP) by boosting osteogenesis. Besides, Res can also have an impact on MC3T3-E1 cells, which are crucial for the control of osteogenesis and thus increase osteogenesis. Although some articles have discovered that Res enhanced autophagy to promote the value-added differentiation of MC3T3, it is unclear exactly how this affects the process of osteogenesis in mouse. Therefore, we will show that Res encourages MC3T3-E1 proliferation and differentiation in mouse pre-osteoblasts and further investigate the autophagy-related mechanism for this impact.Methods(1) MC3T3-E1 cells were separated into blank control group and various concentrations (0.01, 0.1, 1, 10, 100µmol/L) of group in order to determine the ideal Res concentration. In the Res group, Cell Counting Kit-8 (CCK-8) was used to measure the proliferation activity of pre-osteoblasts in mice in each group after resveratrol intervention. Alkaline Phosphatase (ALP) and alizarin red staining were used to gauge the degree of osteogenic differentiation, and RT-qPCR was used to measure the expression levels of Runx2 and OCN in the osteogenic differentiation ability of the cells. (2) In the experiment, four groups were set up: the control group, 3MA group, Res group, and Res + 3MA group. To examine cell mineralization, ALP and alizarin red staining were utilized. RT-qPCR and Western blot detection of cell autophagy activity levels and osteogenic differentiation capacity in each group following intervention.Results(1) Resveratrol might increase the number of mice pre-osteoblast, with the impact being most pronounced at 10µmol/L (P < 0.05). The nodules developed substantially more often than in the blank control group, and Runx2 and OCN expressions significantly increased (P < 0.05). (2) In contrast to the Res group, after 3MA purine blocked autophagy, the Res + 3MA group's alkaline phosphatase staining and the development of mineralized nodules were reduced. Runx2, OCN, LC3II / LC3I expression decreased, p62 expression increased (P < 0.05).ConclusionThe present study partially or indirectly demonstrated that Res may, through increased autophagy, induce osteogenic differentiation of MC3T3-E1 cells.

Dataset Information

[Effect of naringin combined with bone morphogenetic protein-2 on the proliferation and differentiation of MC3T3-E1 cells].

Publications

[Effect of naringin combined with bone morphogenetic protein-2 on the proliferation and differentiation of MC3T3-E1 cells].

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