High glucose inhibits osteogenic differentiation and proliferation of MC3T3‑E1 cells by regulating P2X7

Yang,Jinsan; Ma,Cao; Zhang,Maoshu

doi:10.3892/mmr.2019.10790

High glucose inhibits osteogenic differentiation and proliferation of MC3T3‑E1 cells by regulating P2X7

Authors:
- Jinsan Yang
- Cao Ma
- Maoshu Zhang
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Affiliations: Department of Orthopedics, Jining No. 1 People's Hospital, Jining, Shandong 272011, P.R. China
Published online on: October 31, 2019 https://doi.org/10.3892/mmr.2019.10790
Pages: 5084-5090
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Diabetes mellitus adversely affects human bones and increases the risk of developing osteoporosis. In the present study, treatment with 30 mmol/l glucose was used to establish a high glucose (HG) cell model in vitro. Plasmids were used to overexpress the P2X purinoceptor 7 (P2X7) gene. Brilliant blue G and (4‑benzoyl‑benzoyl)‑ATP were used as a P2X7 antagonist and agonist, respectively. Proliferation of osteogenic MC3T3‑E1 cells and alkaline phosphatase (ALP) activity were determined using MTT and colorimetric assays, respectively. Alizarin Red S was used to assess calcification of MC3T3‑E1 cells. Western blotting and reverse transcription‑quantitative PCR were performed to determine protein and mRNA expression levels. The results demonstrated that HG inhibited MC3T3‑E1 cell proliferation and P2X7 expression, reduced calcification, and downregulated the expression levels of ALP and osteocalcin (Ocn) in MC3T3‑E1 cells. Overexpression of P2X7 in HG conditions increased calcification and proliferation, and upregulated the levels of ALP and Ocn in MC3T3‑E1 cells. Inhibition of P2X7 downregulated the expressions of ALP and Ocn in MC3T3‑E1 cells under HG conditions. Therefore, the present results indicated that HG caused damage to osteogenic MC3T3‑E1 cells. Thus, P2X7 may be a regulatory factor that may be used to counteract the effects of HG on osteogenesis.

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