Chinese Journal of Tissue Engineering Research ›› 2020, Vol. 24 ›› Issue (31): 4921-4926.doi: 10.3969/j.issn.2095-4344.2117

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Rev-erbα’s effect on osteoblastogenesis of mouse bone marrow mesenchymal stem cells 

Zhang Shuang1, 2, Xu Xiaomei1, 2, Zeng Yang1, 2, Yuan Xiaoping1, 2, Lin Fuwei1, 2    

  1. 1Department of Orthodontics, the Affiliated Stomatology Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China; 2Oral & Maxillofacial Reconstruction and Regeneration Laboratory, Southwest Medical University, Luzhou 646000, Sichuan Province, China

  • Received:2019-10-21 Revised:2019-10-24 Accepted:2019-12-26 Online:2020-11-08 Published:2020-09-03
  • Contact: Lin Fuwei, Master, Attending physician, Department of Orthodontics, the Affiliated Stomatology Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China; Oral & Maxillofacial Reconstruction and Regeneration Laboratory, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • About author:Zhang Shuang, Master candidate, Department of Orthodontics, the Affiliated Stomatology Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China; Oral & Maxillofacial Reconstruction and Regeneration Laboratory, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Supported by:
    the Project of Sichuan Medical Association, No. S18002; the Fund of Luzhou Municipal Department of Science and Technology, No. 2015-S-50(2/3) ; the Youth Foundation of the Affiliated Stomatological Hospital of Southwest Medical University, No. 201816

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Abstract:

BACKGROUND: The orphan nuclear receptor Rev-erbα has been demonstrated to play important roles during bone metabolism. However, the specific mechanism underlying osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) is still unclear.

OBJECTIVE: To explore whether the orphan nuclear receptor Rev-erbα participates in the osteogenesis process of BMSCs in mice.

METHODS: Mouse BMSCs were isolated and cultured by the whole bone marrow adherence method. And the lentivirus vector carrying the Rev-erbα gene was constructed and then transfected into the BMSCs. Real-time PCR was conducted to detect the mRNA level of Rev-erbα at 48 hours after transfection. There were three groups in the experiment: an experimental group with BMSCs transfected with lentiviral vectors containing overexpressed Rev-erbα gene and EGFP gene, a positive control group with BMSCs transfected with empty lentiviral vectors containing only EGFP gene, and a negative control group. Then the BMSCs were induced to differentiate into osteoblasts after the transfection and we detected the mRNA level of alkaline phosphatase, osteopontin, and osteocalcin using real-time PCR at 0, 7, and 14 days after induction.

RESULTS AND CONCLUSION: The lentivirus vector carrying Rev-erbα gene was successfully transfected into BMSCs, and expressed stably. An increase in the mRNA levels of alkaline phosphatase and osteopontin was detected in each group when cultured in osteogenic medium; however, there was no significant difference between groups. The mRNA level of osteocalcin was also increased in each group, and moreover, the mRNA level in the experimental group was significantly lower than that in the two control groups (P < 0.05). These results indicate that Rev-erbα transfection reduces the osteogenic ability of BMSCs, and the expression of osteocalcin, a late-stage osteogenic marker, is inhibited, indicating that the late-stage osteogenesis of BMSCs may be influenced by Rev-erbα.

Key words: bone marrow mesenchymal stem cells,  osteogenesis,  lentivirus,  Rev-erbα,  nuclear receptor

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