Bushen Huoxue Decoction promotes osteoporotic fracture healing in rats through Runx2/Osterix

doi:10.3969/j.issn.2095-4344.1093

Abstract

Abstract:

BACKGROUND: Runx2/Osterix is an important transcription factor promoting osteoblast differentiation. Chinese herbs have been shown to promote fracture healing through up-regulation of Runx2/Osterix. Bushen Huoxue Decoction is a prescription used to treat fractures and it can promote bone formation. Exploring underlying mechanism is helpful for its clinical application.
OBJECTIVE: To study the effect of Bushen Huoxue Decoction on osteoporotic fracture healing in rats and the underlying molecular mechanism.
METHODS: Forty 4-month-old adult female Sprague-Dawley rats provided by Laboratory Animal Centre of Guangzhou University of Chinese Medicine were randomly divided into five groups: model, Bushen Huoxue Decoction, Runx2 gene silencing, Runx2 gene silencing + Bushen Huoxue Decoction, and control groups. The femoral fracture model was established in the control group, while the osteoporotic femoral fracture model was established by ovariectomy in the other groups. Rats in the Bushen Huoxue Decoction and Runx2 gene silencing + Bushen Huoxue Decoction groups were treated with Bushen Huoxue Decoction, while rats in the model, control and Runx2 gene silencing groups were treated with equal volume of distilled water. Rats in the Runx2 gene silencing and Runx2 gene silencing + Bushen Huoxue Decoction groups were injected with Runx2 recombinant lentivirus, while rats in the model, control, and Bushen Huoxue Decoction groups were injected with equal volume of PBS at the same site. After 4 weeks, bone mineral density, tissue mineral density and bone volume fraction were measured by Micro CT. The mRNA and protein expression levels of the callus osteogenesis markers Runx2 and Osterix were detected by qPCR and western blot assay, respectively.
RESULTS AND CONCLUSION: Compared with the control group, bone mineral density, tissue mineral density, and expression levels of Runx2 and Osterix mRNA and protein in the model group were significantly decreased (P < 0.01). Compared with the model group, bone volume fraction and Runx2 and Osterix mRNA expression levels were significantly increased (P < 0.05), and Runx2 and Osterix protein expression levels were significantly increased (P < 0.01) in the Bushen Huoxue Decoction group. Compared with the model group, Osterix mRNA expression level and bone volume fraction were significantly decreased (P < 0.05), bone mineral density, tissue mineral density, Runx2 mRNA and protein expression levels, and Osterix protein expression level were significantly decreased (P < 0.01) in the Runx2 gene silencing group. Compared with the Runx2 gene silencing group, Runx2 and Osterix mRNA and protein expression levels, bone mineral density, tissue mineral density and bone volume fraction in the Runx2 gene silencing + Bushen Huoxue Decoction group were significantly increased (P < 0.01). To conclude, lentivirus-mediated Runx2 gene silencing can delay the healing of osteoporotic fracture in rats. Bushen Huoxue Decoction can up-regulate the expression of Runx2/Osterix and reverse the effect of Runx2 gene silencing, which can obviously promote the healing of fracture and has good therapeutic effect on osteoporotic fracture.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Tissue Engineering, Osteoporosis, Gene Silencing, Fracture Healing

CLC Number:

R453
R364

Hu Liuchao1, Luo Yiwen2, Cheng Yingxiong2, Wu Zhifang2, Luo Hui1, Shen Wei1. Bushen Huoxue Decoction promotes osteoporotic fracture healing in rats through Runx2/Osterix[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(11): 1717-1722.

Figures/Tables 3

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