Calcified Tissue International

, Volume 104, Issue 1, pp 59–69 | Cite as

Metformin Alleviates the Bone Loss Induced by Ketogenic Diet: An In Vivo Study in Mice

  • Qi Liu
  • Xiaolin Xu
  • Zhou Yang
  • Yapu Liu
  • Xiuhua Wu
  • Zhiping Huang
  • Junhao Liu
  • Zucheng Huang
  • Ganggang Kong
  • Jianyang Ding
  • Rong Li
  • Junyu Lin
  • Qingan ZhuEmail author
Original Research


Metformin (Met), an anti-diabetes drug, has also shown therapeutic effects for ovariectomy-induced (OVX) osteoporosis. However, similar effects against bone loss induced by a ketogenic diet (KD) have not been tested. This study was aimed to evaluate the microarchitectures and biomechanics of KD-induced osteoporosis with and without administration of Met, and compare the effect of Met on bone loss induced by KD with OVX. Forty female C57BL/6J mice were randomly divided into Sham, OVX, OVX + Met (100 mg/kg/day), KD (3:1 ratio of fat to carbohydrate and protein), and KD + Met (100 mg/kg/day) groups. After 12 weeks, the bone mass and biomechanics were measured in distal cancellous bone and in mid-shaft cortical bone of the femur. The activities of serum alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP), together with immunohistochemistry staining of osteocalcin (OCN) and TRAP, were evaluated. Both OVX and KD induced significant bone loss and compromised biomechanical properties in the cancellous bone, but no effect was found in cortical bone. The administration of Met increased the cancellous bone volume fraction (BV/TV) from 3.78 to 5.23% following the OVX and from 4.04 to 6.33% following the KD, it also enhanced the compressive stiffness from 47 to 160 N/mm following the OVX and from 35 to 340 N/mm with the KD. Met effectively increased serum ALP in the KD group while decreased serum TRAP in the OVX group, but up-regulated expression of OCN and down-regulated expression of TRAP in both OVX and KD groups. The present study demonstrated that Met effectively attenuated the cancellous bone loss induced by KD and maintained the biomechanical properties of long bones, providing evidence for Met as a treatment of by KD-induced osteoporosis in teenage skeleton.


Metformin Osteoporosis Ovariectomy Ketogenic diet Bone loss Micro-CT 



This study was supported by Guangdong Province Natural Science Foundation of China (No. 2015A030313276) and Dean Foundation of Nanfang Hospital (No. 2016Z021).

Author Contributions

QZ and QL designed the experiments. QL, ZY, and XX conducted the animal experiments. QL, ZY, JL, and YL collected the samples. QL, YZ, XX, XW and ZH measured and collected the data. ZH, GK, JD, RL and JL completed the data analysis. QL wrote the manuscript, and XX and QZ revised the manuscript.

Compliance with Ethical Standards

Conflict of interest

Qi Liu, Xiaolin Xu, Zhou Yang, Yapu Liu, Xiuhua Wu, Zhiping Huang, Junhao Liu, Zucheng Huang, Ganggang Kong, Jianyang Ding, Rong Li, Junyu Lin, and Qingan Zhu declare no conflicts of interest.

Human and Animal Rights and Informed Consent

The present study was approved by the Animal Experiments Ethics Committee of Southern Medical University. The animal procedures were conducted in accordance with the Guidelines of Caring for Laboratory Animals by the Ministry of Science and Technology of the People’s Republic of China. Surgery was performed under anesthesia, and all efforts were made to minimize suffering.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Qi Liu
    • 1
  • Xiaolin Xu
    • 1
  • Zhou Yang
    • 1
  • Yapu Liu
    • 1
    • 2
  • Xiuhua Wu
    • 1
  • Zhiping Huang
    • 1
  • Junhao Liu
    • 1
  • Zucheng Huang
    • 1
  • Ganggang Kong
    • 1
  • Jianyang Ding
    • 1
  • Rong Li
    • 1
  • Junyu Lin
    • 1
  • Qingan Zhu
    • 1
    Email author
  1. 1.Department of Spinal Surgery, Nanfang HospitalSouthern Medical UniversityGuangzhouChina
  2. 2.Department of Spinal SurgeryThe Second Affiliated Hospital of Luohe Medical CollegeLuoheChina

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