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Updated: Dec 30, 2025

A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation
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A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation

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Leptin inhibits AMPKα2 down-regulation induced decrease in the osteocytic MLO-Y4 cell proliferation and the

Qing Fan1, Hao Li1, Zhu Liu1

  • 1Department of Pediatric Orthopedics, Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine Shanghai, P. R. China.

International Journal of Clinical and Experimental Pathology
|January 23, 2020
PubMed

Insights

AMP-activated protein kinase alpha 2 (AMPKα2) knockdown inhibited osteocytic cell proliferation and osteogenic marker expression. Leptin treatment, however, enhanced these processes, suggesting AMPKα2

Area of Science:

  • Biochemistry
  • Cell Biology
  • Bone Biology

Background:

  • AMP-activated protein kinase (AMPK) regulates energy homeostasis.
  • AMPKα1 is involved in bone formation, but AMPKα2's role in osteogenesis is unclear.

Purpose of the Study:

  • To investigate the role of AMPKα2 in osteocytic MLO-Y4 cell proliferation and osteogenic marker expression.
  • To determine the effect of leptin on these processes in the presence or absence of AMPKα2.

Main Methods:

  • AMPKα2 was silenced in MLO-Y4 cells using pLKO.1-AMPKα2-shRNA.
  • Cell proliferation was assessed using CCK-8 assays.
  • Osteogenic marker expression (OPG, OCN, OPN, ALP, BMP6) was analyzed via Real-time PCR and Western blot.
  • Global transcriptomic changes were evaluated using RNA-seq.

Main Results:

  • AMPKα2 knockdown significantly reduced MLO-Y4 cell proliferation and expression of osteogenic markers (OPG, OCN, OPN, ALP, BMP6) and p-Smad5/Smad5.
  • Leptin treatment increased cell proliferation and osteogenic marker expression, irrespective of AMPKα2 levels.
  • RNA-seq revealed distinct transcriptional changes upon AMPKα2 silencing and leptin treatment.

Conclusions:

  • AMPKα2 plays a crucial role in promoting osteocytic proliferation and osteogenesis in vitro.
  • Leptin can counteract the inhibitory effects of AMPKα2 knockdown on osteogenesis.
Keywords:
AMPKα2GSEARNA-seqleptinosteogenesis

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