The Korean Journal of Physiology and Pharmacology

The Korean Society of Pharmacology (대한약리학회)
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Nitric Oxide-Induced Autophagy in MC3T3-E1 Cells is Associated with Cytoprotection via AMPK Activation

  • Yang, Jung Yoon (Department of Oral Physiology, Dental Science Research Institute, School of Dentistry, Chonnam National University) ;
  • Park, Min Young (Department of Oral Physiology, Dental Science Research Institute, School of Dentistry, Chonnam National University) ;
  • Park, Sam Young (Department of Oral Physiology, Dental Science Research Institute, School of Dentistry, Chonnam National University) ;
  • Yoo, Hong Il (Department of Oral Anatomy, Dental Science Research Institute, School of Dentistry, Chonnam National University) ;
  • Kim, Min Seok (Department of Oral Anatomy, Dental Science Research Institute, School of Dentistry, Chonnam National University) ;
  • Kim, Jae Hyung (Department of Oral Medicine, Dental Science Research Institute, School of Dentistry, Chonnam National University) ;
  • Kim, Won Jae (Department of Oral Physiology, Dental Science Research Institute, School of Dentistry, Chonnam National University) ;
  • Jung, Ji Yeon (Department of Oral Physiology, Dental Science Research Institute, School of Dentistry, Chonnam National University)
  • Received : 2015.04.16
  • Accepted : 2015.08.07
  • Published : 2015.11.01
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Abstract

Nitric oxide (NO) is important in the regulation of bone remodeling, whereas high concentration of NO promotes cell death of osteoblast. However, it is not clear yet whether NO-induced autophagy is implicated in cell death or survival of osteoblast. The present study is aimed to examine the role of NO-induced autophagy in the MC3T3-E1 cells and their underlying molecular mechanism. The effect of sodium nitroprusside (SNP), an NO donor, on the cytotoxicity of the MC3T3-E1 cells was determined by MTT assay and expression of apoptosis or autophagy associated molecules was evaluated by western blot analysis. The morphological observation of autophagy and apoptosis by acridine orange stain and TUNEL assay were performed, respectively. Treatment of SNP decreased the cell viability of the MC3T3-E1 cells in dose- and time-dependent manner. SNP increased expression levels of p62, ATG7, Beclin-1 and LC3-II, as typical autophagic markers and augmented acidic autophagolysosomal vacuoles, detected by acridine orange staining. However, pretreatment with 3-methyladenine (3MA), the specific inhibitor for autophagy, decreased cell viability, whereas increased the cleavage of PARP and caspase-3 in the SNP-treated MC3T3-E1 cells. AMP-activated protein kinase (AMPK), a major autophagy regulatory kinase, was activated in SNP-treated MC3T3-E1 cells. In addition, pretreatment with compound C, an inhibitor of AMPK, decreased cell viability, whereas increased the number of apoptotic cells, cleaved PARP and caspase-3 levels compared to those of SNP-treated MC3T3-E1 cells. Taken together, it is speculated that NO-induced autophagy functions as a survival mechanism via AMPK activation against apoptosis in the MC3T3-E1 cells.

Keywords

References

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