Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (23): 4211-4214.doi: 10.3969/j.issn.1673-8225.2011.23.008

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Effect of receptor activator of nuclear factor kappa B ligand expression suppressed by RNA interference on osteogenic and adipogenic differentiation potential of bone marrow derived stroma cells 

Bai Yun, Yin Guo-zhu, Zhang Chao-liang, Zhang Xiao-hui, Luo En   

  1. State Key Laboratory of Oral Diseases, Sichuan University, Chengdu  610041, Sichuan Province, China
  • Received:2010-11-26 Revised:2011-03-20 Online:2011-06-04 Published:2011-06-04
  • Contact: Luo En, Doctor, Professor, State Key Laboratory of Oral Diseases, West China College of Stomatology, Sichuan University, Chengdu 610041, Sichuan Province, China donow3000@scu.edu.cn
  • About author:Bai Yun★, Studying for master’s degree, State Key Laboratory of Oral Diseases, West China College of Stomatology, Sichuan University, Chengdu 610041, Sichuan Province, China bai0caifly@126.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30973346*, 30700950*; Program for New Century Excellent Talents in Vniversity (NCET-10-0597)*

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Abstract:

BACKGROUND: RNA interference (RNAi) is an important method to down-regulate the gene expression in molecular biological study. Osteoprotegerin (OPG)/receptor activator of nuclear factor kappa B (RANK)/RANK ligand (RANKL) couple system is a hotspot in the research of bone remolding.
OBJECTIVE: To investigate the effect of RANKL expression suppressed by RNAi on osteogenic and adipogenic differentiation potential of bone marrow derived stroma cells (BMSCs).
METHODS: BMSCs were isolated and cultured from SD rats, then were seeded in 6-holes culture plates by 5 × 105 /hole 24 hours before transfection and divided into experimental group, negative control group and blank group. siRNA was transfected into the cells by Lipofectin 2000TM in experimental group and negative control group. Proliferation activity of BMSCs and alkaline phosphatase activity were measured, and the mRNA expression revolving in osteogenic and adipogenic differentiation of BMSCs were detected by real-time PCR.
RESULTS AND CONCLUSION: Alkaline phosphatase activity, RunX-2, bone morphogenetic protein-2 (BMP-2), and BMP-4 expression decreased, while peroxisome proliferator-activated receptor-γ and recombinant Human CCAAT/enhancer binding protein alpha expression increased after RANKL expression inhibited by RNAi (P < 0.05). The results suggest that the down-regulation of RANKL by RNAi could improve adipogenic differentiation and depress osteogenic differentiation of BMSCs.

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