Chinese Medical Sciences Journal ›› 2016, Vol. 31 ›› Issue (2): 100-106.doi: 10.1016/S1001-9294(16)30033-5

• Orginal Article • Previous Articles     Next Articles

Mechanical Strain Regulates Osteoblast Proliferation Through Ca2+-CaMK-CREB Signal Pathway

Yong Guo1, 2, Qi Lv3, Xian-qiong Zou1, Zhi-xiong Yan1, Yu-xian Yan1, 3, *   

  1. 1Depantment of Bioengineering, College of Biotechnology, Guilin Medical University, Guilin, Guangxi 541004, China; 2Institute of Medical Equipment, Academy of Military Medical Sciences, Tianjin 300161, China; 3Experiment Management Center, Logistical College of People Armed Police Forces, Tianjin 300162, China
  • Received:2015-07-06 Online:2016-06-20 Published:2016-06-20
  • Contact: Tel:86-773-3680651 E-mail:2201642732@qq.com
  • Supported by:
    △Supported by the National Natural Science Foundation of China (11432016, 31370942, 11372351), and Higher School Science Foundation of Guangxi (04020150032)

Abstract: Objective To investigate the effects of mechanical strain on Ca2+-calmodulin dependent kinase (CaMK)-cAMP response element binding protein (CREB) signal pathway and proliferation of osteoblasts.Methods Using a four-point bending device, MC3T3-E1 cells were exposed to mechanical tensile strains of 2500 µs and 5000 µs at 0.5 Hz respectively. The intracellular free Ca2+ ([Ca2+]i) concentration and calmodulin activity were assayed by fluorospectrophotometry, CaMK II β, CREB, and phosphorylated (activated) CREB (p-CREB) were assessed by Western blot, and cells proliferation was assayed with MTT. Pretreatment with verapamil was carried out to block Ca2+ channel, and inhibitor U73122 was used to inhibit phospholipase C (PLC).Results Mechanical strains of 2500 µs and 5000 µs for 1 to 10 minutes both increased [Ca2+]i level of the cells. The 2500 µs strain, a periodicity of 1 h/d for 3 days, activated calmodulin, elevated protein levels of CaMK II β and p-CREB, and promoted cells proliferation, which were attenuated by pretreatment of verapamil or U73122. The effects of 5000 µs strain on calmodulin, CaMK II β, p-CREB and proliferation were contrary to 2500 µs strain.Conclusion The mechanical strain regulates osteoblasts proliferation through Ca2+-CaMK-CREB signal pathway via Ca2+ channel and PLC/IP3 transduction cascades.

Key words: mechanical strain, calcium, phospholipase C, proliferation, cAMP response element binding protein

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