Molecular mechanism by which Jiangu granules promote osteoblast proliferation

doi:10.3969/j.issn.2095-4344.2013.15.003

Abstract

Abstract:

BACKGROUND: Extracellular signal-regulated kinase is a protein kinase dependent on Ras pathway activation, and plays an important role in the proliferation and differentiation of osteoblasts. PD98059 is a specific MEK inhibitor, and inhibits phosphorylation of extracellular signal-regulated kinase through inhibition of MEK activity, thus blocking extracellular signal-regulated kinase signaling pathway. At present, the role of extracellular signal-regulated kinase in the process of proliferation and differentiation of rat osteoblasts is reported less. It is still unclear about the role of extracellular signal-regulated kinase in the regulation of osteoblast proliferation and differentiation promoted by Jiangu granules.
OBJECTIVE: To observe the role of extracellular signal-regulated kinase signaling pathway in the promotion of osteoblast proliferation and differentiation after intervention with Jiangu granules.
METHODS: Passage 3 osteoblasts from the skull of Sprague-Dawley rats were harvested and divided into four groups. Control group was intervened with saline serum. Drug group was intervened with the best concentration of serum containing Jiangu granules. Inhibitor group was intervened by PD98059. Combination group was intervened with PD98059 plus serum containing Jiangu granules. 3-(4,5-Dimethylthiazol-2- yl)-2,5- diphenyltetrazolium bromide assay was used to determine proliferative capacity of osteoblasts. Alkaline phosphatase and hydroxyproline levels were measured by colorimetric method. Expression of core binding factor alpha 1, type Ⅰ collagen, Osterix mRNA was measured by PCR-SYBR GREEN. Expression of extracellular signal-regulated kinase in osteoblasts was detected using western blot method.
RESULTS AND CONCLUSION: After blocking extracellular signal-regulated kinase signal pathway, the expression of extracellular signal-regulated kinase in the inhibitor and combination groups was significantly lower than that in the control and drug groups. Levels of alkaline phosphatase and hydroxyproline, as well as core binding factor alpha 1, type Ⅰ collagen, Osterix mRNA expression in osteoblasts, in the inhibitor group were significantly lower than those in the control and drug groups. It is indicated that extracellular signal-regulated kinase exerts an important role in promoting osteoblast proliferation and differentiation after intervention with Jiangu granules.

Key words: tissue construction, bone tissue construction, extracellular signal-regulated kinase, PD98059, Jiangu granules, osteoblasts, proliferation, differentiation, alkaline phosphatase, hydroxyproline, core binding factor alpha 1, type Ⅰ collagen, Osterix, National Natural Science Foundation of China

CLC Number:

R318

Lin Yu, Lu Tian-xiang, Wu Yin-sheng, Huang Yun-mei, Lin Yan-ping. Molecular mechanism by which Jiangu granules promote osteoblast proliferation[J]. Chinese Journal of Tissue Engineering Research, doi: 10.3969/j.issn.2095-4344.2013.15.003.

Figures/Tables 6

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