Effect of lithium chloride on AKT-mTOR signal pathway in osteoblasts after estrogen receptor inhibition

doi:10.3969/j.issn.2095-4344.1246

Abstract

Abstract:

BACKGROUND: Lithium chloride treats osteoporosis by activating Wnt signaling pathway, and can induce autophagy. Thereafter, improving the expression of lithium chloride in Akt and mTOR phosphorylation can promote osteoblast differentiation.
OBJECTIVE: To analyze the effect of lithium chloride on AKT-mTOR signal pathway in osteoblasts after estrogen receptor inhibition.
METHODS: Sprague-Dawley newly born 3-day rats, SPF grade (provided by Laboratory Animal Center of Heilongjiang University of Chinese Medicine) were selected. The osteoblasts were isolated from rat cranial bone by enzyme digestion, and then identified by alizarin red staining. The osteoblasts were randomized into blank control (normal culture), inhibitor (ICI182780), and treatment (lithium chloride plus ICI182780) groups. The cell morphology was observed under inverted microscope. Different concentrations of lithium chloride (0, 1, 2, 5 nmol/L) were added and cultured for 24 hours, and the cell proliferation was detected by cell counting-kit 8 assay. The osteoblast differentiation was detected by alizarin red staining and alkaline phosphatase activity. The contents of p-AKT and p-mTOR in osteoblasts were determined by western blot assay.
RESULTS AND CONCLUSION: (1) Inverted microscope showed that most of cells were mononuclear, polygonal and fusiform, with local cell-dense cell clusters. (2) Different concentrations of lithium chloride could promote osteoblast proliferation in a concentration-dependent manner. (3) Compared with the blank control group, mineralization ability of osteoblasts in the inhibitor was significantly decreased. Compared with the inhibitor group, the mineralization ability of osteoblasts in the treatment group was significantly increased. (4) The alkaline phosphatase activity in the inhibitor group was significantly lower than that in the blank control and treatment groups. (5) The expression levels of p-AKT and p-mTOR in the inhibitor group were significantly lower than those in the blank control and treatment groups. (6) To conclude, lithium chloride can promote the proliferation and differentiation of osteoblasts by activating the AKT-mTOR pathway with estrogen receptor inhibition.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: lithinm chloride, osteoblasts, estrogen receptor, osteoporosis, osteoblast autophagy, osteoblast differentiation

CLC Number:

R446

Xu Yier1, Sun Guicai2, Chen Shuilin2, Fan Xiangwei2 . Effect of lithium chloride on AKT-mTOR signal pathway in osteoblasts after estrogen receptor inhibition[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(19): 3002-3006.

Figures/Tables 2

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