Lithium chloride increases differentiation and autophagy of osteoblasts inhibited by estrogen receptor

doi:10.3969/j.issn.2095-4344.1613

Abstract

Abstract:

BACKGROUND: Lithium has been used in the treatment of hematopoietic diseases and bipolar diseases for decades, and it can directly stimulate the proliferation of bone marrow mesenchymal stem cells.
OBJECTIVE: To investigate the effects of lithium chloride on the proliferation, differentiation and autophagy of rat osteoblasts inhibited by estrogen receptor.
METHODS: Ten neonatal Sprague-Dawley rats, 72 days old, SPF grade, were provided by the Heilongjiang University of Chinese Medicine. (1) Osteoblasts were isolated by enzymatic digestion and alizarin red staining, and logarithmically growing osteoblasts were then cultured in media containing different concentrations of lithium chloride (0, 1, 2, 5 nmol/L) for 24 hours. Cell proliferation was measured by cell counting kit-8. (2) Osteoblasts at logarithmical growth phase were randomly divided into blank group (normal cultured cells), inhibitor group (10^-7 mol/L ICI182780), treatment group (5 nmol/L lithium chloride+10^-7 mol/L ICI 182780). Calcified nodule staining and alkaline phosphatase activity were used to detect the differentiation of osteoblasts. Expression of Beclin1 and Runx2 in osteoblasts were determined by western blot.
RESULTS AND CONCLUSION: Under the inverted microscope, the cells were mostly mononuclear, polygonal and fusiform, with local cell-dense cell clusters. Results from the cell counting kit-8 assay showed that different concentrations of lithium chloride promoted the proliferation of osteoblasts in a concentration-dependent manner. Alizarin red staining results showed that the mineralization ability of osteoblasts was significantly reduced in the inhibitor group compared with the blank group, while compared with the inhibitor group, the mineralization ability of osteoblasts was increased significantly in the treatment group (P < 0.05). The activity of alkaline phosphatase was significantly reduced in the inhibitor group compared with the blank group, while compared with the inhibitory group, the activity of alkaline phosphatase increased significantly in the treatment group (P < 0.05). Western blot results showed that the expression of Beclin1 and Runx2 proteins in the inhibitor group decreased significantly compared with blank group, while compared with the inhibitor group, the expression of Beclin1 and Runx2 protein in the treatment group increased significantly (P < 0.05). In conclusion, lithium chloride can increase the proliferation, differentiation and autophagy of rat osteoblasts inhibited by estrogen receptor.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Lithium Chloride, Osteoblasts, Autophagy, Tissue Engineering

CLC Number:

R394.2

Fu Yin, Sun Guicai, Chen Shuilin, Fan Xiangwei, Peng Yufei. Lithium chloride increases differentiation and autophagy of osteoblasts inhibited by estrogen receptor[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(5): 680-684.

Figures/Tables 2

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