2023, Vol. 27 ›› Issue (6): 878-882

Casein kinase 2-interaction protein-1 regulates the osteogenic ability of bone marrow mesenchymal stem cells in osteoporosis rats

Long Yanming1, Xie Mengsheng2, 3, Huang Jiajie2, 3, Xue Wenli2, 3, Rong Hui2, 3, Li Xiaojie2, 3   

Abstract: BACKGROUND: At present, in vitro studies on the molecular mechanism of casein kinase 2-interaction protein-1 (CKIP-1) mainly focus on gene knockout mouse-derived osteoblasts or bone marrow mesenchymal stem cells. There are few reports focusing on the expression of CKIP-1 in bone marrow mesenchymal stem cells derived from osteoporosis model rats.
OBJECTIVE: To investigate the changes in osteogenic differentiation of bone marrow mesenchymal stem cells before and after down-regulation of CKIP-1 gene.
METHODS: An osteoporotic rat model was induced by retinoic acid gavage. The bone marrow mesenchymal stem cells of the osteoporosis group and the normal group were cultured in vitro by the whole bone marrow adherence method. After osteogenic induction, Alizarin red staining, alkaline phosphatase staining and real-time quantitative RT-PCR were utilized to detect the relative expression of osteopontin and Runx2 mRNA. The dynamic expression of CKIP-1 in the two groups of cells was detected by using real-time quantitative RT-PCR. CKIP-1 gene was silenced by gene transfection. After osteogenic induction, alizarin red staining, alkaline phosphatase staining, and real-time quantitative RT-PCR were performed to detect the relative expression of osteopontin and Runx2 mRNA.
RESULTS AND CONCLUSION: (1) Compared with the normal group, alizarin red staining of calcium nodules, alkaline phosphatase activity and mRNA levels of osteopontin and Runx2 in bone marrow mesenchymal stem cells decreased in the osteoporosis group (P < 0.05). The dynamic expression level of CKIP-1 gene in bone marrow mesenchymal stem cells was generally higher in the osteoporosis group. (2) Compared with bone marrow mesenchymal stem cells of osteoporosis group without down-regulation of CKIP-1, after down-regulation of CKIP-1 gene expression, the quantification of calcium nodules and alkaline phosphatase activity by alizarin red staining, mRNA levels of osteopontin and Runx2 were significantly increased in bone marrow mesenchymal stem cells of the osteoporosis group (P < 0.05). (3) Therefore, osteogenic ability of bone marrow mesenchymal stem cells in rats with retinoic acid-induced osteoporosis is reduced and down-regulation of CKIP-1 gene can partially improve their osteogenic differentiation ability.
Key words: rat, retinoic acid, CKIP-1, osteoporosis, bone marrow mesenchymal stem cell, osteogenic differentiation