LncRNA ZBED3-AS1/miR-339-5p/Notch 1 axis regulates osteoblast proliferation and differentiation in osteoporotic rats / 中华内分泌外科杂志

ABSTRACT

Objective:To investigate the role of LncRNA ZBED3-AS1 in osteoblast proliferation and differentiation in osteoporotic rats through regulating miR-339-5p/Notch 1.Methods:The rat models of sham operation (Sham) group and model (Model) group were established, and the bone mineral density (BMD) of rats was examined. Rat osteoblasts were isolated and the expression of ZBED3-AS1 and miR-339-5p was detected by qRT-PCR. The osteoblasts of rats in Sham group and Model group were divided into different groups and transfected. CCK8, alizarin red (AR-S) staining and alkaline phosphatase (ALP) staining were used to detect the proliferation and differentiation ability of cells in each group. The distribution of ZBED3-AS1 in cells was determined by FISH assay. Double luciferase report confirmed the relationship between ZBED3-AS1 and miR-339-5p as well as miR-339-5p and Notch 1.Western blot was used to detect the expression of Notch pathway related factors.Results:The bone mineral density of femur in Model group was significantly lower than that in Sham group ( P=0.0057) . Compared with Sham group, the expression of ZBED3-AS1 in osteoblasts of Model group was lower, while that of miR-339-5p was higher (all P<0.05) . Overexpression of ZBED3-AS1 could promote the proliferation and differentiation of osteoblasts, while knockdown of ZBED3-AS1 could inhibit the proliferation and differentiation of osteoblasts (all P<0.05) . ZBED3-AS1 could regulate miR-339-5p as ceRNA (all P<0.05) . Overexpression of miR-339-5p can inhibit the proliferation and differentiation of osteoblasts, which can be partially saved by overexpression of ZBED3-AS1. Notch 1 was confirmed as a target of miR-339-5p, at the same time, interfering with the expression of ZBED3-AS1/miR-339-5p can affect the expression of Notch-1 protein, and the regulation of ZBED3-AS1/miR-339-5p on osteoblasts may be realized through Notch pathway. Conclusion:ZBED3-AS1 can be used as ceRNA to regulate miR-339-5p, and then affect the proliferation and differentiation of osteoblasts in osteoporotic rats.