关键词: 骨碎补总黄酮, 成骨细胞, 增殖, miR-93-5p, WNT通路

Abstract:

Objective To investigate the effects of total flavonoids of rhizomadrynariae(TFRD)on the proliferation and apoptosis of alveolar bone osteoblasts and its mechanism. Methods Osteoblasts from rat alveolar bone were isolated and cultured. The cells were divided into control group, low-dose TFRD group(50 mg/L)and high-dose TFRD group(100 mg/L). After transfection with NC mimic and miR-93-5p mimic, osteoblasts were incubated with high-dose TFRD, which were classified as TFRD+NC mimic group and TFRD+ miR-93-5p mimic group, respectively. Cell proliferation was detected by CCK-8 method. Cell apoptosis was detected by flow cytometry. The expression of miR-93-5p, osteogenic differentiation-related genes(Osteriex and RUNX2), and WNT pathway-related genes(Dishevelled and β-catenin)were detected by RT-qPCR and Western blotting, respectively. Osteogenic differentiation was detected by ALP staining and activity test. Results After TFRD treatment, the proliferation ability of alveolar osteoblasts was increased(P<0.05); apoptosis rate was decreased(P<0.05); ALP activity and the expression of Osteriex and RUNX2 was increased(P<0.05); the expression of miR-93-5p was decreased(P<0.05); the expression of FZD6, Dishevelled and β-catenin was increased(P<0.05), activating the WNT pathway. Overexpression of miR-93-5p reversed the protective effect of TFRD on osteoblasts, leading to the reduction of cell proliferation(P<0.05), the increase of apoptosis rate(P<0.05), the decrease of ALP activity and Osteriex and RUNX2 expression(P<0.05), the increase of miR-93-5p expression(P<0.05), the reduction of FZD6, Disevelled and β-catenin expression(P<0.05), and the inhibition of WNT pathway. Conclusion TFRD promotes proliferation and inhibits apoptosis of alveolar bone osteoblasts through miR-93-5p/FZD6/WNT pathway.

Key words: total flavonoids of rhizomadrynariae, osteoblast, proliferation, miR-93-5p, WNT pathway