EFFECT OF PERIOSTIN ON AUTOPHAGY AND FACTORS RELATED TO BONE METABOLISM IN OSTEOBLASTS

Abstract

The purpose of this study was to explore the effect of periostin on autophagy and factors related to bone metabolism in osteoblasts. According to experimental design, some osteoblast-like cells (MC3T3-E1) were transfected by lentiviral vector to silence periostin gene. Celigo[Formula: see text] Image Cytometer and Real-Time PCR were used to assess target gene silencing effects. Western blotting was used to evaluate autophagy-associated factors (LC3B,Beclin1) and factors related to bone metabolism (RANKL, OPG, NF-[Formula: see text]B[Formula: see text]P65). Another part of osteoblasts was added with recombinant periostin protein at concentration of 75[Formula: see text]ng/mL. Monodansylcadaverine (MDC) detection was conducted to trace formation of autophagy vesicles. Factors associated with autophagy and bone metabolism were reassessed using western blotting. When periostin gene was silenced, western blotting analysis showed that the expression of LC3B-I, Beclin1, RANKL and NF-[Formula: see text]B[Formula: see text]P65 in experimental group was significantly increased; however, OPG expression was decreased. After adding recombinant periostin protein, MDC detection revealed significantly fewer green fluorescent particles in the experimental group. Western blotting results showed that the expression of LC3B, Beclin1, RANKL and NF-[Formula: see text]B[Formula: see text]P65 was decreased; however, OPG expression was increased. This study examined the regulatory mechanism of periostin in bone metabolism, which will provide a future reference for the treatment of primary teeth abnormal replacement caused by inflammation.