Objective: To observe the effects of Total Flavonoids in Drynaria fortunei (TFDF) on osteoblasts differentiation activity after treatment by high glucose and observe the effects on p38MAPK and ERK1/2 signaling protein in osteoblasts.
Methods: Primary osteoblasts of newborn SD rats was extracted and cultured and its biological characteristics was observed. MTT method was used to observe osteoblasts' cytotoxicity,and to choose a suitable concentration of TFDF in the culture medium. pNPP,ELISA,Alizarin dyeing were used to test ALP,Type I collagen,osteocalcin and mineralization of osteoblasts after treatment by different concentration of glucose respectively and after treatment by TFDF and high glucose. Western-blot was used to detect p38MAPK and ERK1/2 protein phosphorylation after treatment by TFDF and high glucose.
Results: Primary osteoblasts of newborn SD rats could be used well in this experiment. According to the toxicity of TFDF on OB, 25, 50, 100 mg/L of TFDF were selected for the experimental concentration gradient. ALP, Type I collagen,osteocalcin and mineralization of osteoblasts after treatment with glucose (25, 50 mmol/L) were less than those of control group respectively. TFDF could increase ALP, Type I collagen, osteocalcin activity and mineralization of osteoblasts in a dose-dependent manner after treatment by high glucose (25 mmol/L). TFDF(50 mg/L) could increase protein phosphorylation of p38MAPK and ERK1/2 of osteoblasts after treatment by high glucose (25 mmol/L).
Conclusion: High glucose can decrease differentiation and mineralization of osteoblasts. TFDF can increase differentiation and mineralization of osteoblasts in a dose-dependent manner after treatment by high glucose. The role of TFDF in the promotion of osteoblasts differentiation is related to protein phosphorylation of p38MAPK and ERK1/2.