AIM: Osteoblasts are key functional cells in the process of bone metabolic balance. Phytoestrogens have an important influence on the proliferation and differentiation of osteoblasts. Puerarin, a plant estrogen, has a...AIM: Osteoblasts are key functional cells in the process of bone metabolic balance. Phytoestrogens have an important influence on the proliferation and differentiation of osteoblasts. Puerarin, a plant estrogen, has a wide range concentration in vitro on the function of osteoblasts. The current study investigates the effect of the phytoestrogen puerarin on the proliferation, differentiation, and mineralization of osteoblasts in vitro. METHODS: The calvaria bone of eight-ten Wistar rats which were born within 24 h were obtained in aseptic condition. After enzyme digestion, isolation, purified osteoblasts of rats were cultured for further study. The cells of the first to third generation were divided into a control group and a puerarin-treated group with 10-3–10-10 mol·L-1 puerarin. The cells were exposed to the medium containing a low level of carbohydrates, 10%(V/V) FBS for 24 h. After 1 to 4 days of culture, the OD values on the proliferation of osteoblasts in each group were determined by microplate reader. The cells were cultured in the medium containing 50 μg·mL-1 vitamin C, 10-2 mol·L-1 sodium glycerophosphate, 10% FBS and the medium was changed every 3 to 4 days. After 2 to 8 days of culture, expression of alkaline phosphatase were tested and compared by microplate reader. The mineral nodes of osteoblasts were dyed using alizarin red or improved Von Kossa way after four weeks. RESULTS: Compared with those in the 10-5–10-9 mol·L-1 puerarin, the proliferation of osteoblasts, the expression of alkaline phosphatase, and the number of mineral nodes of osteoblasts were significantly decreased in the control group. The increase was the fastest in the third day, while on the fourth day it was decreased, and arrived at statistical significance compared with the alkaline phosphatase activities and control group. The 10-6 mol·L-1 group was the most distinct, and formed the most mineralized nodule. Compared with the 10-3 mol·L-1 puerarin group, those changes were markedly increased in the control group. CONC展开更多
基金supported by the Program for New Century Excellent Talents in University(No.NCET-13-0954)the Changjiang Scholars and Innovative Research Team in University(No.IRT1174),the National Natural Science Foundation of China(Nos.J1210063,81001622,81073029)+3 种基金the project"As a Major New Drug to Create a Major National Science and Technology Special"(Nos.2011ZX09401-308-034,2014ZX09304-307-02)the China Postdoctoral Science Foundation(No.2012M521831),the Key Program for the International S&T Cooperation Projects of Shaanxi Province(No.2013KW31-01)the Natural Science Foundation of Shaanxi Provincial Education Department(No.2013JK0811)the Administration of Traditional Chinese Medicine of Shaanxi(No.13-ZY006)
文摘AIM: Osteoblasts are key functional cells in the process of bone metabolic balance. Phytoestrogens have an important influence on the proliferation and differentiation of osteoblasts. Puerarin, a plant estrogen, has a wide range concentration in vitro on the function of osteoblasts. The current study investigates the effect of the phytoestrogen puerarin on the proliferation, differentiation, and mineralization of osteoblasts in vitro. METHODS: The calvaria bone of eight-ten Wistar rats which were born within 24 h were obtained in aseptic condition. After enzyme digestion, isolation, purified osteoblasts of rats were cultured for further study. The cells of the first to third generation were divided into a control group and a puerarin-treated group with 10-3–10-10 mol·L-1 puerarin. The cells were exposed to the medium containing a low level of carbohydrates, 10%(V/V) FBS for 24 h. After 1 to 4 days of culture, the OD values on the proliferation of osteoblasts in each group were determined by microplate reader. The cells were cultured in the medium containing 50 μg·mL-1 vitamin C, 10-2 mol·L-1 sodium glycerophosphate, 10% FBS and the medium was changed every 3 to 4 days. After 2 to 8 days of culture, expression of alkaline phosphatase were tested and compared by microplate reader. The mineral nodes of osteoblasts were dyed using alizarin red or improved Von Kossa way after four weeks. RESULTS: Compared with those in the 10-5–10-9 mol·L-1 puerarin, the proliferation of osteoblasts, the expression of alkaline phosphatase, and the number of mineral nodes of osteoblasts were significantly decreased in the control group. The increase was the fastest in the third day, while on the fourth day it was decreased, and arrived at statistical significance compared with the alkaline phosphatase activities and control group. The 10-6 mol·L-1 group was the most distinct, and formed the most mineralized nodule. Compared with the 10-3 mol·L-1 puerarin group, those changes were markedly increased in the control group. CONC
