Effect of puerarin on proliferation ability of osteoblast MC3T3-E1 cells

FENG Qian; YU PENG; CHENG Songyi; ZENG Xiangwei; YANG Rui; YANG Jing; ZHAO Fengming; ZHU Jiapeng; ZHAN Xiuqin

doi:10.3969/j.issn.1002-266X.2018.16.003

Abstract：Objective To investigate the effect of puerarin on the proliferation ability of osteoblast MC 3T3-E1 and its possible mechanism.Methods MC3T3-E1 cells in the logarithmic growth were selected and treated with different concen-trations of puerarin at different time.CCK-8 was used to detect the effect on cell proliferation.The results showed that the cell proliferation ability was the highest when the cells were treated with 0.1 μmol/L puerarin for 48 h.The Mc3t3-e1 cells were randomly divided into the blank control group, the puerarin group, and the estrogen group, respectively, and were added with the complete medium of α-MEM,0.1 μmol/L puerarin,and 0.01 μmol/L estrogen.After 48 h,we observed cell autophagy under electron microscope.Western blotting was used to detect phosphate-activated mitogen protein kinase (p-AMPK),microtubule associated protein 1 light chain 3(LC3)-Ⅱ/LC3Ⅰexpression.The effects of interfering AMPK on cell autophagy and LC3-Ⅱ/LC3-Ⅰexpression of MC3T3 E1 treated with puerarin.Results The cell organelles,nuclei and chromosome were normal in the blank control group.The cytoplasm of puerarin and oestrogens showed a large number of annular structures,and the substance had bilayer membrane structure specific to autophagy.The expression of p-AMPK and LC3-Ⅱ/LC3-Ⅰin the puerarin group and estrogen group was significantly higher than that of blank control group,and the estrogen group was significantly higher than the puerarin group(all P<0.01).Interfering AMPK expression inhibited puera-rin-mediated MC3T3-E1 cell autophagy,and decreased the LC3-Ⅱ/LC3-Ⅰexpression(all P<0.01).Conclusion Puerarin can enhance the proliferation of MC3T3-E1 cells and the activation of AMPK/LC3 signaling pathway-mediated autophagy is one of the possible mechanisms.