The experiment study of the effects of icariin on Smad1, 5 in MC3T3-E1 cell in vitro.

Abstract

Objective To explore the effect of icariin on the expression of Smad1, 5 mRNA and protein Smad3, 5 in MC3T3-E1 cells in vitro. Methods According to the stimulus concentrations (0, 30, 40 and 80 ng/ml) of icariin, the MC3T3-E1 cells were divided into four groups. After stimulated by icariin 24, 48 and 72 h, the RT-PCR was used to detect the mRNA expression level about Smad1, 5. Western blot technique was applied to exsplore the Smad1, 5 protein expression. The immunohistochemistry was engaged to comfirm and to localize the expression of Smad1, 5 protein in MC3T3-E1 cells. All data were treated with One-way Anova analysis and Dunnett-t test in SPSS 13.0 software. Results After stimulated by inariin of 48 h and 72 h, both Smad1, 5 mRNA increased continuously by time in 10, 40 and 80 ng/ml, the difference were statistical significance. But in 0 ng/ml group, Smad1, 5 mRNA was no obviously increased. At 72 h, samd1, 5 mRNA were not observed expression in 0 ng/ml groups. Western blot demonstrated that Smad1 and protein under the stimulation of different dose of icariin expressed much more than 0 ng/ml groups at differ-ent times. Western blot shown that Smadl, 5 protein level were higher in 30, 40 and 80 ng/ml, than that in 0 ng/ml group at 24, 48 or 72 h, the difference was statistical significance. Immunohistochemistry dis-played that in 10, 40 and 80 ng/ml groups, the expression both of the Smad1, 5 proteins were strongly pos-tive in cytoplasm and nuclei, but was mildly positve in 0 ng/ml group. Conclusion Icariin is able to up-regulate the expression level of Smad1, 5 mRNA and protein in vitro. Key words: Icariin; Osteoblasts; Cell culture techniques