| Abstract: Objective To observe the effect of hirudin on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Methods BMSCs cells were divided into normal cultured control group, osteogenic induced group, and different hirudin groups (1, 10, 20 ATU/mL). MTT method was used to detect cell proliferation and screen the optimal concentration of hirudin. Cell apoptosis was measured using flow cytometry method. mRNA and protein expressions of Runx2, Osterix, and COL1A1 were measured using RT-PCR and Western blotting, respectively. Alkaline phosphatase was detected with BCIP/NBT staining. Alizarin red staining was used to detect mineralized nodules. mRNA and protein expressions of VEGF, Notch1, Jagged1, and CBF1 were measured. Results BMSCs cell proliferation increased significantly after osteogenic induction. Hirudin at medium/high concentration elevated cell proliferation of osteogenic induced BMSCs (P<0.05), and 20 ATU/mL was selected as the optimal concentration of hirudin. Hirudin decreased cell apoptosis, elevated mRNA and protein expression level of Runx2, Osterix, and COL1A1, increased alkaline phosphatase levels, promoted the formation of mineralized nodules, and increased expressions of VEGF, Notch1, Jagged1, and CBF1 in osteogenic induced BMSCs (P<0.05). Conclusion Hirudin may promote osteogenic differentiation of bone marrow mesenchymal stem cells by up-regulating VEGF/Notch1 signaling pathway. |