关键词: 抑瘤素M, 脐带间充质干细胞, 成骨, 细胞增殖, 细胞分化

Abstract: Objective: To investigate the effect of oncostatin M (OSM) on the proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells(hUCMSCs), and to elucidate that OSM is an effective osteogenic induction active factor. Methods: The hUCMSCs were cultivated in vitro. The surface antigens of hUCMSCs at the third generation were detected by flow cytometry. CCK-8 assay was used to determine the proliferation activities of hUCMSCs after treated by 0.1,1.0 and 10.0 μg·L^-1 recombined human oncostatin M(rhOSM).The hUCMSCs were divided into control group,classical osteogenic induction group,and different doses (0.1,1.0 and 10.0 μg·L^-1) of rhOSM osteogenic induction groups. Alkaline phosphatase(ALP) was stained by BCIP/NBT and the ALP activities in the cells were determined quantitatively on the 4th,7th and 14th days of osteogenic induction; Alizarin Red S staining (ARS) was performed to assess the formation of calcium nodules and the cell mineralization activities were semi-quantitatively analyzed. RT-qPCR was carried out to measure the expression levels of Runt-related transcription factor 2(Runx2) mRNA and OCN mRNA in the hUCMSCs. Results: The hUCMSCs were consistent with the identification criteria of mesenchymal stem cells(MSCs).After treatment with rhOSM for 72 h, compared with control group, the proliferation activity of hUCMSCs in 10.0 μg·L^-1 rhOSM group was decreased (P<0.05); after treated for 96 h, the proliferation activities in different doses of rhOSM groups were significantly lower than that in control group (P<0.05); there were significant differences between different doses of rhOSM groups(P<0.01).On the 4th, 7th and 14th days,compared with classical osteogenic induction group,the ALP activities and the expression levels of Runx2 mRNA in the hUCMSCs in different doses of rhOSM groups were increased(P<0.01).Compared with 0.1 μg·L^-1 rhOSM group,the ALP activities and the expression levels of Runx2 mRNA in the hUCMSCs in 1.0 and 10.0 μg·L^-1 rhOSM groups were significantly increased(P<0.05). Compared with the 7th day of induction,the ALP activities and the expression levels of Runx2 mRNA in different doses of rhOSM groups were slightly decreased on the 14th day of induction, but there were no significant differences(P<0.05).On the 7th,14th and 21st days,compared with classical osteogenic induction group, the mineralization activities in the hUCMSCs in different doses of rhOSM groups were increased(P<0.01);compared with 0.1 μg·L^-1 rhOSM group,the mineralization activities in the hUCMSCs in 1.0 and 10.0 μg·L^-1 rhOSM groups were significantly increased(P<0.05).Compared with classical osteogenic induction group,the expression levels of OCN mRNA ih the hUCMSCs in different doses of rhOSM groups were significatnly increased(P<0.05);on the 7th,14th and 21st days of induction,compared with 0.1 μg·L^-1 rhOSM grouop,the expression levels of OCN mRNA in the cells in 1.0 and 10.0 μg·L^-1 rhOSM groups were signficantly increased(P<0.05) in a time-dose manner. Conclusion: rhOSM can promote the osteogenic differentiation of hUCMSCs in vitro by increasing the expressions of Runx2 and OCN and increasing the ALP activity and the deposition of calcium salts and is an effective osteogenic induction active factor.

Key words: oncostatin M, umbilical cord mesenchymal stem cells, osteogenesis, cell proliferation, cell differentiation