Insulin as a Potent Stimulator of Akt, ERK and Inhibin-βE Signaling in Osteoblast-Like UMR-106 Cells

Insulin is a peptide hormone of the endocrine pancreas and exerts a wide variety of physiological actions in insulin sensitive tissues, such as regulation of glucose homeostasis, cell growth, differentiation, learning and memory. However, the role of insulin in osteoblast cells remains to be fully characterized. In this study, we demonstrated that the insulin (100 nM) has the ability to stimulate the phosphorylation of protein kinase B (Akt/PKB) and extracellular signal-regulated kinase (ERK) and the levels of inhibin-βE in the osteoblast-like UMR-106 cells. This insulin-stimulated activities were abolished by the PI3K and MEK1 inhibitors LY294002 and PD98059, respectively. This is the first report proving that insulin is a potential candidate that enables the actions of inhibin-βE subunit of the TGF-β family. The current investigation provides a foundation for the realization of insulin as a potential stimulator in survival signaling pathways in osteoblast-like UMR-106 cells.

Chemical constituents isolated from fruits of Cnidium monnieri and their effects on proliferation of UMR106 cells / 中草药

Objective: To study the chemical constituents from the fruits of Cnidium monnieri and their effects on proliferation of UMR106 cells. Methods: The constituents were separated by column chromatography, and their structures were elucidated by spectroscopic data analyses. The proliferation of all isolated compounds on osteoblast-like UMR106 cells was determined. Results: Nine compounds were isolated and identified as 5,7-dihydroxy-6,8-dimethoxy-2-methyl-4H-chromen-4-one (1), 5-hydroxy-2- hydroxymethyl-4H-chromen-4-one (2), (+)-marmesin (3), bergaptol (4), isobergapten (5), 7-hydroxy-8-(2',3'-dihydroxy-3'-methyl-butyl)-coumarin (6), murraol (7), coniferyl aldehyde (8), and m-hydroxybenzoic acid (9). Compounds 1, 3, and 7 showed the significant proliferative activities on UMR106 cells lines at the concentration of 1 × 10-10 mol/L and the proliferative ratios were 31.55%, 32.39% and 30.87%. Conclusion: Compounds 1-6 are isolated from the specie of Cnidium Cusson for the first time. Compounds 1, 3, and 7 increase UMR106 cells proliferation to some extent.

Beneficial effects of low glycosides fraction from Epimedii Herba on UMR-106 cells and zebrafish with osteoporosis / 中草药

Objective: To assess the potential effect of the low glycosides fraction from Epimedii Herba (LGFEH) on UMR-106 cells and zebrafish model with osteoporosis induced by Prednisolone. Methods: The MTT assay was used to detect the effect of LGFEH on the proliferation of UMR-106 cells, and at the same time, the influence of LGFEH to the UMR-106 cells differentiation was observed through testing the ALP activity of UMR-106 cells using biochemical assays. Zebrafish larvae at 3 d post fertilization were divided into blank control group, 0.5% DMSO group, Prednisolone group, etidronate disodium group, and LGFEH (1, 2, 4, 8, and 16 μg/mL) groups. All groups were incubated in 24-well plates for 9 d until execution, and then zebrafish skeleton was anesthetized and fixed for staining with alizarin red. Quantitative analysis of the stained area was performed by microscopic inspection and digital imaging methods to reflect the amount of bone mineralization. Results: The LGFEH significantly increased both proliferation and ALP activities of UMR-106 cells. Furthermore, compared with model group, head skeleton mineral area and IOD of the LGFEH groups were significantly increased. Conclusion: Our results indicate that the LGFEH might be beneficial for treating osteoporosis.

Chomones from fruit of cnidium monnieri and their effects on proliferation of UMR106 cells / 中草药

Objective: To study the chomenes from the fruit of Cnidium monnieri and their effects on proliferation of UMR106 cells. Methods: The constituents were separated by column chromatography, and their structures were elucidated by spectroscopic data analyses. The effects of all isolated compounds on proliferation of osteoblast-like UMR106 cells were determined. Results: Ten compounds were isolated and identified as cnidimoside A (1), cnidimol B (2), peucenin (3), 5,7-dihydroxychromone (4), 5-O-methylvisamminol (5), 4'-O-β-D-glucosyl-5-O-methylvisamminol (6), hamaudol (7), 2,5-dimethyl-7-hydroxychromone (8), cimifugin (9), and 5-hydroxy-chromone-7-O-β-D-glucoside (10). Compounds 1,5,9, and 10 showed the significant proliferative activities against UMR106 cells lines at the concentration of 0.10 nmol/L and the proliferative ratios were 30.23%, 31.56%, 35.29%, and 33.36%. Conclusion: Compounds 3-10 are isolated from species of genus Cnidium Cuss. for the first time. Compounds 1,5,9, and 10 (0.10 nmol/L) could increase the proliferation of UMR106 cells to some extent.

Fibroblast growth factor-induced Thymidylate Synthase activity and expression in the serum-starved UMR 106-01 osteoblast cells / 대한정형외과연구학회지

PURPOSE: In the present study, the effects of bFGF on the early responses of proliferation of UMR 106-01 osteoblast cells during cell cycle reentry from the latent(G0/G1) to the proliferative periods(S/M) were investigated. MATERIALS AND METHODS: The synchronized cell culture method using the serum starvation was utilized. After the addition of bFGF, the time courses of protein synthesis, DNA synthesis, thymidylate synthase(TS) activity, TS mRNA level and expression of c-fos were determined. RESULTS: 87% UMR 106-01 cells were synchronized to G0/G1 by serum starvation for seven days in the medium containing 0.1% serum. The protein level began to increase 3 hours after bFGF treatment and reached the maximum at 18 hours. TS activity began to increase 3 hours after the bFGF treatment and reached its peak at 6 hours while its mRNA level, determined by quantitative PCR, reached the maximum at 12 hours. The expression of c-fos protein, determined by western blot analysis and immunocytochemistry, increased 3 hours after bFGF treatment. On the contrary, these prominent changes and responses to bFGF were not observed in the case of using non-synchronized cells cultured in the medium containing 10% serum. CONCLUSION: Based on these data it can be concluded that bFGF-induced DNA synthesis in the early proliferative phase is due to increases in both TS activity and mRNA amount and that the increase in c-fos expression and TS activity occur before the increase in TS mRNA level.

Fibroblast growth factor-induced Thymidylate Synthase activity and expression in the serum-starved UMR 106-01 osteoblast cells / 대한정형외과연구학회지

PURPOSE: In the present study, the effects of bFGF on the early responses of proliferation of UMR 106-01 osteoblast cells during cell cycle reentry from the latent(G0/G1) to the proliferative periods(S/M) were investigated. MATERIALS AND METHODS: The synchronized cell culture method using the serum starvation was utilized. After the addition of bFGF, the time courses of protein synthesis, DNA synthesis, thymidylate synthase(TS) activity, TS mRNA level and expression of c-fos were determined. RESULTS: 87% UMR 106-01 cells were synchronized to G0/G1 by serum starvation for seven days in the medium containing 0.1% serum. The protein level began to increase 3 hours after bFGF treatment and reached the maximum at 18 hours. TS activity began to increase 3 hours after the bFGF treatment and reached its peak at 6 hours while its mRNA level, determined by quantitative PCR, reached the maximum at 12 hours. The expression of c-fos protein, determined by western blot analysis and immunocytochemistry, increased 3 hours after bFGF treatment. On the contrary, these prominent changes and responses to bFGF were not observed in the case of using non-synchronized cells cultured in the medium containing 10% serum. CONCLUSION: Based on these data it can be concluded that bFGF-induced DNA synthesis in the early proliferative phase is due to increases in both TS activity and mRNA amount and that the increase in c-fos expression and TS activity occur before the increase in TS mRNA level.