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Hepcidin promotes osteogenic differentiation through the bone morphogenetic protein 2/small mothers against decapentaplegic and mitogen-activated protein kinase/P38 signaling pathways in mesenchymal stem cells.

Lu H, Lian L, Shi D, Zhao H, Dai Y - Mol Med Rep (2014)

Bottom Line: The present study demonstrated that hepcidin enhanced osteoblastic differentiation and mineralization, which was manifested by an upregulation in the differentiation markers alkaline phosphatase and osteogenic genes.In addition, the p38 mitogen-activated protein kinase may be an upstream kinase for osteoblastic differentiation.Thus, hepcidin may be important in the osteogenic differentiation of MSCs and may be considered as a target in the development of therapies for pathological bone loss.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedics, Third Affiliated Hospital of Sun Yat‑sen University, Guangzhou, Guangdong 510630, P.R. China.

ABSTRACT
The ability of mesenchymal stem cells (MSCs) to differentiate into osteogenic lineages requires management for their future use in treating bone destruction and osteoporosis. Hepcidin is closely associated with bone metabolism, however, it remains to be elucidated whether hepcidin affects osteogenic differentiation in MSCs. The present study demonstrated that hepcidin enhanced osteoblastic differentiation and mineralization, which was manifested by an upregulation in the differentiation markers alkaline phosphatase and osteogenic genes. Furthermore, the expression levels of bone morphogenetic proteins and small mothers against decapentaplegic homologs were concomitantly increased following hepcidin treatment. In addition, the p38 mitogen-activated protein kinase may be an upstream kinase for osteoblastic differentiation. Thus, hepcidin may be important in the osteogenic differentiation of MSCs and may be considered as a target in the development of therapies for pathological bone loss.

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Related in: MedlinePlus

Effect of hepcidin on the BMP2/Smads pathway in MSCs. (A) MSCs were cultured in osteogenic medium with or without 0.2 mmol/l hepcidin for 24, 48 and 72 h and the expression level of BMP2 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control (**P<0.01, vs. control; #P<0.01, vs. control). (B) Cells were cultured, as described above, and the expression level of Smad1 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control. (**P<0.01, vs. control; #P<0.01, vs. control). (C) Cells were cultured, as described above, and the expression level of Smad5 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control (**P<0.01, vs. control 24 h; #P<0.01, vs. control). (D) Cells were cultured, as described above, and the expression level of Smad8 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control (**P<0.01, vs. control; #P<0.01, vs. control). MSC, mesenchymal stem cell; BMP2, bone morphogenetic protein 2; Smads; small mothers against decapentaplegic; RT-qPCR, reverse transcription polymerase chain reaction; SEM, standard error of the mean.
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f2-mmr-11-01-0143: Effect of hepcidin on the BMP2/Smads pathway in MSCs. (A) MSCs were cultured in osteogenic medium with or without 0.2 mmol/l hepcidin for 24, 48 and 72 h and the expression level of BMP2 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control (**P<0.01, vs. control; #P<0.01, vs. control). (B) Cells were cultured, as described above, and the expression level of Smad1 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control. (**P<0.01, vs. control; #P<0.01, vs. control). (C) Cells were cultured, as described above, and the expression level of Smad5 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control (**P<0.01, vs. control 24 h; #P<0.01, vs. control). (D) Cells were cultured, as described above, and the expression level of Smad8 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control (**P<0.01, vs. control; #P<0.01, vs. control). MSC, mesenchymal stem cell; BMP2, bone morphogenetic protein 2; Smads; small mothers against decapentaplegic; RT-qPCR, reverse transcription polymerase chain reaction; SEM, standard error of the mean.

Mentions: Previous studies have demonstrated that BMP/Smad signaling pathway usually mediates osteoblastic differentiation and maturation (21). Therefore, the present study examined whether the treatment of MSCs with hepcidin affected the BMP/Smad pathways in this process. The MSCs were cultured in osteogenic medium with or without 0.2 mmol/l hepcidin for 24, 48 and 72 hours, The expression levels of BMP2 mRNA, Smad1 mRNA, Smad5 mRNA and Smad8 mRNA were detected using RT-qPCR. As shown in Fig. 2, the BMP2, Smad1, Smad5 and Smad8 mRNAs were significantly upregulated by hepcidin treatment compared with the control groups, following culture for 24, 48 and 72 h. Overall, these results suggested that hepcidin activated the BMP2/Smad pathway.


Hepcidin promotes osteogenic differentiation through the bone morphogenetic protein 2/small mothers against decapentaplegic and mitogen-activated protein kinase/P38 signaling pathways in mesenchymal stem cells.

Lu H, Lian L, Shi D, Zhao H, Dai Y - Mol Med Rep (2014)

Effect of hepcidin on the BMP2/Smads pathway in MSCs. (A) MSCs were cultured in osteogenic medium with or without 0.2 mmol/l hepcidin for 24, 48 and 72 h and the expression level of BMP2 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control (**P<0.01, vs. control; #P<0.01, vs. control). (B) Cells were cultured, as described above, and the expression level of Smad1 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control. (**P<0.01, vs. control; #P<0.01, vs. control). (C) Cells were cultured, as described above, and the expression level of Smad5 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control (**P<0.01, vs. control 24 h; #P<0.01, vs. control). (D) Cells were cultured, as described above, and the expression level of Smad8 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control (**P<0.01, vs. control; #P<0.01, vs. control). MSC, mesenchymal stem cell; BMP2, bone morphogenetic protein 2; Smads; small mothers against decapentaplegic; RT-qPCR, reverse transcription polymerase chain reaction; SEM, standard error of the mean.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4237087&req=5

f2-mmr-11-01-0143: Effect of hepcidin on the BMP2/Smads pathway in MSCs. (A) MSCs were cultured in osteogenic medium with or without 0.2 mmol/l hepcidin for 24, 48 and 72 h and the expression level of BMP2 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control (**P<0.01, vs. control; #P<0.01, vs. control). (B) Cells were cultured, as described above, and the expression level of Smad1 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control. (**P<0.01, vs. control; #P<0.01, vs. control). (C) Cells were cultured, as described above, and the expression level of Smad5 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control (**P<0.01, vs. control 24 h; #P<0.01, vs. control). (D) Cells were cultured, as described above, and the expression level of Smad8 mRNA was detected using RT-qPCR. The data are expressed as the means ± SEM (n=3). GAPDH was used as a loading control (**P<0.01, vs. control; #P<0.01, vs. control). MSC, mesenchymal stem cell; BMP2, bone morphogenetic protein 2; Smads; small mothers against decapentaplegic; RT-qPCR, reverse transcription polymerase chain reaction; SEM, standard error of the mean.
Mentions: Previous studies have demonstrated that BMP/Smad signaling pathway usually mediates osteoblastic differentiation and maturation (21). Therefore, the present study examined whether the treatment of MSCs with hepcidin affected the BMP/Smad pathways in this process. The MSCs were cultured in osteogenic medium with or without 0.2 mmol/l hepcidin for 24, 48 and 72 hours, The expression levels of BMP2 mRNA, Smad1 mRNA, Smad5 mRNA and Smad8 mRNA were detected using RT-qPCR. As shown in Fig. 2, the BMP2, Smad1, Smad5 and Smad8 mRNAs were significantly upregulated by hepcidin treatment compared with the control groups, following culture for 24, 48 and 72 h. Overall, these results suggested that hepcidin activated the BMP2/Smad pathway.

Bottom Line: The present study demonstrated that hepcidin enhanced osteoblastic differentiation and mineralization, which was manifested by an upregulation in the differentiation markers alkaline phosphatase and osteogenic genes.In addition, the p38 mitogen-activated protein kinase may be an upstream kinase for osteoblastic differentiation.Thus, hepcidin may be important in the osteogenic differentiation of MSCs and may be considered as a target in the development of therapies for pathological bone loss.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedics, Third Affiliated Hospital of Sun Yat‑sen University, Guangzhou, Guangdong 510630, P.R. China.

ABSTRACT
The ability of mesenchymal stem cells (MSCs) to differentiate into osteogenic lineages requires management for their future use in treating bone destruction and osteoporosis. Hepcidin is closely associated with bone metabolism, however, it remains to be elucidated whether hepcidin affects osteogenic differentiation in MSCs. The present study demonstrated that hepcidin enhanced osteoblastic differentiation and mineralization, which was manifested by an upregulation in the differentiation markers alkaline phosphatase and osteogenic genes. Furthermore, the expression levels of bone morphogenetic proteins and small mothers against decapentaplegic homologs were concomitantly increased following hepcidin treatment. In addition, the p38 mitogen-activated protein kinase may be an upstream kinase for osteoblastic differentiation. Thus, hepcidin may be important in the osteogenic differentiation of MSCs and may be considered as a target in the development of therapies for pathological bone loss.

Show MeSH
Related in: MedlinePlus