Limits...
Mangiferin reduces the inhibition of chondrogenic differentiation by IL-1β in mesenchymal stem cells from subchondral bone and targets multiple aspects of the Smad and SOX9 pathways.

Huh JE, Koh PS, Seo BK, Park YC, Baek YH, Lee JD, Park DS - Int J Mol Sci (2014)

Bottom Line: Mangiferin is a natural immunomodulator found in plants including mango trees.In IL-1β-stimulated MSCs, mangiferin significantly reversed the production of TGF-β, BMP-2, BMP-4, SOX9, Col2α1, cartilage link protein, and aggrecan, as well as matrix metalloproteinase (MMP)-1, MMP-13, and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS5).In conclusion, mangiferin exhibits both chondrogenic and chondroprotective effects on damaged MSCs and mediates these effects by targeting multiple aspects of the Smad and SOX9 signaling pathways.

View Article: PubMed Central - PubMed

Affiliation: East-West Bone & Joint Research Institute, Kyung Hee University, 149, Sangil-dong, Gangdong-gu, Seoul 134-727, Korea. jehuh2551@hanmail.net.

ABSTRACT
Mangiferin is a natural immunomodulator found in plants including mango trees. The effects of mangiferin on chondrogenesis and cartilage repair have not yet been reported. This study was designed to determine the effect of mangiferin on chondrogenic differentiation in IL-1β-stimulated mesenchymal stem cells (MSCs) from subchondral bone and to explore the mechanisms underlying these effects. MSCs were isolated from the subchondral bone of rabbit and treated with mangiferin alone and/or interleukin-1β (IL-1β). Mangiferin induced chondrogenic differentiation in MSCs by upregulating transforming growth factor (TGF)-β, bone morphogenetic protein (BMP)-2, and BMP-4 and several key markers of chondrogenesis, including sex-determining region Y-box (SRY-box) containing gene 9 (SOX9), type 2α1 collagen (Col2α1), cartilage link protein, and aggrecan. In IL-1β-stimulated MSCs, mangiferin significantly reversed the production of TGF-β, BMP-2, BMP-4, SOX9, Col2α1, cartilage link protein, and aggrecan, as well as matrix metalloproteinase (MMP)-1, MMP-13, and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS5). Mangiferin upregulated the phosphorylation of Smad 2, Smad 3, Smad 1/5/8, and SOX9 in IL-1β-stimulated MSCs. In the presence of mangiferin, SOX9 siRNA suppressed the activation of Smad 2, Smad 3, Smad 1/5/8, aggrecan, and Col2α1 expression. In conclusion, mangiferin exhibits both chondrogenic and chondroprotective effects on damaged MSCs and mediates these effects by targeting multiple aspects of the Smad and SOX9 signaling pathways.

Show MeSH

Related in: MedlinePlus

Effects of mangiferin on the phosphorylation of the Smad and SOX9 signaling pathways in IL-1β-stimulated mesenchymal stem cells (MSCs). (A) Mangiferin upregulates the phosphorylation of Smad 2, Smad 3, Smad 1/5/8, and the SOX9 signaling pathways. MSCs were prestimulated with IL-1β (5 ng/mL) for 1 h and then treated with the indicated dose of mangiferin for 72 h. Whole cell lysates were subjected to Western blotting with the indicated antibodies; (B,C) SOX9 deficiency inhibits the phosphorylation of Smad 2, Smad 3, Smad 1/5/8, aggrecan, and Col2α1. β-actin served as an internal control. MSCs were transfected with control siRNA or SOX9 siRNA and then treated with or without mangiferin for 72 h. Whole cell lysates were subjected to Western blotting with the indicated antibodies. siRNA targeting SOX9 was transfected into two different experiments.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4200868&req=5

ijms-15-16025-f005: Effects of mangiferin on the phosphorylation of the Smad and SOX9 signaling pathways in IL-1β-stimulated mesenchymal stem cells (MSCs). (A) Mangiferin upregulates the phosphorylation of Smad 2, Smad 3, Smad 1/5/8, and the SOX9 signaling pathways. MSCs were prestimulated with IL-1β (5 ng/mL) for 1 h and then treated with the indicated dose of mangiferin for 72 h. Whole cell lysates were subjected to Western blotting with the indicated antibodies; (B,C) SOX9 deficiency inhibits the phosphorylation of Smad 2, Smad 3, Smad 1/5/8, aggrecan, and Col2α1. β-actin served as an internal control. MSCs were transfected with control siRNA or SOX9 siRNA and then treated with or without mangiferin for 72 h. Whole cell lysates were subjected to Western blotting with the indicated antibodies. siRNA targeting SOX9 was transfected into two different experiments.

Mentions: We hypothesized that the effect of mangiferin on TGF-β and BMP expression and induction of chondrogenesis is likely to be related to the regulation of Smad signaling. To test this, we determined the involvement of mangiferin in the Smad and SOX9 signaling pathways. Western blot analysis of IL-1β-stimulated MSCs revealed that the chondrogenic differentiation process was disturbed due to the downregulation of phosphorylation of Smad 2, Smad 3, Smad 1/5/8, and SOX9. Mangiferin treatment increased the phosphorylation of Smad 2, Smad 3, and Smad 1/5/8 in IL-1β-stimulated MSCs (Figure 5A). Smads are associated with the TGF-β and BMP signaling pathways, which, in turn, are involved with MSCs differentiation [38,39,40,41]. Smad 2 and Smad 3 are known to mediate transcription processes in the TGF-β signaling pathway, and Smad 1/5/8 is known to mediate transcription in the BMP signaling pathway, leading to regulation of the transcription factor SOX9 [38,39,40]. SOX9 plays a key role in chondrogenesis and controls the expression of aggrecan and type II collagen [42,43]. To determine the involvement of SOX9 in the expression of aggrecan and Col2α1, IL-1β-stimulated MSCs were transfected with control siRNA or SOX9 siRNA and then treated with mangiferin. In the presence of mangiferin and SOX9 siRNA, phosphorylation of Smad 2, Smad 3, Smad 1/5/8, aggrecan, and Col2α1 was suppressed in IL-1β-stimulated MSCs (Figure 5B,C). This suggests that SOX9 is an important pathway for the chondrogenic differentiation process, and that mangiferin may act on SOX9 to normalize chondrogenic differentiation and chondrogenesis. Thus, the present results indicate that mangiferin affects chondrogenesis by stimulating the SOX9 and by inducing the phosphorylation of Smad 1/5/8 and Smad 2/3 signaling pathways.


Mangiferin reduces the inhibition of chondrogenic differentiation by IL-1β in mesenchymal stem cells from subchondral bone and targets multiple aspects of the Smad and SOX9 pathways.

Huh JE, Koh PS, Seo BK, Park YC, Baek YH, Lee JD, Park DS - Int J Mol Sci (2014)

Effects of mangiferin on the phosphorylation of the Smad and SOX9 signaling pathways in IL-1β-stimulated mesenchymal stem cells (MSCs). (A) Mangiferin upregulates the phosphorylation of Smad 2, Smad 3, Smad 1/5/8, and the SOX9 signaling pathways. MSCs were prestimulated with IL-1β (5 ng/mL) for 1 h and then treated with the indicated dose of mangiferin for 72 h. Whole cell lysates were subjected to Western blotting with the indicated antibodies; (B,C) SOX9 deficiency inhibits the phosphorylation of Smad 2, Smad 3, Smad 1/5/8, aggrecan, and Col2α1. β-actin served as an internal control. MSCs were transfected with control siRNA or SOX9 siRNA and then treated with or without mangiferin for 72 h. Whole cell lysates were subjected to Western blotting with the indicated antibodies. siRNA targeting SOX9 was transfected into two different experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4200868&req=5

ijms-15-16025-f005: Effects of mangiferin on the phosphorylation of the Smad and SOX9 signaling pathways in IL-1β-stimulated mesenchymal stem cells (MSCs). (A) Mangiferin upregulates the phosphorylation of Smad 2, Smad 3, Smad 1/5/8, and the SOX9 signaling pathways. MSCs were prestimulated with IL-1β (5 ng/mL) for 1 h and then treated with the indicated dose of mangiferin for 72 h. Whole cell lysates were subjected to Western blotting with the indicated antibodies; (B,C) SOX9 deficiency inhibits the phosphorylation of Smad 2, Smad 3, Smad 1/5/8, aggrecan, and Col2α1. β-actin served as an internal control. MSCs were transfected with control siRNA or SOX9 siRNA and then treated with or without mangiferin for 72 h. Whole cell lysates were subjected to Western blotting with the indicated antibodies. siRNA targeting SOX9 was transfected into two different experiments.
Mentions: We hypothesized that the effect of mangiferin on TGF-β and BMP expression and induction of chondrogenesis is likely to be related to the regulation of Smad signaling. To test this, we determined the involvement of mangiferin in the Smad and SOX9 signaling pathways. Western blot analysis of IL-1β-stimulated MSCs revealed that the chondrogenic differentiation process was disturbed due to the downregulation of phosphorylation of Smad 2, Smad 3, Smad 1/5/8, and SOX9. Mangiferin treatment increased the phosphorylation of Smad 2, Smad 3, and Smad 1/5/8 in IL-1β-stimulated MSCs (Figure 5A). Smads are associated with the TGF-β and BMP signaling pathways, which, in turn, are involved with MSCs differentiation [38,39,40,41]. Smad 2 and Smad 3 are known to mediate transcription processes in the TGF-β signaling pathway, and Smad 1/5/8 is known to mediate transcription in the BMP signaling pathway, leading to regulation of the transcription factor SOX9 [38,39,40]. SOX9 plays a key role in chondrogenesis and controls the expression of aggrecan and type II collagen [42,43]. To determine the involvement of SOX9 in the expression of aggrecan and Col2α1, IL-1β-stimulated MSCs were transfected with control siRNA or SOX9 siRNA and then treated with mangiferin. In the presence of mangiferin and SOX9 siRNA, phosphorylation of Smad 2, Smad 3, Smad 1/5/8, aggrecan, and Col2α1 was suppressed in IL-1β-stimulated MSCs (Figure 5B,C). This suggests that SOX9 is an important pathway for the chondrogenic differentiation process, and that mangiferin may act on SOX9 to normalize chondrogenic differentiation and chondrogenesis. Thus, the present results indicate that mangiferin affects chondrogenesis by stimulating the SOX9 and by inducing the phosphorylation of Smad 1/5/8 and Smad 2/3 signaling pathways.

Bottom Line: Mangiferin is a natural immunomodulator found in plants including mango trees.In IL-1β-stimulated MSCs, mangiferin significantly reversed the production of TGF-β, BMP-2, BMP-4, SOX9, Col2α1, cartilage link protein, and aggrecan, as well as matrix metalloproteinase (MMP)-1, MMP-13, and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS5).In conclusion, mangiferin exhibits both chondrogenic and chondroprotective effects on damaged MSCs and mediates these effects by targeting multiple aspects of the Smad and SOX9 signaling pathways.

View Article: PubMed Central - PubMed

Affiliation: East-West Bone & Joint Research Institute, Kyung Hee University, 149, Sangil-dong, Gangdong-gu, Seoul 134-727, Korea. jehuh2551@hanmail.net.

ABSTRACT
Mangiferin is a natural immunomodulator found in plants including mango trees. The effects of mangiferin on chondrogenesis and cartilage repair have not yet been reported. This study was designed to determine the effect of mangiferin on chondrogenic differentiation in IL-1β-stimulated mesenchymal stem cells (MSCs) from subchondral bone and to explore the mechanisms underlying these effects. MSCs were isolated from the subchondral bone of rabbit and treated with mangiferin alone and/or interleukin-1β (IL-1β). Mangiferin induced chondrogenic differentiation in MSCs by upregulating transforming growth factor (TGF)-β, bone morphogenetic protein (BMP)-2, and BMP-4 and several key markers of chondrogenesis, including sex-determining region Y-box (SRY-box) containing gene 9 (SOX9), type 2α1 collagen (Col2α1), cartilage link protein, and aggrecan. In IL-1β-stimulated MSCs, mangiferin significantly reversed the production of TGF-β, BMP-2, BMP-4, SOX9, Col2α1, cartilage link protein, and aggrecan, as well as matrix metalloproteinase (MMP)-1, MMP-13, and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS5). Mangiferin upregulated the phosphorylation of Smad 2, Smad 3, Smad 1/5/8, and SOX9 in IL-1β-stimulated MSCs. In the presence of mangiferin, SOX9 siRNA suppressed the activation of Smad 2, Smad 3, Smad 1/5/8, aggrecan, and Col2α1 expression. In conclusion, mangiferin exhibits both chondrogenic and chondroprotective effects on damaged MSCs and mediates these effects by targeting multiple aspects of the Smad and SOX9 signaling pathways.

Show MeSH
Related in: MedlinePlus