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MCP-1 downregulates MMP-9 export via vesicular redistribution to lysosomes in rat portal fibroblasts.

Hickman DA, Syal G, Fausther M, Lavoie EG, Goree JR, Storrie B, Dranoff JA - Physiol Rep (2014)

Bottom Line: We examined the effect of MCP-1 on release of matrix metalloproteinase-9 (MMP-9) by rat PF.We found that MCP-1 blocks PF release of MMP-9 in a posttranslational fashion.Our data demonstrated that, in the presence of MCP-1, MMP-9-containing vesicles were shunted to a lysosome-like compartment.

View Article: PubMed Central - PubMed

Affiliation: Division of Gastroenterology and Hepatology, Department of Internal Medicine, University of Arkansas for Medical Sciences, Little Rock, Arkansas.

No MeSH data available.


Related in: MedlinePlus

Effect of MCP‐1 on PF supernatant MMP‐9 activity as determined by gelatin zymography. (A) Representative zymogram. Primary rat PF were isolated and either left untreated or were treated overnight with MCP‐1 (0.1–50 ng/mL). Cells were washed, and the cell supernatant was collected after 30 min. As can be seen, the relative activity of MMP‐9 (seen at 92 KDa) decreases after MCP‐1 treatment. In contrast, MMP2 activity (seen at 72 KDa) is unaltered. (B) Quantitation of data. Band intensity was quantitated electronically, and changes in band intensity are shown. MCP‐1 decreased MMP‐9 band intensity to 38–53% relative to untreated cells.
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fig01: Effect of MCP‐1 on PF supernatant MMP‐9 activity as determined by gelatin zymography. (A) Representative zymogram. Primary rat PF were isolated and either left untreated or were treated overnight with MCP‐1 (0.1–50 ng/mL). Cells were washed, and the cell supernatant was collected after 30 min. As can be seen, the relative activity of MMP‐9 (seen at 92 KDa) decreases after MCP‐1 treatment. In contrast, MMP2 activity (seen at 72 KDa) is unaltered. (B) Quantitation of data. Band intensity was quantitated electronically, and changes in band intensity are shown. MCP‐1 decreased MMP‐9 band intensity to 38–53% relative to untreated cells.

Mentions: We investigated the effect of MCP‐1 on MMP‐9 activity via gelatin zymography. As seen in Figure 1, MCP‐1 decreased MMP‐9 activity in PF in a concentration‐sensitive fashion. Note that no change in MMP‐2 activity was observed, consistent with the concept that, in general, MMP‐9 is regulated (Hatfield et al. 2010), whereas MMP2 is constitutive (Gioia et al. 2009). We then examined whether the observed downregulation of MMP‐9 activity was due to downregulation of transcription. As seen in Figure 2A, MCP‐1 induced no change in MMP‐9 mRNA. Finally, we investigated whether MCP‐1 regulated the intracellular protein content of MMP‐9. We found that MMP‐9 protein could be detected in PF only after treatment with MCP‐1 (10 or 100 ng/mL). Taken together, these findings suggested that MCP‐1 altered PF MMP‐9 activity in a fashion independent of transcriptional changes and possibly due to alteration of protein release into the extracellular environment.


MCP-1 downregulates MMP-9 export via vesicular redistribution to lysosomes in rat portal fibroblasts.

Hickman DA, Syal G, Fausther M, Lavoie EG, Goree JR, Storrie B, Dranoff JA - Physiol Rep (2014)

Effect of MCP‐1 on PF supernatant MMP‐9 activity as determined by gelatin zymography. (A) Representative zymogram. Primary rat PF were isolated and either left untreated or were treated overnight with MCP‐1 (0.1–50 ng/mL). Cells were washed, and the cell supernatant was collected after 30 min. As can be seen, the relative activity of MMP‐9 (seen at 92 KDa) decreases after MCP‐1 treatment. In contrast, MMP2 activity (seen at 72 KDa) is unaltered. (B) Quantitation of data. Band intensity was quantitated electronically, and changes in band intensity are shown. MCP‐1 decreased MMP‐9 band intensity to 38–53% relative to untreated cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4255798&req=5

fig01: Effect of MCP‐1 on PF supernatant MMP‐9 activity as determined by gelatin zymography. (A) Representative zymogram. Primary rat PF were isolated and either left untreated or were treated overnight with MCP‐1 (0.1–50 ng/mL). Cells were washed, and the cell supernatant was collected after 30 min. As can be seen, the relative activity of MMP‐9 (seen at 92 KDa) decreases after MCP‐1 treatment. In contrast, MMP2 activity (seen at 72 KDa) is unaltered. (B) Quantitation of data. Band intensity was quantitated electronically, and changes in band intensity are shown. MCP‐1 decreased MMP‐9 band intensity to 38–53% relative to untreated cells.
Mentions: We investigated the effect of MCP‐1 on MMP‐9 activity via gelatin zymography. As seen in Figure 1, MCP‐1 decreased MMP‐9 activity in PF in a concentration‐sensitive fashion. Note that no change in MMP‐2 activity was observed, consistent with the concept that, in general, MMP‐9 is regulated (Hatfield et al. 2010), whereas MMP2 is constitutive (Gioia et al. 2009). We then examined whether the observed downregulation of MMP‐9 activity was due to downregulation of transcription. As seen in Figure 2A, MCP‐1 induced no change in MMP‐9 mRNA. Finally, we investigated whether MCP‐1 regulated the intracellular protein content of MMP‐9. We found that MMP‐9 protein could be detected in PF only after treatment with MCP‐1 (10 or 100 ng/mL). Taken together, these findings suggested that MCP‐1 altered PF MMP‐9 activity in a fashion independent of transcriptional changes and possibly due to alteration of protein release into the extracellular environment.

Bottom Line: We examined the effect of MCP-1 on release of matrix metalloproteinase-9 (MMP-9) by rat PF.We found that MCP-1 blocks PF release of MMP-9 in a posttranslational fashion.Our data demonstrated that, in the presence of MCP-1, MMP-9-containing vesicles were shunted to a lysosome-like compartment.

View Article: PubMed Central - PubMed

Affiliation: Division of Gastroenterology and Hepatology, Department of Internal Medicine, University of Arkansas for Medical Sciences, Little Rock, Arkansas.

No MeSH data available.


Related in: MedlinePlus