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Blockade of hypoxia-induced CXCR4 with AMD3100 inhibits production of OA-associated catabolic mediators IL-1β and MMP-13.

Li P, Deng J, Wei X, Jayasuriya CT, Zhou J, Chen Q, Zhang J, Wei L, Wei F - Mol Med Rep (2016)

Bottom Line: Binding of the chemokine stromal cell-derived factor-1 (SDF-1) to its receptor C-X-C chemokine receptor type 4 (CXCR4) results in receptor activation and the subsequent release of matrix metalloproteinases (MMPs) that contribute to osteoarthritis (OA) cartilage degradation.By contrast, such changes did not occur to an appreciable degree in cells that were pretreated with AMD3100.The results of the present study demonstrate that even under hypoxic conditions, where CXCR4 expression is significantly elevated in chondrocytes, AMD3100 effectively blocks this receptor and protects chondrocytes from OA‑induced catabolism, suggesting that the successful inhibition of CXCR4 may be an effective approach for OA treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedics, The Second Hospital of Shanxi Medical University, Shanxi Key Laboratory of Bone and Soft Tissue Injury Repair, Taiyuan, Shanxi 030001, P.R. China.

ABSTRACT
Binding of the chemokine stromal cell-derived factor-1 (SDF-1) to its receptor C-X-C chemokine receptor type 4 (CXCR4) results in receptor activation and the subsequent release of matrix metalloproteinases (MMPs) that contribute to osteoarthritis (OA) cartilage degradation. As hypoxia is a defining feature of the chondrocyte microenvironment, the present study investigated the possible mechanism through which SDF‑1 induces cartilage degradation under hypoxic conditions. To do this, OA chondrocyte cultures and patient tissue explants pretreated with the CXCR4 inhibitor, AMD3100 were incubated with SDF‑1. It was identified that hypoxic conditions significantly elevated the expression of CXCR4 in osteoarthritic chondrocytes relative to normoxic conditions. Furthermore, SDF‑1 elevated MMP‑13 mRNA levels and proteinase activity. It also elevated the mRNA and protein levels of runt‑related transcription factor 2, and induced the release of glycosaminoglycans and the inflammatory cytokine, interleukin‑1β. By contrast, such changes did not occur to an appreciable degree in cells that were pretreated with AMD3100. The results of the present study demonstrate that even under hypoxic conditions, where CXCR4 expression is significantly elevated in chondrocytes, AMD3100 effectively blocks this receptor and protects chondrocytes from OA‑induced catabolism, suggesting that the successful inhibition of CXCR4 may be an effective approach for OA treatment.

No MeSH data available.


Related in: MedlinePlus

SDF-1/CXCR4 signaling pathway regulates MMP-13 expression and activity. (A) SDF-1 induced MMP-13 activity in a dose-dependent manner. Cells were treated with SDF-1 in serial dosages of 10, 20, 40 and 80 ng/ml for two days and the conditioned media were collected for ELISA analysis. The increasing SDF-1 effect reached its highest point at the dosage of 40 ng/ml. *P<0.05 vs. control; #P<0.05 vs. cells treated with 40 ng/ml SDF-1; aP<0.05 vs. cells treated with 10 ng/ml SDF-1; bP<0.05 vs. cells treated with 20 ng/ml SDF-1; cP<0.05 vs. cells treated with 80 ng/ml SDF-1. (B) Reverse transcription-quantitative polymerase chain reaction analysis of MMP-13 expression in osteoarthritis chondrocytes treated with SDF-1. SDF-1 significantly increased MMP-13 mRNA expression level in cells (P=0.009), which was significantly reduced by blocking the SDF-1 pathway with AMD 3100 (P=0.001). (C) ELISA analysis of enzyme activity of MMP-13 within conditioned media. SDF-1 significantly increased the activity of MMP-13 (P=0.013), while a significant suppression of MMP-13 activity was observed by blocking the SDF-1 pathway with AMD 3100 (P=0.001). SDF-1, stromal cell-derived factor-1; CXCR4, C-X-C chemokine receptor type 4; MMP-13, matrix metalloproteinase 13.
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f3-mmr-14-02-1475: SDF-1/CXCR4 signaling pathway regulates MMP-13 expression and activity. (A) SDF-1 induced MMP-13 activity in a dose-dependent manner. Cells were treated with SDF-1 in serial dosages of 10, 20, 40 and 80 ng/ml for two days and the conditioned media were collected for ELISA analysis. The increasing SDF-1 effect reached its highest point at the dosage of 40 ng/ml. *P<0.05 vs. control; #P<0.05 vs. cells treated with 40 ng/ml SDF-1; aP<0.05 vs. cells treated with 10 ng/ml SDF-1; bP<0.05 vs. cells treated with 20 ng/ml SDF-1; cP<0.05 vs. cells treated with 80 ng/ml SDF-1. (B) Reverse transcription-quantitative polymerase chain reaction analysis of MMP-13 expression in osteoarthritis chondrocytes treated with SDF-1. SDF-1 significantly increased MMP-13 mRNA expression level in cells (P=0.009), which was significantly reduced by blocking the SDF-1 pathway with AMD 3100 (P=0.001). (C) ELISA analysis of enzyme activity of MMP-13 within conditioned media. SDF-1 significantly increased the activity of MMP-13 (P=0.013), while a significant suppression of MMP-13 activity was observed by blocking the SDF-1 pathway with AMD 3100 (P=0.001). SDF-1, stromal cell-derived factor-1; CXCR4, C-X-C chemokine receptor type 4; MMP-13, matrix metalloproteinase 13.

Mentions: The conditioned media were collected for ELISA analysis 48 h following treatment with SDF-1. As presented in Fig. 3A, SDF-1 (incubated at concentrations of 10-80 ng/ml) increased MMP-13 activity in a dose-dependent manner. Notably, the effect of SDF-1 was greatest at a dosage of 40 ng/ml. RT-qPCR and ELISA results indicated that SDF-1 increased the expression level of MMP-13 (P=0.009; Fig. 3B), while suppression of MMP-13 enzyme activity was observed by blocking the SDF-1 pathway with AMD3100 (P=0.001; Fig. 3C).


Blockade of hypoxia-induced CXCR4 with AMD3100 inhibits production of OA-associated catabolic mediators IL-1β and MMP-13.

Li P, Deng J, Wei X, Jayasuriya CT, Zhou J, Chen Q, Zhang J, Wei L, Wei F - Mol Med Rep (2016)

SDF-1/CXCR4 signaling pathway regulates MMP-13 expression and activity. (A) SDF-1 induced MMP-13 activity in a dose-dependent manner. Cells were treated with SDF-1 in serial dosages of 10, 20, 40 and 80 ng/ml for two days and the conditioned media were collected for ELISA analysis. The increasing SDF-1 effect reached its highest point at the dosage of 40 ng/ml. *P<0.05 vs. control; #P<0.05 vs. cells treated with 40 ng/ml SDF-1; aP<0.05 vs. cells treated with 10 ng/ml SDF-1; bP<0.05 vs. cells treated with 20 ng/ml SDF-1; cP<0.05 vs. cells treated with 80 ng/ml SDF-1. (B) Reverse transcription-quantitative polymerase chain reaction analysis of MMP-13 expression in osteoarthritis chondrocytes treated with SDF-1. SDF-1 significantly increased MMP-13 mRNA expression level in cells (P=0.009), which was significantly reduced by blocking the SDF-1 pathway with AMD 3100 (P=0.001). (C) ELISA analysis of enzyme activity of MMP-13 within conditioned media. SDF-1 significantly increased the activity of MMP-13 (P=0.013), while a significant suppression of MMP-13 activity was observed by blocking the SDF-1 pathway with AMD 3100 (P=0.001). SDF-1, stromal cell-derived factor-1; CXCR4, C-X-C chemokine receptor type 4; MMP-13, matrix metalloproteinase 13.
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getmorefigures.php?uid=PMC4940083&req=5

f3-mmr-14-02-1475: SDF-1/CXCR4 signaling pathway regulates MMP-13 expression and activity. (A) SDF-1 induced MMP-13 activity in a dose-dependent manner. Cells were treated with SDF-1 in serial dosages of 10, 20, 40 and 80 ng/ml for two days and the conditioned media were collected for ELISA analysis. The increasing SDF-1 effect reached its highest point at the dosage of 40 ng/ml. *P<0.05 vs. control; #P<0.05 vs. cells treated with 40 ng/ml SDF-1; aP<0.05 vs. cells treated with 10 ng/ml SDF-1; bP<0.05 vs. cells treated with 20 ng/ml SDF-1; cP<0.05 vs. cells treated with 80 ng/ml SDF-1. (B) Reverse transcription-quantitative polymerase chain reaction analysis of MMP-13 expression in osteoarthritis chondrocytes treated with SDF-1. SDF-1 significantly increased MMP-13 mRNA expression level in cells (P=0.009), which was significantly reduced by blocking the SDF-1 pathway with AMD 3100 (P=0.001). (C) ELISA analysis of enzyme activity of MMP-13 within conditioned media. SDF-1 significantly increased the activity of MMP-13 (P=0.013), while a significant suppression of MMP-13 activity was observed by blocking the SDF-1 pathway with AMD 3100 (P=0.001). SDF-1, stromal cell-derived factor-1; CXCR4, C-X-C chemokine receptor type 4; MMP-13, matrix metalloproteinase 13.
Mentions: The conditioned media were collected for ELISA analysis 48 h following treatment with SDF-1. As presented in Fig. 3A, SDF-1 (incubated at concentrations of 10-80 ng/ml) increased MMP-13 activity in a dose-dependent manner. Notably, the effect of SDF-1 was greatest at a dosage of 40 ng/ml. RT-qPCR and ELISA results indicated that SDF-1 increased the expression level of MMP-13 (P=0.009; Fig. 3B), while suppression of MMP-13 enzyme activity was observed by blocking the SDF-1 pathway with AMD3100 (P=0.001; Fig. 3C).

Bottom Line: Binding of the chemokine stromal cell-derived factor-1 (SDF-1) to its receptor C-X-C chemokine receptor type 4 (CXCR4) results in receptor activation and the subsequent release of matrix metalloproteinases (MMPs) that contribute to osteoarthritis (OA) cartilage degradation.By contrast, such changes did not occur to an appreciable degree in cells that were pretreated with AMD3100.The results of the present study demonstrate that even under hypoxic conditions, where CXCR4 expression is significantly elevated in chondrocytes, AMD3100 effectively blocks this receptor and protects chondrocytes from OA‑induced catabolism, suggesting that the successful inhibition of CXCR4 may be an effective approach for OA treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedics, The Second Hospital of Shanxi Medical University, Shanxi Key Laboratory of Bone and Soft Tissue Injury Repair, Taiyuan, Shanxi 030001, P.R. China.

ABSTRACT
Binding of the chemokine stromal cell-derived factor-1 (SDF-1) to its receptor C-X-C chemokine receptor type 4 (CXCR4) results in receptor activation and the subsequent release of matrix metalloproteinases (MMPs) that contribute to osteoarthritis (OA) cartilage degradation. As hypoxia is a defining feature of the chondrocyte microenvironment, the present study investigated the possible mechanism through which SDF‑1 induces cartilage degradation under hypoxic conditions. To do this, OA chondrocyte cultures and patient tissue explants pretreated with the CXCR4 inhibitor, AMD3100 were incubated with SDF‑1. It was identified that hypoxic conditions significantly elevated the expression of CXCR4 in osteoarthritic chondrocytes relative to normoxic conditions. Furthermore, SDF‑1 elevated MMP‑13 mRNA levels and proteinase activity. It also elevated the mRNA and protein levels of runt‑related transcription factor 2, and induced the release of glycosaminoglycans and the inflammatory cytokine, interleukin‑1β. By contrast, such changes did not occur to an appreciable degree in cells that were pretreated with AMD3100. The results of the present study demonstrate that even under hypoxic conditions, where CXCR4 expression is significantly elevated in chondrocytes, AMD3100 effectively blocks this receptor and protects chondrocytes from OA‑induced catabolism, suggesting that the successful inhibition of CXCR4 may be an effective approach for OA treatment.

No MeSH data available.


Related in: MedlinePlus