Inhibitory effect of midazolam on MMP-9, MMP-1 and MMP-13 expression in PMA-stimulated human chondrocytes via recovery of NF-κB signaling

Jen Jui Wang, Steven Kuan Hua Huan, Kuo Hsien Hsieh, Hsiu Chu Chou, George Hsiao, Thanasekaran Jayakumar, Joen Rong Sheu

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Introduction: Midazolam, a benzodiazepine, has a hypnotic effect and is widely used as an intravenous sedative. Past studies have clearly established that midazolam has beneficial effects in attenuating ischemia-reperfusion injury more than other currently used sedative drugs. However, the role of midazolam on chondroprotection via inhibition of matrix metalloproteinases (MMPs) is warrant investigation. The aim of this study was to examine the mechanisms of action of midazolam on MMP expression via nuclear factor κB (NF-κB) signaling in activated chondrosarcoma cells maintained in vitro. Material and methods: Chondrocytes, SW1353 cells, were stimulated with phorbol 12-myristate 13-acetate (PMA) in the absence or presence of various concentrations of midazolam (5-20 μM). Release of MMP-9 into the culture media was determined by gelatin zymography. The expressions of MMP-1, MMP-9 and MMP-13, phosphorylation of extracellular signal-regulated kinase (ERK), p38 mitogen-activated protein kinases and degradation of IκB-α were determined by western blotting assay. Results: Midazolam significantly down-regulated PMA-induced MMP-9 protein expression at concentrations of 5, 10 and 20 μM, the values were 1.95 ±0.09 (p <0.01), 1.71 ±0.12 (p <0.01) and 1.35 ±0.20 (p <0.001), respectively. At concentrations of 5, 10 and 20 μM, it was significantly inhibited the PMA-induced expressions of MMP-1 (2.27 ±0.10, 1.98 ±0.11 and 1.56 ±0.15; p <0.001) and MMP-13 (0.89 ±0.04, 0.81 ±0.07, and 0.74 ±0.09; p <0.001), respectively. Midazolam at concentrations of 10 and 20 μM for 15 min significantly reversed the rate of degradation (0.895 ±0.051; p <0.05 and 0.926 ±0.060; p <0.01, respectively) of IκB-α in PMA-chondrocyte cells. In addition, this sedative drug inhibited PMA-induced levels of phos-ERK (1.243 ±0.12, 1.108 ±0.16 and 0.903 ±0.19, respectively) and phos-p38 (1.146 ±0.10, 1.063 ±0.13 and 0.946 ±0.18, at concentrations of (5, 10 and 20 μM), respectively. Conclusions: These results are important for understanding the mechanism of midazolam in inhibiting PMA-induced MMP expression through the signaling pathways of either NF-κB or ERK/p38 MAPKs down-regulation.

Original languageEnglish
Pages (from-to)332-339
Number of pages8
JournalArchives of Medical Science
Volume9
Issue number2
DOIs
Publication statusPublished - Apr 2013

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Matrix Metalloproteinase 13
Matrix Metalloproteinase 1
Midazolam
Matrix Metalloproteinase 9
Chondrocytes
Acetates
Hypnotics and Sedatives
Extracellular Signal-Regulated MAP Kinases
Matrix Metalloproteinases
p38 Mitogen-Activated Protein Kinases
Chondrosarcoma
phorbol-12-myristate
Gelatin
Reperfusion Injury
Benzodiazepines
Pharmaceutical Preparations
Proteolysis
Culture Media
Down-Regulation
Western Blotting

Keywords

  • Human chondrosarcoma cells
  • IκB-α.
  • Matrix metalloproteinases
  • Midazolam
  • Mitogen-activated protein kinases
  • Phorbol 12-myristate 13-acetate

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Inhibitory effect of midazolam on MMP-9, MMP-1 and MMP-13 expression in PMA-stimulated human chondrocytes via recovery of NF-κB signaling. / Wang, Jen Jui; Huan, Steven Kuan Hua; Hsieh, Kuo Hsien; Chou, Hsiu Chu; Hsiao, George; Jayakumar, Thanasekaran; Sheu, Joen Rong.

In: Archives of Medical Science, Vol. 9, No. 2, 04.2013, p. 332-339.

Research output: Contribution to journalArticle

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abstract = "Introduction: Midazolam, a benzodiazepine, has a hypnotic effect and is widely used as an intravenous sedative. Past studies have clearly established that midazolam has beneficial effects in attenuating ischemia-reperfusion injury more than other currently used sedative drugs. However, the role of midazolam on chondroprotection via inhibition of matrix metalloproteinases (MMPs) is warrant investigation. The aim of this study was to examine the mechanisms of action of midazolam on MMP expression via nuclear factor κB (NF-κB) signaling in activated chondrosarcoma cells maintained in vitro. Material and methods: Chondrocytes, SW1353 cells, were stimulated with phorbol 12-myristate 13-acetate (PMA) in the absence or presence of various concentrations of midazolam (5-20 μM). Release of MMP-9 into the culture media was determined by gelatin zymography. The expressions of MMP-1, MMP-9 and MMP-13, phosphorylation of extracellular signal-regulated kinase (ERK), p38 mitogen-activated protein kinases and degradation of IκB-α were determined by western blotting assay. Results: Midazolam significantly down-regulated PMA-induced MMP-9 protein expression at concentrations of 5, 10 and 20 μM, the values were 1.95 ±0.09 (p <0.01), 1.71 ±0.12 (p <0.01) and 1.35 ±0.20 (p <0.001), respectively. At concentrations of 5, 10 and 20 μM, it was significantly inhibited the PMA-induced expressions of MMP-1 (2.27 ±0.10, 1.98 ±0.11 and 1.56 ±0.15; p <0.001) and MMP-13 (0.89 ±0.04, 0.81 ±0.07, and 0.74 ±0.09; p <0.001), respectively. Midazolam at concentrations of 10 and 20 μM for 15 min significantly reversed the rate of degradation (0.895 ±0.051; p <0.05 and 0.926 ±0.060; p <0.01, respectively) of IκB-α in PMA-chondrocyte cells. In addition, this sedative drug inhibited PMA-induced levels of phos-ERK (1.243 ±0.12, 1.108 ±0.16 and 0.903 ±0.19, respectively) and phos-p38 (1.146 ±0.10, 1.063 ±0.13 and 0.946 ±0.18, at concentrations of (5, 10 and 20 μM), respectively. Conclusions: These results are important for understanding the mechanism of midazolam in inhibiting PMA-induced MMP expression through the signaling pathways of either NF-κB or ERK/p38 MAPKs down-regulation.",
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author = "Wang, {Jen Jui} and Huan, {Steven Kuan Hua} and Hsieh, {Kuo Hsien} and Chou, {Hsiu Chu} and George Hsiao and Thanasekaran Jayakumar and Sheu, {Joen Rong}",
year = "2013",
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T1 - Inhibitory effect of midazolam on MMP-9, MMP-1 and MMP-13 expression in PMA-stimulated human chondrocytes via recovery of NF-κB signaling

AU - Wang, Jen Jui

AU - Huan, Steven Kuan Hua

AU - Hsieh, Kuo Hsien

AU - Chou, Hsiu Chu

AU - Hsiao, George

AU - Jayakumar, Thanasekaran

AU - Sheu, Joen Rong

PY - 2013/4

Y1 - 2013/4

N2 - Introduction: Midazolam, a benzodiazepine, has a hypnotic effect and is widely used as an intravenous sedative. Past studies have clearly established that midazolam has beneficial effects in attenuating ischemia-reperfusion injury more than other currently used sedative drugs. However, the role of midazolam on chondroprotection via inhibition of matrix metalloproteinases (MMPs) is warrant investigation. The aim of this study was to examine the mechanisms of action of midazolam on MMP expression via nuclear factor κB (NF-κB) signaling in activated chondrosarcoma cells maintained in vitro. Material and methods: Chondrocytes, SW1353 cells, were stimulated with phorbol 12-myristate 13-acetate (PMA) in the absence or presence of various concentrations of midazolam (5-20 μM). Release of MMP-9 into the culture media was determined by gelatin zymography. The expressions of MMP-1, MMP-9 and MMP-13, phosphorylation of extracellular signal-regulated kinase (ERK), p38 mitogen-activated protein kinases and degradation of IκB-α were determined by western blotting assay. Results: Midazolam significantly down-regulated PMA-induced MMP-9 protein expression at concentrations of 5, 10 and 20 μM, the values were 1.95 ±0.09 (p <0.01), 1.71 ±0.12 (p <0.01) and 1.35 ±0.20 (p <0.001), respectively. At concentrations of 5, 10 and 20 μM, it was significantly inhibited the PMA-induced expressions of MMP-1 (2.27 ±0.10, 1.98 ±0.11 and 1.56 ±0.15; p <0.001) and MMP-13 (0.89 ±0.04, 0.81 ±0.07, and 0.74 ±0.09; p <0.001), respectively. Midazolam at concentrations of 10 and 20 μM for 15 min significantly reversed the rate of degradation (0.895 ±0.051; p <0.05 and 0.926 ±0.060; p <0.01, respectively) of IκB-α in PMA-chondrocyte cells. In addition, this sedative drug inhibited PMA-induced levels of phos-ERK (1.243 ±0.12, 1.108 ±0.16 and 0.903 ±0.19, respectively) and phos-p38 (1.146 ±0.10, 1.063 ±0.13 and 0.946 ±0.18, at concentrations of (5, 10 and 20 μM), respectively. Conclusions: These results are important for understanding the mechanism of midazolam in inhibiting PMA-induced MMP expression through the signaling pathways of either NF-κB or ERK/p38 MAPKs down-regulation.

AB - Introduction: Midazolam, a benzodiazepine, has a hypnotic effect and is widely used as an intravenous sedative. Past studies have clearly established that midazolam has beneficial effects in attenuating ischemia-reperfusion injury more than other currently used sedative drugs. However, the role of midazolam on chondroprotection via inhibition of matrix metalloproteinases (MMPs) is warrant investigation. The aim of this study was to examine the mechanisms of action of midazolam on MMP expression via nuclear factor κB (NF-κB) signaling in activated chondrosarcoma cells maintained in vitro. Material and methods: Chondrocytes, SW1353 cells, were stimulated with phorbol 12-myristate 13-acetate (PMA) in the absence or presence of various concentrations of midazolam (5-20 μM). Release of MMP-9 into the culture media was determined by gelatin zymography. The expressions of MMP-1, MMP-9 and MMP-13, phosphorylation of extracellular signal-regulated kinase (ERK), p38 mitogen-activated protein kinases and degradation of IκB-α were determined by western blotting assay. Results: Midazolam significantly down-regulated PMA-induced MMP-9 protein expression at concentrations of 5, 10 and 20 μM, the values were 1.95 ±0.09 (p <0.01), 1.71 ±0.12 (p <0.01) and 1.35 ±0.20 (p <0.001), respectively. At concentrations of 5, 10 and 20 μM, it was significantly inhibited the PMA-induced expressions of MMP-1 (2.27 ±0.10, 1.98 ±0.11 and 1.56 ±0.15; p <0.001) and MMP-13 (0.89 ±0.04, 0.81 ±0.07, and 0.74 ±0.09; p <0.001), respectively. Midazolam at concentrations of 10 and 20 μM for 15 min significantly reversed the rate of degradation (0.895 ±0.051; p <0.05 and 0.926 ±0.060; p <0.01, respectively) of IκB-α in PMA-chondrocyte cells. In addition, this sedative drug inhibited PMA-induced levels of phos-ERK (1.243 ±0.12, 1.108 ±0.16 and 0.903 ±0.19, respectively) and phos-p38 (1.146 ±0.10, 1.063 ±0.13 and 0.946 ±0.18, at concentrations of (5, 10 and 20 μM), respectively. Conclusions: These results are important for understanding the mechanism of midazolam in inhibiting PMA-induced MMP expression through the signaling pathways of either NF-κB or ERK/p38 MAPKs down-regulation.

KW - Human chondrosarcoma cells

KW - IκB-α.

KW - Matrix metalloproteinases

KW - Midazolam

KW - Mitogen-activated protein kinases

KW - Phorbol 12-myristate 13-acetate

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