• Biomolecules & Therapeutics
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• 제26권6호
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• pp.560-567
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• 2018

In the present study, we tried to examine whether resveratrol regulates the expression of matrix metalloproteinases (MMPs) through affecting nuclear factor-kappa B ($NF-{\kappa}B$) in articular chondrocytes. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcription-polymerase chain reaction (RT-PCR) was used to measure interleukin-${\beta}$ ($IL-1{\beta}$)-induced gene expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), ADAMTS-5 and type II collagen. Effect of resveratrol on $IL-1{\beta}$-induced secretion of MMP-3 was investigated in rabbit articular chondrocytes using western blot analysis. To elucidate the action mechanism of resveratrol, effect of resveratrol on $IL-1{\beta}$-induced $NF-{\kappa}B$ signaling pathway was investigated in SW1353, a human chondrosarcoma cell line, by western blot analysis. The results were as follows: (1) resveratrol inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5, but increased the gene expression of type II collagen; (2) resveratrol reduced the secretion of MMP-3; (3) resveratrol inhibited $IL-1{\beta}$induced activation (phosphorylation) of inhibitory kappa B kinase (IKK), and thus phosphorylation and degradation of inhibitory kappa $B{\alpha}$ ($I{\kappa}B{\alpha}$); (4) resveratrol inhibited $IL-1{\beta}$-induced phosphorylation and nuclear translocation of $NF-{\kappa}B$ p65. This, in turn, led to the down-regulation of gene expression of MMPs in SW1353 cells. These results suggest that resveratrol can regulate the expression of MMPs through affecting $NF-{\kappa}B$ by directly acting on articular chondrocytes.

• 대한의생명과학회지
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• 제15권4호
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• pp.343-347
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• 2009

Cytochalasin D (CD) is known as a disruptor of actin cytoskeleton architecture in chondrocytes. We have studied the role of CD in retinoic acid (RA) caused dedifferentiation and inflammation responses in rabbit articular chondrocytes. We have examined the effect of CD on RA induced dedifferentiation of chondrocytes. CD inhibited RA induced dedifferentiation determined by Western blot analysis and Alcian blue staining in rabbit articular chondrocytes. Also, CD additionally reduced inflammation response molecules such as cyclooxygenase-2 (COX-2) and prostaglandin $E_2$ ($PGE_2$) in RA treated cells. Treatment of CD reduced phosphorylation of p38 by treatment of RA. Inhibiton of p38kinase with SB203580 reduced expression of COX-2 and production of $PGE_2$ by treatment of CD in RA treated cells. But, Inhibiton of p38kinase with SB203580 did not any relationship with effect of CD on RA caused dedifferentiation. In summary, our results indicate that CD regulates RA reduced expression of COX-2 and production of PGE2 via p38kinase pathway.

• Physical Therapy Rehabilitation Science
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• 제2권1호
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• pp.21-26
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• 2013

Objective: The aim of this study was to investigate the effect of therapeutic ultrasound (US) of cell viability and inflammatory cytokine in rat articular chondrocyte cultures stimulated with lipopolysaccharide (LPS). Design: One group pretest-posttest design. Methods: Cultured chondrocytes were treated with US and/or LPS and assessed for viability, Tumor necrosis factor $(TNF)-{\alpha}$ and Interleukin (IL)-1 production. Results: Oxidative stress was induced in rat chondrocytes with LPS. The cell viability was decreased in chondrocytes after treatment with LPS. The viability revealed that low-intensity pulsed ultrasound (LIPUS) exerted no significant cytotoxicity in the rat chondrocyte. LIPUS inhibited decreased cell viability in the presence of LPS ($30{\mu}g/ml$) in a intensity dependent pattern at LIPUS (p<0.05). $TNF-{\alpha}$ production in the presence of LPS was also inhibited in a dose dependent manner (p<0.05 from $30mW/cm^2$). IL-1 production in the presence of LPS was inhibited as well (p<0.05 from $7.5mW/cm^2$). Conclusions: Our results demonstrate that US was the anti-inflammatory effect of chondrocytes. LIPUS may exert its anti inflammatory effects through inhibition of $TNF-{\alpha}$ and IL-1 synthesis. These results suggest that US have potential for use as a pain relief and reduce the articular destruction.

• 동의생리병리학회지
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• 제20권2호
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• pp.414-419
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• 2006

Prostaglandins biosynthesis and nitric oxide production have been implicated in the process of inflammation and osteoarthritis. And nitric oxide (NO) activated the MMPs responsible for PG degradation in articular chondrocytes. Therefore, we have studied the effects on anti-inflammation and analgesic by ethyl acetate fraction from 70% ethanol extract of Perilla Frutescens (EPF). EPF inhibited LPS plus inflammation-cytokines-induced proteoglycan (PG) degradation, matrix-metalloproteinase (MMP-2,9) expression in rabbit articular chondrocytes. Also, EPF have inhibitory effects on LPS or LPS plus inflammation cytokines-induced nitric oxide (NO) and PGE2 production in mouse macrophage andrabbit articular chondrocytes. These results suggest that EPF decreases PGE2, iNOS, MMPs activity and PG degradation in mouse macrophage and/or rabbit articular chondrocytes. In vivo, EPF was shown to have inhibitory effects on acetic acid-induced pain. The herbal extract with this profile, may have utility in the treatment of osteoarthritis.

• 대한의생명과학회지
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• 제15권2호
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• pp.113-118
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• 2009

Actin cytoskeletal architecture is believed to be a crucially important modulator of chondrocyte phenotype. 2DG(2-Dexoy-D-glucose) induces reorganization of actin cytoskeletal architecture in chondrocytes. In this study, we have investigated the effects of 2DG on dedifferentiation and inflammation via reorganization of cytoskeletal architecture in rabbit articular chondrocytes, with a focus on p38 kinase pathway. Treatment of 2DG alone reduced type II collagen and COX-2 expression in chondrocytes. But, 2DG reduced type II collagen was recovered by CD, disruptor of actin cytoskeletal architecture, whereas did not affect on COX-2 expression and production of $PGE_2$ compared with 2DG alone treated cells. Treatment of 2DG with JAS, inducer of cytoskeletal architecture polymerization, accelerated reduction of type II collagen expression and synthesis of proteoglycan but did not affect on COX-2 expression and production of $PGE_2$. Also, 2DG stimulated activation of p38 kinase. This result showed that 2DG regulates type II collagen but not cyclooxygenase-2 expression through reorganization of cytoskeletal architecture via p38 kinase pathway in rabbit articular chondrocytes.

• 생명과학회지
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• 제29권8호
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• pp.888-894
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• 2019

류마티스관절염(rheumatoid arthritis)과 골관절염(osteoarthritis) 같은 관절질환은 연골세포(chondrocytes) 감소와 관절연골(articular cartilage)의 분해를 수반한다. 최근, 연골세포의 활성과 연골 항상성(cartilage homeostasis)에 염증성 ROS (reactive oxygen species) burst 및 나이와 관련된 산화적 스트레스(oxidative stress)의 증가와 관련된 연구가 활발히 진행되고 있다. 본 연구는 관절연골로부터 분리한 연골세포의 노화 과정과 나이대별 관절연골에서 항산화 인자들(antioxidants)의 변화를 조사함으로써, 연골세포와 관절연골의 노화 과정 동안 산화적 스트레스로부터 조직을 보호하는 항산화 인자들의 역할을 규명하는데 목적이 있다. 쥐의 관절연골로부터 분리한 연골세포의 연속 계대배양을 통한 노화 과정에서 산화적 스트레스가 증가함을 관찰하였다. 그리고, 노화 유도한 연골세포는 세포내 총 glutathione (GSSG/GSH) 양과 항산화 효소 superoxide dismutase (SOD)와 heme oxygenase-1 (HO-1)의 발현이 증가하였다. 다음으로, 나이대별 쥐로부터 분리한 관절연골에서 항산화 인자의 발현을 분석하였다. 항산화 인자 glutathione의 양은 40주령에서 발현이 가장 높게 관찰되었으며 72주령에 다소 감소하였고, SOD와 HO-1의 발현은 나이대별로 현저히 증가되는 경향을 보였다. 이를 종합해 볼 때, 세포내 항산화 인자들은 과도한 양의 ROS에 반응하여 연골세포의 노화와 나이와 관련된 관절연골의 퇴화 과정에서 중요한 역할을 하는 것으로 보인다.

• 대한의생명과학회지
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• 제14권3호
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• pp.179-186
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• 2008

Matrix metalloproteinases (MMPs), also designated matrixins, hydrolyze components of the extracellular matrix. These proteinases playa central role in many biological processes, such as embryogenesis, normal tissue remodeling, wound healing, and angiogenesis, and in diseases such as atheroma, arthritis, cancer, and tissue ulceration. In previous data, disruption of the actin cytoskeleton by cytochalasin D (CD) inhibited NO-induced apoptosis, dedifferentiation, cyclooxygenase (COX)-2 expression, and prostaglandin $E_2$ production in chondrocytes cultured on plastic or during cartilage explants culture. In this study, we investigated the effects of the actin cytoskeleton architecture on MMP-2 expression and dedifferentiation by CD in rabbit articular chondrocytes. Rabbit articular chondrocytes were prepared from cartilage slices of 2-weeks-old New Zealand white rabbits by enzymatic digestion. CD was used as a disruptor of actin cytoskeleton. In this experiments measuring CD dose response, primary chondrocytes were treated with various concentrations of CD for 24h. The actin disruption was determined by immunostaining. MMP-2 expression levels were determined by immunoblot analysis and Reverse transcriptase-Polymerase chain reaction (RT-PCR) and MMP-2 activity was determined by gelatin zymography. We found that cell morphological change and up-regulation of MMP-2 expression by CD as determined via immunostaining, gelatin zymography and immunoblotting. Moreover, CD induced MMP-2 transcription was detected by RT-PCR. Also, CD-induced type II collagen expression was inhibited by MMP-2 inhibitor I treatment. Our results indicate that CD up-regulated MMP-2 activation causes dedifferentiation of articular chondrocyte.

• 생명과학회지
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• 제20권10호
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• pp.1443-1450
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• 2010

Honokiol은 작은 분자량을 갖는 중국의 약초인 'Magnolia officinalis'에서 추출한 리간드로서 중국과 일본에서 전통 의약제로 사용되어 왔다. 이전 연구에서는 honokiol이 다양한 종류의 암에서anti-angiogenic, anti-invasive, anti-proliferative activities 등의 다양한 효과가 있는 것으로 알려져 있다. 우리는 이전 연구에서 Nitric oxide donor인 sodium nitroprusside (SNP)에 의해서 토끼 무릎관절연골세포에서 세포사멸이 일어나는 것을 보고하였다. 본 연구에서는 무릎 연골세포에서 NO에 의해 유도되는 세포사멸이 honokiol 처리농도에 의존적으로 억제되는 것을 세포형태, MTT assay, Western blot analysis 그리고 FACS를 통해 확인할 수 있었다. 또한, honokiol은 SNP에 의해 유도되는 p53의 발현 및 pro-caspase-3의 활성 저해, DNA fragmentation을 억제하였다. Honokiol에 의한 세포사멸 억제는 PI-3K의 특정 저해제인 LY294002를 SNP와 honokiol과 함께 처리하였을 때 세포의 사멸이 증가하는 것을 확인할 수 있었다. 이는 honokiol이 PI-3K/AKT pathway를 통하여 NO가 유도하는 세포사멸을 억제하는 것을 의미한다.

• Journal of Animal Science and Technology
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• 제56권8호
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• pp.33.1-33.8
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• 2014

Background: Sox 9 is a major marker of chondrocyte differentiation. When chondrocytes are cultured in vitro they progressively de-differentiate and this is associated with a decline in Sox 9 expression. The active form of vitamin D, 1, 25 $(OH)_2D_3$ has been shown to be protective of cartilage in both humans and animals. In this study equine articular chondrocytes were grown in culture and the effects of 1, 25 $(OH)_2D_3$ upon Sox 9 expression examined. The expression of the transient receptor potential vanilloid (TRPV) ion channels 5 and 6 in equine chondrocytes in vitro, we have previously shown, is inversely correlated with de-differentiation. The expression of these channels in response to 1, 25 $(OH)_2D_3$ administration was therefore also examined. Results: The active form of vitamin D (1, 25 $(OH)_2D_3$ when administered to cultured equine chondrocytes at two different concentrations significantly increased the expression of Sox 9 at both. In contrast 1, 25 $(OH)_2D_3$ had no significant effect upon the expression of either TRPV 5 or 6 at either the protein or the mRNA level. Conclusions: The increased expression of Sox 9, in equine articular chondrocytes in vitro, in response to the active form of vitamin D suggests that this compound could be utilized to inhibit the progressive de-differentiation that is normally observed in these cells. It is also supportive of previous studies indicating that $1{\alpha}$, 25-dihydroxyvitamin $D_3$ can have a protective effect upon cartilage in animals in vivo. The previously observed correlation between the degree of differentiation and the expression levels of TRPV 5/6 had suggested that these ion channels may have a direct involvement in, or be modulated by, the differentiation process in vitro. The data in the present study do not support this.

• 대한한의학회지
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• 제27권1호
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• pp.220-228
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• 2006

Objectives : Interleukin-1 (IL-1)${\beta}$ in articular chondrocytes regulates differentiation, apoptosis, and inflammatory responses. It is still controversial, So, we investigated IL- $1{\beta}$ induces chondrocytes dedifferentiation and death. Also, we studied the role of the mitogen-activated protein kinase (MAPK) subtypes on IL-$1{\beta}$-induced dedifferentiation and apoptosis. Methods : To evaluation of dedifferentiation by chemokines of chondrocytes, we assessed such as proteoglycan, collagen, MMP-3 and MMP-13 by RT-PCR analysis. Also, to assess of apoptosis effect by chemokines, we measured annexin V/propidium iodode (PI) and sub G1 cells in chondrocytes by flowcytometric analysis Results : IL-$1{\beta}$ treatment did not affect activation of ERK-1/2, but stimulation of p38 kinase. Inhibition of phospho ERK-1/2 with PD98059 enhanced IL-1b-induced dedifferentiation, and apoptosis up to 13.5%, whereas inhibition of phospho p38 kinase with SB203580 inhibited dedifferentiation, and apoptosis. Conclusions : Our results indicate that SB203580, p38 kinase inhibitor, inhibits IL-$1{\beta}$-induced dedifferentiation, and apoptosis by the inhibition of type II collagen expression and proteoglycan synthesis of rabbit articular chondrocytes.