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Objective The purpose of the study was to investigate the changes and clinical significance of matrix metalloproteinase (MMP)-1,MMP-3,tissue inhibitor of metalloproteinase (TIMP),interleukin (IL)-1,tumor necrosis factor (TNF)-α and transforming growth factor(TGF)-β in juvenile idiopathic arthritis (JIA).Methods Thirty-two JIA subjects and 28 controls (traumatic arthritis patients) were included into this study.The MMP-1,MMP-3,TIMP,IL-1,TNF-α and TGF-β level in the serum and synovia were assessed by ELISA.The WBC count,the level of CRP,ESR,RF were also detected.Independent t-test and Pearson's analysis were adopted for data analysis.Results ① The level of MMP-1,MMP-3,IL-1 and TNF-α in the serum was (158±67) ng/ml,(212±89) ng/ml,(39±19) pg/ml,(26±10) pg/ml respectively,which was significantly higher than that of the control group (all P＜0.05) ; the ratio of MMP-3/TIMP-1 (0.86±0.32) was higher in the study group than that of the control group (P＜0.05),while the value of TIMP-1,TGF-β was (248±88),(17±9) ng/ml respectively,which was lower than that of the control group (P＜0.05).The value of seral MMP-3,MMP-3/TIMP-1 was positively correlated with that of WBC,CRP,ESR in the JIA group (all P＜0.05).② The value of MMP-1,MMP-3,IL-1 and TNF-α in the synovia was (216±78) ng/nl,(766±291)ng/ml,(56±21) pg/ml,(36±14) pg/ml respectively,which was higher than that of the control group (all P＜0.05); the ratio of MMP-3/TlMP-1 (2.68±0.89) was higher than that of the control group (P＜0.05),while the value of TIMP-1,TGF-β was (286±88) ng/ml,(12±4) ng/ml respectively,which was lower than that of the control group (all P＜0.05).③ The value of MMP-1,MMP-3,TIMP-1,IL-1 and TNF-α in the synovia was higher than that in the serum (all P＜0.05),while the value of TGF-β was lower than that in the serum (P＜0.05).Conclusion The value of MMP-1,MMP-3,IL-1 and TNF-α increases both in the serum and synovia,while the value of TIMP-1 decreases.The value of TGF-β decreases,which may have protective effect on JIA.The ratio of MMP-3/TIMP-1 in the serum is positively correlated to inflammation parameters,which may be used to judge the activity of illness in JIA.
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Objective To study whether p38 mitogen-activated protein kinases (p38MAPK) signal pathway were activated in the process of tumor necrosis factor-like weak inducer of apoptosis (TWEAK) which then induces rheumatoid arthritis (RA) fibroblast-like synoviocyte (FLS) to synthesize matrix metalloproteinase-1 (MMP-1) and look for the relative mechanisms of how TWEAK was involved in the destruction of articular bones and cartilage. Methods RA FLS were primarily cultured and stimulated with TWEAK. FLS were pretreated with SB203580 or not. ELISA was used to detect the concentration of MMP-1 in cell-cultured supernatant. Western blotting was used to detect the expression of p-p38MAPK and P65 in RA FLS. Results TWEAK (100 ng/ml) could induce RA FIS to synthesize MMP-1. SB203580 could partially inhibite the expression of MMP-1 producted by RA FLS which was induced by TWEAK. TWEAK could make p38MAPK phosphorylated and increase the expression of P65 protein in the cell nucleus. Conclusion TWEAK induces RA FLS to synthesize MMP-1. In this process, p38MAPK signal transduction pathway is activated and then induce the expression of NF-κB.
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Objective To investigate the role of interstitial collagenase in the pathogenesis of acute lung injury induced by hyperoxia outside of sealed cages and breath room air,and to study the mechanism of The severity of lung injury.Methods Seventy-two C57BL/6 mice were divided into normal control group,hyperoxia for 24 hours group,hyperoxia for 48 hours and hyperoxia for 72 hours group randomly,18 mice in each group.The hyperoxia group exposedin sealed cages with＞95%oxygen,and the control group were put in the inspiratory room.The expression of interstitial collagenase mRNA and protein in lung tissues was studied by reverse transcript-polymerase chain reaction(RT-PCR)and immunohistochemistry.Results Hyperoxia caused acute lung injury in mice.by The expression of interstitial collagenase mRNA in lung tissues was increased after 24 hours of hyperoxia compared with their control group[0.59±0.11 vs 0.07±0.01,q=3.t5 P＜0.01],the expression was higher at 72 hours of hyperoxia(0.68±0.12,q=3.78 P＜0.01).Immunohistochemistry study showed interstitial collagenase protein was mainly expressed in cytoplasm of airway epithelial cells,while Ⅱ type alveolar epithelial cells mainly and vascular smooth muscle cells in hyperoxia mice.The expression of interstitial collagenase protein in airway epithelium significantly increased at 24 hours of hyperoxia compared with their control group[(28.54±9.60) vs (13.48±4.32)q=2.62 P＜0.05],and the expression level was lower after 48 and 72 hours of hyperoxia(20.32±5.68) vs, (15.24±4.65).Conclusion Hyperoxia cause acute lung injury in mice;interstitial collagenase play an important role in the development of hyperoxia-induced lung injury in mice.
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Objective To study the effects of the IL-1 receptor antagonist (IL-1Ra) on matrix metalloproteinase 1 (MMP-1) expression in UV-irradiated fibroblasts. Methods Simulating the impact of environmental ultraviolet (UV) light on human skin, UVA-irradiated human fibroblasts were cultured in medium obtained from UVB-irradiated HaCaT cells. MMP-1 was detected by ELISA in the culture medium of fibroblasts. After treatment with IL-1Ra, the mRNA expression levels of C-Jun, C-Fos and GAPDH (internal control) of fibroblasts were measured by real-time fluorescent quantitative RT-PCR. Results Production of MMP-1 by UVA (10 J/cm2)-irradiated fibroblasts was increased in culture medium from UVB-irradiated HaCaT cells. The fibroblasts produced significantly higher levels of MMP-1 in culture medium from HaCaT cells treated without UVB than those with 15 mJ/cm2 UVB (t = 8.413,P= 0.014). However, IL-1Ra inhibited MMP-1 production of fibroblasts in a dose-dependent manner. Standard curves of real-time fluorescent quantitative RT-PCR showed a linear correlation between the copy number and the threshold cycle (Tc). Melting curves confirmed the specificity of PCR products. The original copy numbers of C-Jun and C-Fos as well as the ratios of the numbers to the GAPDH copy number showed that IL-1Ra inhibited the C-Jun mRNA expression of fibroblasts in a dose-dependent manner but had no significant effects on C-Fos mRNA expression. Conclusions The culture medium from UVB-irradiated HaCaT cells can promote MMP-1 production by UVA-irradiated fibroblasts. IL-1Ra reduces MMP-1 production via inhibition of C-Jun mRNA expression.
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BACKGROUND: Matrix metalloproteinase (MMP)-1 and 3 are the most important degradating enzymes of the chondroid matrix. Chondrocytes may undergo apoptosis under various stimuli including nitric oxide (NO). We studied the expression rate and zone of MMP-1, MMP-3, nitrotyrosine, a marker of NO release, and apoptosis in the articular cartilage of human osteoarthritis. METHODS: To investigate the role of nitrotyrosine and apoptosis in the degradation of the chondroid matrix in human osteoarthritis, immunohistochemistry was done for MMP-1, MMP-3, and nitrotyrosine; and the terminal deoxynucleotidyl transferase mediated nick end labeling (TUNEL) method was performed for apoptosis using a total of 93 articular cartilages from 12 femoral heads and 17 knees obtained from total joint arthroplasty and 7 normal articular cartilages. RESULTS: In the normal control group, the expression rates for MMP-1, MMP-3, nitrotyrosine, and apoptosis were very low; and their expression zones were confined to the superficial layer of the articular cartilage. Their expression rates were low in the early stage of osteoarthritis and were moderate to high in the late stage (P<0.05). Their expression zones were confined to the superficial layer of the articular cartilage in the early stage of osteoarthritis and were expressed throughout the whole layer in the late stage and those of MMP-3 and nitrotyrosine were statistically significant (P<0.05). Their expression rates and zones were significantly correlated with the grade of osteoarthritis (P<0.05). Conclusion : The expression rate and zone of apoptosis and nitrotyrosine correlated well with those of MMP-1 and MMP-3. Therefore, NO and apoptosis may be related to the progression of human osteoarthritis.
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Humanos , Apoptosis , Artroplastia , Cartílago Articular , Condrocitos , ADN Nucleotidilexotransferasa , Cabeza , Inmunohistoquímica , Articulaciones , Rodilla , Metaloproteinasa 1 de la Matriz , Óxido Nítrico , OsteoartritisRESUMEN
In order to investigate the influence of interleukin 1?(IL 1?) and tumor necrosis factor ? (TNF ?) on gene expression of rat interstitial collagenase (MMP 13 ) in hepatic stellate cells, cells of a hepatic stellate cell line (HSC T6) were cultured in medium containing IL 1? or TNF ? for different incubation periods. Total RNA of HSC was extracted and MMP 13 gene expression levels were measured by reverse transcription polymerase chain reaction. The results showed that MMP 13 gene expression level of the IL 1? group was much higher than that of the control group at 8h, 24h and 48h after exposure to IL 1?. The maximal value was 3 fold of the control group and was at 24h . MMP 13 gene expression level of the TNF ? group was also significantly higher than that of control group at 8h, 24h, 48h, 72h, and the maximum nalue was 4 fold of the control group at 48h. These results indicated that IL 1? and TNF ? might increase the gene expression of MMP 13 in hepatic stellate cells.
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Objective To explore the fibrosis mechanism of patients with fibrosing cholestatic hepatitis(FCH) in the way of degradation of collagen.Methods The expressions of matrix metalloproteinase 1(MMP1) and tissue inhibitor of metalloproteinase 1(TIMP1),and contents of type I,III collagen proteins were detected by immunohistochemical staining in the liver tissues of 9 cases with FCH associated with HBV developed following renal transplantation and 5 cases without liver disease as controls.Results The expressions of MMP1 and TIMP1,and type I,III collagen proteins in the patients with FCH were significantly higher than those in the control group.There was a positive correlation between the expressions of type I,III collagen proteins and TIMP1/ MMP1 ratio.Conclusion Hepatic fibrosis in the patients with FCH associated with HBV developed following renal transplantation may be relative to the increase of TIMP1 expression which inhibit the degradation of collagen.
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Sclerosis is a disease process in which idiopathic hardening occurs in the skin and/or internal organs as a result of the accumulation of type I collagen, induced mainly by transforming growth factor-beta. Colchicine and D-penicillamine are widely used for its treatment. Their effects are known to be due to post-translational down-regulation of type I collagen synthesis, with colchicine also up-regulating interstitial collagenase. To determine whether or not they have any pre-translational effect on type I collagen and MMP-1, and also to observe their effects on the action of TGF-beta, cultured neonatal foreskin fibroblasts were treated with colchicine and D-penicillamine, singly and together. The amount of type I collagen and MMP-1 mRNA were quantitated by Northern blot hybridization. Colchicine suppresses the basal level of type I collagen mRNA but minimally stimulates the mRNA expression of MMP-1, whereas D-penicillamine does not have any significant effects on either. Colchicine was also able to significantly suppress the TGF-beta-induced up-regulation of type I collagen mRNA expression.