OBJECTIVETo investigate the degradation of artificial basement membrane (matrigel) co-cultured with oral carcinoma-associated fibroblasts (CAFs) and its possible mechanism.

METHODSCAFs and normal fibroblasts (NFs) were incubated on matrigel for 24, 48, 72 h. Equivalent amounts of conditioned medium were collected and assayed for total protein, hydroxyproline and matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) activity by gelatin zymography.

RESULTSOral CAFs were superior to oral NFs in total protein and hydroxyproline density, CAFs present more pro-MMP-2 and activated MMP-2.

CONCLUSIONCAFs were superior to NFs in degradation of matrigel. CAFs might play a key role in the reconstitution of extracellular matrix and the progression of tumor.

Basement Membrane ; Coculture Techniques ; Enzyme Precursors ; Fibroblasts ; Gelatinases ; Humans ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinase 9 ; Membranes, Artificial ; Mouth Neoplasms

PURPOSE: To evaluate the expression pattern of gelatinases in the human retinal pigment epithelial cell line ARPE-19 in response to oxidative stress. METHODS: ARPE-19 cells were exposed to paraquat for 72 hours. The cells were assayed for the mRNA expression of gelatinase A and gelatinase B by reverse transcriptase polymerase chain reaction (RT-PCR). The conditioned media were assayed for the production of gelatinases protein by Western blotting. Also, the inhibitory effect of the antioxidants against oxidative stress to the cells was evaluated. RESULTS: The gelatinase levels in the RPE cells increased under oxidative stress compaired with the control (p<0.05). Also, the levels decreased when the cells were incubated with the antioxidant N-acetylcysteine (NAC). CONCLUSIONS: Oxidative stress upregulates gelatinase expression in ARPE-19 cells.
Acetylcysteine ; Antioxidants ; Blotting, Western ; Culture Media, Conditioned ; Epithelial Cells ; Gelatinases* ; Humans ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinase 9 ; Oxidative Stress* ; Paraquat ; Retinaldehyde ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Messenger

OBJECTIVETo determine whether expression and activity of atrial gelatinases are altered in patients with atrial fibrillation (AF).

METHODSThe right atrial tissue samples were taken from 75 patients with rheumatic heart disease who underwent heart valve replacement surgery. 34 patients were in sinus rhythm, 11 patients had paroxysmal AF and 30 patients had persistent AF. The mRNA and protein level of MMP-2, MMP-9, TIMP-1, TIMP-2 were measured by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western-blotting analysis respectively. The activity of MMP-2 and MMP-9 was measured by zymographic analysis.

RESULTS(1) The mRNA level of MMP-2 increased significantly in the persistent AF group followed by the paroxysmal AF group compared with the sinus rhythm group (P < 0.01, respectively). MMP-9 mRNA expression remained compatible within groups (P > 0.05). MMP-2 and MMP-9 protein expression was prominent in the persistent AF group compared with the sinus rhythm and paroxysmal AF groups (P < 0.01), the significant difference was also observed between the paroxysmal AF and sinus groups (P < 0.05). (2) TIMP-1 and TIMP-2 expression at mRNA and protein level were all down-regulated in the persistent AF group compared with the sinus rhythm group (P < 0.05), however, the trends of reduction did not reach statistical significance in the paroxysmal AF group (P > 0.05) except that of the mRNA level of TIMP-2 (P < 0.05). (3) The activity of MMP-2 and MMP-9 significantly increased in both paroxysmal AF and persistent AF groups compared with the sinus rhythm group (P < 0.05). The significant difference in MMP-9 was also observed between the persistent AF and paroxysmal AF groups (P < 0.01). (4) MMP-2 and MMP-9 expression at mRNA and protein level were positively correlated with left atrial dimension and AF duration (P < 0.05) and were negatively correlated with the mRNA and protein level of TIMP-2 and TIMP-1 respectively (P < 0.01).

CONCLUSIONSThe upregulation of MMP-2,9 gene expression and activity, along with the selective downregulation of TIMP-1,2 may have contributed to the atrial structural remodeling during AF through influencing collagen metabolism.

Adolescent ; Adult ; Atrial Fibrillation ; genetics ; metabolism ; Female ; Gelatinases ; genetics ; metabolism ; Gene Expression ; Heart Atria ; metabolism ; Humans ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Middle Aged ; RNA, Messenger ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; genetics ; metabolism ; Young Adult

PURPOSE: Bronchial asthma is an inflammatory respiratory disease characterized by the activation of inflammatory cells and its infiltration. It has been recently reported that MMP- 9 dose an importance role in the movement of inflammatory cells through basal membrane, that the function may be suppressed by TIMP-1. We studied to know the change of MMP-9 and TIMP-1 in sputum before and after corticosteroid (CS) therapy, and the relation with MMP-9/TIMP-1 ratio and improvement of FEV1. METHODS: Seventeen acute moderate to severe asthmatics were selected as was a control group of 17 healthy children. MMP-9 and TIMP-1 in sputum were measured on the 0 day, 7 days and 3 months later and observed as to the flow of time. FEV1 was measured before the CS therapy and 3 months later, and the change of FEV1 & FEV1 at 3 months were compared with the relation of MMP-9/TIMP-1 ratio. RESULTS: Sputum MMP-9 was lowered more at 7 days and 3 months compared with 0 day (P< 0.05). Sputum TIMP-1 was significantly high on 7 days (P< 0.05) and then had a tendency to decrease until 3 months (P< 0.05). MMP-9/TIMP-1 ratio decreased according to the flow of time (P< 0.05). MMP-9/TIMP-1 ratio at 3 months closely correlated with the change of FEV1 (r=0.65, P< 0.05). CONCLUSION: These data suggest that the overproduction of MMP-9 after asthma exacerbation correlates with airway inflammation and TIMP-1 production might contribute to airway fibrosis. MMP-9/TIMP-1 ratio at 3 months correlates with improvement of pulmonary function after CS therapy.
Asthma* ; Child ; Fibrosis ; Gelatinases ; Humans ; Inflammation ; Matrix Metalloproteinase 1* ; Matrix Metalloproteinase 9* ; Membranes ; Prednisolone ; Sputum* ; Tissue Inhibitor of Metalloproteinase-1

OBJECTIVETo study the modulation effect of Naomaitong on gelatinase system after cerebral ische-Focal cerebral I/R rat model was duplicated by method of the intralumimia/reperfusion (I/R) in rats.

METHODnal filament technique. Rats were randomly divided into the sham-operative group, the model group, the Naomaitong group and the Nimodipine group, the latter three groups were also divided into the 3 hrs after ischemia group, and 6 hrs, 12 hrs, 24 hrs, 3 d, 6 d after IR groups. Immunohistochemical method and zymogram analysis method, etc. were adopted to observe the change of microvessel structure, gelatinase and its inhibitor expression.

RESULTSMMP-2 (IR 24 h-6 d)and MMP-9 (I/R 12 h-3 d) expression levels could be lowered and TIMP-1 expression level (IR 24 h-6 d) improved by Naomaitong. Besides, comparison of MMP-2 and MMP-9 content in zymogram analysis in each group showed that changes of its quantity were in accordance with the laws of immune expression.

CONCLUSIONThe protective effect of Naomaitong on cerebromicrovessel basement membrane injury in rats is related to its modulation on gelatinase system.

Animals ; Basement Membrane ; drug effects ; metabolism ; Brain Ischemia ; enzymology ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Gelatinases ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Medicine, Chinese Traditional ; Microvessels ; drug effects ; metabolism ; Protective Agents ; pharmacology ; Rats ; Reperfusion Injury ; enzymology ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism

OBJECTIVE: To evaluate whether lipopolysaccharide (LPS) modulates the expression of cyclooxy- genase-2 (COX-2) and also whether COX-2 is involved in the LPS induced matrix metalloproteinase-2 (MMP-2) and MMP-9 activation in human trophoblastic (TL) cell line. METHODS: We used the TL (trophoblast-like) cells and evaluated the effect of LPS on expression of COX-2 mRNA and protein and on activities of MMP-2 and MMP-9. Also, we pretreated cell line with LPS and NS398, a COX-2 inhibitor, and compared MMPs activities with LPS only group. In the present study, COX-2 was analyzed by RT-PCR and western blot analysis and gelatin zymography was done for the evaluation of gelatinase activities of MMP-2 and MMP-9. RESULTS: The mRNA and protein expressions of COX-2 were increased by LPS in time- and dose-dependant fashions. COX-2 mRNA expression began to rise from 1 hour of LPS treatment and was increased steadily thereafter. COX-2 protein expression was detected from 1 hour of LPS treatment, but maximally increased by the 3 hours of treatment. LPS also increased MMP-2 and MMP-9 activities in time and dose dependant fashions. Especially, active form of MMP-9 was observed in the high concentration of LPS (>50 microgram/ml). When adding COX-2 inhibitor (NS398) to LPS pretreated cell line, the MMPs activities increased in two or three fold compared to LPS only group. CONCLUSION: Our results suggested that LPS induces expression COX-2 and up-regulates activities of MMP-2 and MMP-9 in trophoblastic cell, but COX-2 although involved in LPS mediated MMP-2 and MMP-9 activation, may act through a different pathway than the commonly known prostaglandin metabolites mediated one.
Blotting, Western ; Cell Line* ; Gelatin ; Gelatinases ; Humans* ; Matrix Metalloproteinase 2* ; Matrix Metalloproteinases ; RNA, Messenger ; Trophoblasts*

Osteoarthritis is recognised to be an interactive pathological process involving the cartilage, subchondral bone and synovium. The signals from the synovium play an important role in cartilage metabolism, but little is known regarding the influence of the signalling from bone. Additionally, the collagenases and stromelysin-1 are involved in cartilage catabolism through mitogen-activated protein kinase (MAPK) signalling, but the role of the gelatinases has not been elucidated. Here, we studied the influence of osteoclastic signals on chondrocytes by characterising the expression of interleukin-1β (IL-1β)-induced gelatinases through MAPK signalling. We found that osteoclast-conditioned media attenuated the gelatinase activity in chondrocytes. However, IL-1β induced increased levels of gelatinase activity in the conditioned media group relative to the mono-cultured chondrocyte group. More specifically, IL-1β restored high levels of gelatinase activity in c-Jun N-terminal kinase inhibitor-pretreated chondrocytes in the conditioned media group and led to lower levels of gelatinase activity in extracellular signal-regulated kinase or p38 inhibitor-pretreated chondrocytes. Gene expression generally correlated with protein expression. Taken together, these results show for the first time that signals from osteoclasts can influence gelatinase activity in chondrocytes. Furthermore, these data show that IL-1β restores gelatinase activity through MAPK inhibitors; this information can help to increase the understanding of the gelatinase modulation in articular cartilage.
3T3 Cells ; Animals ; Cartilage, Articular ; cytology ; Cell Survival ; physiology ; Cells, Cultured ; Chondrocytes ; drug effects ; enzymology ; Coculture Techniques ; Culture Media, Conditioned ; Gelatinases ; drug effects ; Interleukin-1beta ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; MAP Kinase Signaling System ; physiology ; Matrix Metalloproteinase 2 ; drug effects ; Matrix Metalloproteinase 9 ; drug effects ; Mice ; Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; drug effects ; Monocytes ; cytology ; NF-kappa B ; antagonists & inhibitors ; Osteoclasts ; physiology ; Protease Inhibitors ; analysis ; Tissue Inhibitor of Metalloproteinase-1 ; drug effects ; Tissue Inhibitor of Metalloproteinase-2 ; drug effects ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors

OBJECTIVE: To investigate the effect of form-deprivation on level of gelatinase in the posterior sclera in chicks. METHODS: Fifty 1-day-old chicks were monocularly deprived to establish the animal model of form-deprivation myopia (FDM). According to the duration of form-deprivation the experimental chicks were divided randomly and equivalently into 5 groups, which were deprived for 3, 7, 14, 21 and 30 days respectively. Meanwhile the other eyes of the deprived chicks were used as self-control groups and chicks of the same days were chosen randomly as the normal control groups for each FDM group. At each form-deprivation point the changes of degree of diopters and axial length of chicks in each group were recorded. The levels of gelatinase in posterior sclera of the experimental eyes were measured by gelatin enzymography. RESULTS: Compared with the normal and self-control groups, the levels of MMP-2 activity in FDM groups were much higher (P <0.01). With the increase of the time of monocular deprivation these changes became more significant and reached the top after 14 days' deprivation with an inter-group statistical difference (P <0.01). The dynamic changes of MMP-2 activity were the same as those of axial length and degree of diopters in each experimental groups. There was positive correlation between the MMP-2 activity and axial length (r = 0.989, P < 0.01). But there was a negative correlation between the MMP-2 activity and refractive degree. CONCLUSION Increase of MMP-2 activity in the posterior sclera of chicks would be a direct key factor to trigger sclera ECM remodeling process in chick FDM.
Animals ; Chickens ; Gelatinases ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Myopia ; enzymology ; etiology ; Sclera ; enzymology

OBJECTIVETo investigate the changes in expression of gelatinase (MMP-2, MMP-9) in young rat condylar cartilage during functional mandibular advancement.

METHODSSixty male 5-week old SD rats were divided into experimental and control groups. The mimic functional appliances were used in experimental group rats. The animals were sacrificed after 1, 2, 4 weeks. The immunoreactivity of gelatinase was detected by immunohistochemistry.

RESULTSIn normal mandibular condylar cartilage, the immunoreactivity of MMP-2 was rather strong, the immunoreactivity of MMP-9 was very weak. Following functional mandibular advancement, the expression of MMP-9 was significantly increased (P < 0.01), but immunoreactivity of MMP-2 had no significant changes.

CONCLUSIONGelatinase plays an important role in the adaptive remondling of young rat condylar cartilage during functional mandibular advancement.

Animals ; Cartilage ; Gelatinases ; Male ; Mandibular Advancement ; Mandibular Condyle ; Matrix Metalloproteinase 2 ; Rats ; Rats, Sprague-Dawley

BACKGROUND: MMPs and TIMPs are important factors for abnormal remodeling the pulmonary parenchyme in idiopathic interstitial pneumonia(IIP) This study evaluated the expression of MMPs and TIMPs in the tissue of IPF, NSIP and normal control subjects. METHOD: The MMP-2 and -9 activity in the lung tissue was studied by gelatin zymography, and the expression of MMP-1, -2 ,-9, TIMP-1 and -2 in the lung tissue was measured by immunohistochemistry. Thirty five patients, who were diagnosed with IIP (UIP ; 22, NSIP ; 13), were enrolled in the immunohistochemical study. Thirteen patients with IIP (UIP ; 9, NSIP ; 4) and five patients with lung cancer were enrolled in the zymographic assay. RESULTS: (1) The immunohistochemistry for MMP-1,-2,-9, TIMP-1 and-2 ; MMP-1,-9 and TIMP-2 were stained stronger in the UIP subjects than NSIP and the normal control. TIMP-2 was strongly stained in the UIP tissue. particularly the fibroblasts in the fibroblastic foci. (2) Zymography for MMP-2 and MMP-9 revealed MMP-2 to have prominent expression in the UIP tissue than in the NSIP tissue. CONCLUSIONS: These results suggest that the overexpression of the TIMPs and gelatinases in UIP might be? important factors in the irreversible fibrosis of the lung parenchyme.
Fibroblasts ; Fibrosis ; Gelatin ; Gelatinases ; Humans ; Immunohistochemistry ; Lung ; Lung Neoplasms ; Matrix Metalloproteinases* ; Tissue Inhibitor of Metalloproteinase-1 ; Tissue Inhibitor of Metalloproteinase-2