BACKGROUND: Matrix metalloproteinases (MMPs) have been implicated in the remodelling of extracellular matrix (ECM), including basement membrane. ECM remodelling is associated with pathological processes, including hepatic fibrosis, tumor invasion and metastasis. Tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 were known to inhibit MMP-9 and MMP-2, respectively. In the present study, we examined the expression of TIMP-1 and TIMP-2 in surgical specimen pairs of hepatocellular carcinoma and nontumoral liver and the correlation between their expression and clinicopathological characteristics. METHODS: The localization of both transcripts and protein of TIMP-1 and TIMP-2 was studied by using in situ hybridization and immunohistochemistry. RESULTS: TIMP-1 and TIMP-2 mRNA transcripts were found in tumor cells, hepatocyte, sinusoidal cells, endothelial cells and stromal cells. Signal intensity of TIMP-1 was stronger than that of TIMP-2. The results of immunohistochemical stainings were concordant with those obtained by in situ hybridization. Expression of TIMP-1 and TIMP-2 was observed in tumorous tissue, in nontumorous tissue and in the portions of the tumors adjacent to the capsules. However, a clear difference in TIMP-1 and TIMP-2 mRNA expression was not observed among the three tissue types. The intensity of TIMP-2 expression was generally weaker than that of TIMP-1, and the intensity of TIMP-1 and TIMP-2 mRNA expression did not correlate with variable clinicopathological characteristics. CONCLUSION: TIMPs was expressed in tumor cells and many cell types of the nontumoral liver. Further investigations for TIMPs' unknown functional role are needed.
Adult ; Aged ; Carcinoma, Hepatocellular/pathology ; Carcinoma, Hepatocellular/metabolism* ; Female ; Human ; Immunohistochemistry ; Liver Neoplasms/pathology ; Liver Neoplasms/metabolism* ; Male ; Middle Age ; RNA, Messenger/analysis ; Tissue Inhibitor-of Metalloproteinase-2/physiology ; Tissue Inhibitor-of Metalloproteinase-2/genetics* ; Tissue Inhibitor-of Metalloproteinase-2/analysis ; Tissue-Inhibitor of Metalloproteinase-1/physiology ; Tissue-Inhibitor of Metalloproteinase-1/genetics* ; Tissue-Inhibitor of Metalloproteinase-1/analysis

OBJECTIVETo evaluate the effects of treadmill running exercise of different intensity on early repair of full-thickness defects on the patellofemoral articular surface and the changes in the expressions of matrix metalloproteinase-3 (MMP-3) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in SD rats.

METHODSTwenty-four male SD rats with full-thickness defects on the patellofemoral articular surface were randomly assigned into sedentary control (SED) group and low-, moderate- and high-intensity running groups (LIR, MIR, and HIR groups, respectively). The running groups were trained on treadmill for 6 consecutive weeks. Blood samples were collected to detect serum MMP-3 and TIMP-1 levels using ELISA before and after the experiment, and the femoral trochlea were collected to assess tissue repair by gross appearance scoring and O Driscoll histological scoring with Safranine O-Fast Green staining and Toluidine blue staining.

RESULTSIn rats in SED group, the defect was filled with hyaline articular cartilage-like tissues, as compared to fibrous tissues in LIR and MIR groups and subchondral bone damage in HIR group. The SED group scored the highest and HIR group the lowest among the 4 groups in gross appearance scoring and O Driscoll histological scoring. No significant differences were found in MMP-3 or TIMP-1 levels among the groups before training (P>0.05), but after 6 weeks of training, serum MMP-3 and TIMP-1 levels differed significantly among the 4 groups (P<0.05), and all the 3 running groups had a significantly higher MMP-3 level than the control group (P<0.05). After the 6-week training, TIMP-1/MMP-3 ratio was significantly higher in SED group than in the 3 running groups, and was the lowest in HIR group.

CONCLUSIONBoth low- and moderate-intensity exercise failed to promote resurfacing of full-thickness cartilage defects on the patellofemoral articular surface in rats, and high-intensity exercise even induces subchondral bone damage. The expression of MMP-3 and TIMP-1 is related to exercise, and the TIMP-1/MMP-3 ratio reflects the extent of tissue repair.

Animals ; Cartilage, Articular ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 1 ; metabolism ; Matrix Metalloproteinase 3 ; metabolism ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Wound Healing

BACKGROUNDExposure of adult mice to more than 95% O2 produces a lethal injury by 72 hours. Nuclear factor kappa B (NF-κB) is a transcriptional factor that plays a key role in the modulation of cytokine networks during hyperoxia-induced acute lung injury (ALI). Osteopontin (OPN) is a phosphorylated glycoprotein produced principally by macrophages. Studies have reported that exogenous OPN can maintain the integrity of the cerebral microvascular basement membrane and reduce brain damage through inhibiting NF-κB activities in the brain after subarachnoid hemorrhage. However, it is not clear whether OPN can reduce lung injury during ALI by inhibiting transcriptional signal pathways of NF-κB and consequent inhibition of inflammatory cytokines. Thus we examined the effects and mechanisms of recombinant OPN (r-OPN) on ALI.

METHODSNinety-six mice were randomly divided into phosphate buffered saline (PBS) and r-OPN groups. Mice were put in an oxygen chamber (>95% O2) and assessed for lung injury at 24, 48, and 72 hours. Expressions of NF-κB, matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9), and tissue inhibitors of MMP-2 and MMP-9 (TIMP-1, TIMP-2) mRNA in lungs were examined with RT-PCR. Expression and distribution of NF-κB protein in lungs were measured with immunohistochemistry.

RESULTSExposure to hyperoxia for 72 hours induced more severe lung injury in the PBS group compared with the r-OPN group. Expression of NF-κB mRNA in the PBS group exposed to hyperoxia for 48 and 72 hours was significantly higher than the r-OPN group (P < 0.05). With 72-hour exposure, expression of TIMP-1 mRNA in the r-OPN group was significantly higher than that of the PBS group (P < 0.05). Expression of TIMP-2 mRNA in the r-OPN group at 48 and 72 hours was significantly higher than those in the PBS group (P < 0.05). After 72-hour exposure, expression of NF-κB protein in airway epithelium in the PBS group was significantly higher than that in the r-OPN group (P < 0.05).

CONCLUSIONr-OPN can inhibit the release and activation of MMPs through inhibition of the expression of NF-κB and promotion of the expression of TIMPs, and alleviate hyperoxia-induced ALI.

Acute Lung Injury ; genetics ; metabolism ; Animals ; Hyperoxia ; metabolism ; physiopathology ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Mice ; NF-kappa B ; genetics ; metabolism ; Osteopontin ; genetics ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; genetics ; metabolism

OBJECTIVETo investigate the effects of serum from smoking individuals or non-smoking individuals with periodontitis on Porphyromonas gingivalis (Pg) internalizing KB cells, and the expression of matrix metalloproteinase(MMP)-1, MMP-9, tissue inhibitor of metalloproteinase-1 (TIMP-1) in the culture supernatant of KB cells.

METHODSThe venous blood of 20 periodontitis patients' (10 smoking and 10 non-smoking) was extracted under the informed consent and centrifuged for serum. The smoking-individual serum (Y group) and non-smoking-individual (N group) serum were added to the model of Pg internalizing KB cells for 12 hours, plated on brain-heart infusion (BHI) and incubated anaerobically at 37 °C for 5 days. The colony forming units (CFU) of cell-invasive bacteria were estimated by colony counting. MMP-1, MMP-9 and TIMP-1 protein levels in culture supernatant were determined by enzyme-linked immunosorbent assay(ELISA) in the two groups following co-culture of Pg with KB cells for 12 hours.

RESULTSThe CFU were (11.2 ± 1.1)×10(4), (12.6 ± 1.2)×10(4), (44.7 ± 1.3)×10(4) CFU/ml when adding 200, 400, 800 µl Y-group serum to the model of Pg co-culture with KB cells and when the serum was extracted from N group, the CFU were (33.6 ± 1.4)×10(4),(38.9 ± 1.1)×10(4), (11.2 ± 1.2)×10(4) CFU/ml respectively. When 200, 400, 800 µl Y group-serum was added to co-culture fluid of Pg internalizing KB cells, the concentrations of MMP-1 secreted from KB cells were (107.2 ± 21.5), (165.9 ± 20.2), (434.4 ± 48.0) µg/L respectively, the concentrations of MMP-9 were (3.99 ± 0.29), (4.21 ± 0.61), (5.62 ± 0.47) µg/L respectively, the concentrations of TIMP-1 were (401.3 ± 12.7), (418.3 ± 28.5), (637.3 ± 37.3) µg/L. When the serum (200, 400, 800 µl) extracted from N group, the concentration of MMP-1 and MMP-9 secreted by KB cell were (77.6 ± 10.8), (84.7 ± 10.2) and (98.2 ± 9.7) µg/L and (3.84 ± 0.52), (4.02 ± 0.68), (4.25 ± 0.37) µg/L, respectively. The concentration of TIMP-1 were (67.3 ± 26.9) , (89.4 ± 22.7) and (78.2 ± 16.5) µg/L secreted by KB cells in the course of Pg internalized KB cell. With the increasing of Y group-serum, the more MMP-1, MMP-9 and TIMP-1 were secreted by KB cells(P < 0.05). When 800 µl Y group-serum was added compared with N group-serum to the Pg co-culture with KB model, the more MMP-1, MMP-9 and TIMP-1 were secreted by KB cells(P < 0.05), when 400 µl Y group-serum was added compared with N group-serum to the Pg co-culture with KB model, the more MMP-1 and TIMP-1 were secreted by KB cells (P < 0.05).

CONCLUSIONSThe smoking-serum might enhance Pg internalizing KB cells and enhance the expression of MMP-1, MMP-9 and TIMP-1 secreted from KB cells. The local microenvironment of smoking individual may contribute to the recurrence and progression of chronic periodontitis.

Coculture Techniques ; Humans ; KB Cells ; Matrix Metalloproteinase 1 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Porphyromonas gingivalis ; enzymology ; RNA, Messenger ; Serum ; Smoking ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism

OBJECTIVETo investigate the effects of simvastatin(Sim) and losartan(Los) on cardiac fibrosis and myocardial expression of MMP-2 mRNA, MMP-9 mRNA and TIMP-1 mRNA, TIMP-2 mRNA in pressure overloaded rat hearts.

METHODSThe pressure overload model was induced by descending aortic constriction (DAC) in rats. SD rats were randomized into 6 groups (n = 20 each): normol control group, control sham group, DAC group, Los group (DAC + Los, 5 mg/kg), Sim group (DAC + Sim, 2 mg/kg), Los + Sim group (DAC + Los + Sim, Los 5 mg/kg, Sim 2 mg/kg). Water, Los or Sim drug was administrated by gavage daily beginning from day 5 after operation for 30 days. Collagen was measured on Masson stained myocardial sections, and the level of MMP-2 mRNA, MMP-9 mRNA and TIMP-1 mRNA, TIMP-2 mRNA in left ventricle were detected by RT-PCR.

RESULTSCollagen volume fraction (CVF) in DAC group was significantly higher than the normal control and sham groups (P < 0.01) which could be significantly reduced by Los and Sim (P < 0.05), especially in DAC + Los + Sim group (P < 0.01). The levels of myocardial MMP-2 mRNA, MMP-9 mRNA and TIMP-1 mRNA, TIMP-2 mRNA were also significantly higher in DAC group than in normal control and sham groups (P < 0.01). Treatment Sim and Los alone and especially in combination significantly decreased the TIMP-1 mRNA, TIMP-2 mRNA expressions (P < 0.01) while MMP-2 mRNA, MMP-9 mRNA levels remained unchanged (P > 0.05).

CONCLUSIONUpregulation of myocardial MMP-2 mRNA, MMP-9 mRNA and TIMP-1 mRNA, TIMP-2 mRNA expressions might contribute to myocardial fibrosis in this model, Sim and Los significantly inhibited myocardial fibrosis possibly by downregulating myocardial TIMP-1 mRNA, TIMP-2 mRNA expressions in this model.

Animals ; Gene Expression Regulation ; Heart Failure ; metabolism ; Losartan ; pharmacology ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Simvastatin ; pharmacology ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism

OBJECTIVETo examine the expression of MMPs and TIMPs in pleomorphic adenoma of salivary gland and to investigate the relationship between the expression and the biological behaviour of the tumor.

METHODSTwenty-three cases of pleomorphic adenoma were divided into active type and common type according to their biological behavior. Immunohistochemistry for MMP-2, MMP-9, TIMP-1, TIMP-2 and gelatin zymography analysis were performed in these 23 cases and in 6 malignant and 6 benign salivary gland tumors.

RESULTSThe immunoreactivity of MMP-2 protein and MMP-2/TIMP-1, 2 ratio were significantly higher in active pleomorphic adenoma than in common pleomorphic adenoma (P = 0.028, P = 0.009, P = 0.045). The expression of active MMP-2, proMMP-9 and active MMP-9 were significantly higher in active pleomorphic adenoma than in common pleomorphic adenoma (P = 0.034, P = 0.021, P = 0.001). There was no significant difference in expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 between the salivary malignant tumor and active pleomorphic adenoma, also between the salivary benign tumor and common pleomorphic adenoma.

CONCLUSIONSThe expression of MMPs and TIMPs in active pleomorphic adenoma is similar to that in salivary carcinomas, and the expression in common pleomorphic adenoma also resembled to that in salivary adenoma. The expression of MMP-2, 9 and TIMP-1, 2 is related to the biological behavior of pleomorphic adenoma of salivary gland.

Adenoma, Pleomorphic ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Salivary Gland Neoplasms ; metabolism ; pathology ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism

OBJECTIVETo study the expression and significance of matrix metalloproteinase (MMP) 2, MMP-9, tissue inhibitor of metalloproteinase (TIMP)1 and TIMP-2 in nasal cavity squamous cell carcinoma and their signification.

METHODSFifty cases of epithelial carcinoma tissue and 50 cases of normal nasal tissue were detected for MMP-2, MMP-9, TIMP-1, TIMP-2 by immunohistochemistry technique (S-P), and their relationship between the expression of MMP-2, MMP-9, TIMP-1, TIMP-2 and some clinical symptoms were analyzed. The SPSS 12.0 software was used to analyze the data.

RESULTSThe positive ratio of expression of MMP-2 in 50 cases of epithelial carcinoma was 52.0% (26/50), which was significantly higher (χ(2) = 6.00, P < 0. 05) than those [28.0% (14/50)] in the normal nasal tissue. The positive ratio of expression of MMP-9 in 50 cases of epithelial carcinoma was 58.0% (29/50), which was significantly higher (χ(2) = 12.8, P < 0.05) than those [10.0% (5/50)] in the normal nasal tissue. The positive ratio of expression of TIMP-1 in two groups was 74.0% (37/50), 56.0% (28/50) respectively. There was no difference between two groups (χ(2) = 0.51, P > 0.05). The positive ratio of expression of TIMP-2 in two groups was 26.0% (13/50), 20% (10/50) respectively. There was no difference between two groups (χ(2) = 3.35, P > 0.05).

CONCLUSIONSThere was a close relationship between pathogenesis and development of nasal epithelial carcinoma and the expression of MMP-2, MMP-9 in the epithelial carcinoma tissues. Both MMP and TIMP, especially the unbalance of MMP and TIMP, have prognostic value in nasal cavity squamous cell carcinoma.

Carcinoma, Squamous Cell ; metabolism ; pathology ; Humans ; Lymphatic Metastasis ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Nasal Cavity ; pathology ; Neoplasm Staging ; Nose Neoplasms ; metabolism ; pathology ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism

OBJECTIVETo observe the effects of hemoperfusion on oxidative stress status and the levels of matrix metallo proteinase (MMP-2, MMP-9), tissue inhibitor of metalloproteinase (TIMP-1) in lungs, livers and kidneys in paraquat poisoning rabbits, and to explore the mechanism of therapeutic effects induced by HP on acute paraquat poisoning.

METHODSSeventy eight rabbits were randomly divided into normal control group (N group, n=6), exposure groups (PQ group, n=24), hemoperfusion treatment group (HP treatment group, n= 24) and blank control group (HP group, n=24). The PQ, HPQ and HP groups were divided into 4 observation time groups (1, 3, 7 and 21 d). N group was exposed to 5 ml normal saline and PQ group was exposed to 50 mg/kg PQ by oral gavage. In 1 h after PQ exposure, HPQ group was exposed to the activated carbon hemoperfusion for 2 h. The content or activity of MDA, SOD and GSH-Px in lungs, livers and kidneys were detected, the expression levels of MMP-2, MMP-9 and TIMP-1 were measured with immunohistochemical SP method for all groups.

RESULTSThe contents of MDA in lungs, livers and kidneys of PQ and HPQ groups decreased and the activities of SOD and GSH-Px in lungs, livers and kidneys of PQ and HPQ groups increased with observation time. The expression levels of MMP-2, MMP-9 and TIMP-1 in PQ and HPQ groups enhanced on the first day, PQ group was most obvious. Along with the observation time extended, all kinds of positive expression were still high. Compared with normal control group, the activities of serum SOD and GSH-Px in PQ and HPQ groups declined significantly, but the contents of serum MDA increased; the expression levels of MMP-2, MMP-9 and TIMP-1 in lung, liver and kidney tissues increased obviously, the ration between MMP-9 and TIMP-1 significantly increased (P < 0.05). Compared with PQ group, the activities of SOD and GSH-Px in HPQ group significantly increased, the content of MDA declined, the expression levels of MMP-2, MMP-9 and TIMP-1 in lung, liver and kidney tissues declined obviously, the ration between MMP-9 and TIMP-1 significantly declined, but higher than N group, the differences were statistically significant (P < 0.05).

CONCLUSIONThe oxidative stress and MMPs may be involved in the pathogenesis of tissue injuries induced by paraquat. The treatment with HP could obviously reduce oxidative stress and the expression levels of MMP-2, MMP-9 and TIMP-1, enhance the ration between MMP-9 and TIMP-1. So HP treatment could play a role in rescuing the PQ poisoning and protecting the organs function.

Animals ; Female ; Hemoperfusion ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Matrix Metalloproteinases ; metabolism ; Oxidative Stress ; Paraquat ; poisoning ; Rabbits ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism

OBJECTIVETo explore the curative mechanism of acupuncture treatment on osteoarthritis (OA).

METHODSForty cases of female SD rats were randomly divided into a normal group, a model group, an acupuncture group and a medication group, 10 cases in each group. OA animal model was established by using the method of heel tendon resection for unilateral hind limb. The acupuncture group was treated with electroacupuncture at "Xiqian"(ST 35) and "Housanli"(ST 36), and the medication group with inunction of Diclofenac cream, and the normal group and the model group without any treatment. The expression of matrix metalloproteinase-1, 3 (MMP-1, MMP-3) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the cartilage were observed by immunohistochemistry.

RESULTSThere were significant differences among four groups. The expressions of MMP-1, MMP-3 and TIMP-1 in the model, acupuncture and medication groups were all significantly stronger than those in the normal group (all P < 0.01). The expressions of MMP-1 and MMP-3 in the acupuncture and medication groups were down regulated and TIMP-1 expression up-regulated with significant differences as compared with the model group (all P < 0.01), and the expressions of MMP-1 and MMP-3 in acupuncture group were significantly lower, while TIMP-1 expression significantly higher than that in the medication group (all P < 0.01).

CONCLUSIONAcupuncture can down-regulate the expression of MMP-1 and MMP-3 and up-regulate the expression of TIMP1, which is superior to that of Diclofenac cream, showing that acupuncture has a certain protective effect on cartilage from OA.

Acupuncture Therapy ; Animals ; Cartilage ; chemistry ; Female ; Immunohistochemistry ; Matrix Metalloproteinase 1 ; analysis ; Matrix Metalloproteinase 3 ; analysis ; Osteoarthritis, Knee ; metabolism ; therapy ; Random Allocation ; Rats ; Tissue Inhibitor of Metalloproteinase-1 ; analysis

AIMTo explore the possible impact of endogenous hydrogen sulfide (H2S) on the content and metabolism of collagen in rats with high pulmonary blood flow.

METHODSThirty-two male SD rats, weighing 120-140 g, were randomly divided into 4 groups (n = 8), shunt group, shunt + PPG (propargylglycine, an antagonist of endogenous H2S producing enzyme) group, sham group and sham + PPG group. Rats in shunt group and shunt + PPG group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of high pulmonary flow. In the sham group and sham + PPG group, rats experienced the same experimental processes except the shunting procedure. After 4 weeks of experiment, lung tissue H2S content of rat was determined by a modified sulfide electrode method. Pulmonary artery collagen I, collagen III, MMP-13 and TIMP-1 protein expressions of rat were investigated by immunohistochemistry.

RESULTSAfter 4 weeks of experiment, lung tissue H2S content increased significantly in rats of shunt group as compared with that of sham group (P < 0.05). Pulmonary artery collagen I and collagen III protein expression increased obviously in rats of shunt group as compared with that of sham group (P < 0.01). After administration of PPG for 4 weeks, lung tissue H2S content decreased significantly in rats of shunt + PPG group as compared with that of shunt group (P < 0.05). In contrast to rats in shunt group, collagen I and collagen III protein expression in pulmonary arteries of shunt + PPG group increased significantly, respectively (P < 0.05). Compared with rats of shunt group, pulmonary artery MMP-13, TIMP-1 and the ratio of MMP-13/TIMP-1 in shunt + PPG group down-regulated significantly (P < 0.05).

CONCLUSIONEndogenous H2S might play a protective regulatory role in the development of pulmonary hypertension and pulmonary vascular structural remodelling in rats by decreasing the content of pulmonary artery collagen resulting from catabolism of collagen.

Animals ; Collagen ; metabolism ; Hydrogen Sulfide ; metabolism ; Lung ; blood supply ; metabolism ; Male ; Matrix Metalloproteinase 13 ; metabolism ; Pulmonary Artery ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism