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 observe the effects of Shuxuening Injection (SI) on the levels of serum matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase (TIMP-1) in acute exacerbated chronic obstructive pulmonary disease (COPD) patients.

METHODSSeventy-nine patients with acute exacerbated COPD were randomly assigned to the treatment group (39 cases) and the control group (40 cases). Routine therapies for COPD were given to patients in the control group, while 15 mL SI was given to those in the treatment group by intravenous dripping, twice daily for total 14 days. The forced expiratory volume in one second (FEV1) and forced vital capacity (FVC) were detected by Spirometer. The FEV1/FVC (%) and the FEV1% were calculated. The levels of serum MMP-9 and TIMP-1 were detected using ELISA before and after treatment, and compared with 20 healthy subjects as the control.

RESULTSThe FEV1, FVC, FEV1/FVC (%), and FEV1% were significantly improved after treatment in the treatment group when compared with before treatment and with the control group (P < 0.05, P < 0.01). When compared with before treatment and with the control group, the levels of serum MMP-9, TIMP-1, and the ratio of MMP-9/TIMP-1 decreased more significantly in the treatment group after treatment (P < 0.01). Correlation analyses showed that obvious negative correlation existed between the levels of serum MMP-9 and TIMP-1 and FEV1/FVC (%) (r = -0.677, -0.629, P < 0.01). Obvious negative correlation existed between the levels of serum MMP-9 and TIMP-1 and FEV1% (r = -0.562, -0.661, P < 0.01). Furthermore, obvious negative correlation also existed between the ratio of MMP-9/ TIMP-1 and FEV1%, as well as FEV1/FVC (%) (r = -0.732, -0.891, P < 0.01).

CONCLUSIONSSI could improve the pulmonary ventilation function of acute exacerbated COPD patients. One of its mechanisms might be correlated with lowering the serum levels of MMP-9 and TIMP-1, and correcting the imbalance of MMP-9/TIMP-1.

Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Humans ; Male ; Matrix Metalloproteinase 9 ; blood ; Middle Aged ; Phytotherapy ; Pulmonary Disease, Chronic Obstructive ; blood ; drug therapy ; Tissue Inhibitor of Metalloproteinase-1 ; blood

OBJECTIVE: To study the effect of quercetin, an inhibitor of matrix metalloproteinases 9 (MMP-9), on the growth and metastasis of lung cancer cells and the underlying mechanisms.
 METHODS: We evaluated the inhibitory effect and the inhibitory kinetics of quercetin on MMP-9 by ELISA and enzyme inhibition kinetics, and the inhibitory effect of quercetin on the growth of lung cancer cell (A549) by MTT. The effect of quercetin on levels of MMP-9 (mRNA and protein) and TGF-β1 (protein) in A549 were measured by RT-PCR and Western blot, respectively. The synergistic inhibition effect of quercetin plus TIMP-1 on the growth of lung cancer cell A549 was discussed.
 RESULTS: Quercetin induced the apoptosis of A549. It was a reversible competitive inhibitor of MMP-9 (half inhibition rate IC50 of 5.25 μmol/L, inhibition constant Ki was 2.18 μmol/L). With the increase in quercetin concentration, the levels of MMP-9 (mRNA and protein) and TGF-β1 (protein) were decreased, and the number of tumor cells on wear filter membrane was reduced. The combination of quercetin (at low concentrations) with TIMP-1 showed synergistic inhibitory effect on the growth of A549 cells. 
 CONCLUSION Quercetin is a competitive inhibitor of MMP-9 and could downregulate the expression of MMP-9 and TGF-β1, which plays an important role in A549 apoptosis.
Apoptosis ; Cell Line, Tumor ; drug effects ; Down-Regulation ; Humans ; Lung Neoplasms ; pathology ; Matrix Metalloproteinase 9 ; metabolism ; Matrix Metalloproteinase Inhibitors ; pharmacology ; Neoplasm Metastasis ; Quercetin ; pharmacology ; RNA, Messenger ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; pharmacology ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo compare the effects of matrix metalloproteinase (MMP) inhibitor doxycycline, losartan, and their combination on the expression of MMP-8, 13, tissue inhibitor of MMP-1, 2 (TIMP-1, 2), and collagen remodeling in the noninfarcted myocardium after acute myocardial infarction (AMI) in rats.

METHODSTwo hundred and fifty-four AMI rats, induced by left coronary ligation, were randomly assigned to the following groups: (1) AMI controls group (n = 64); (2) doxycycline group (30 mg x kg(-1) x d(-1), n = 63); (3) losartan group (10 mg x kg(-1) x d(-1), n = 62); (4) concomitant doxycycline and losartan group (30 and 10 mg x kg(-1) x d(-1) respectively, n = 65); and (5) Sham-operated rats (n = 30), which were randomly selected to serve as noninfarction controls. Each group was further divided into three subgroups of 1, 2, and 4 weeks that received treatment. After the completion of treatment, the rats were killed. The mRNA and protein expression of MMPs and TIMPs in the noninfarcted myocardium were quantified by RT-PCR and Western blot, respectively. The type I and type III collagen volume fraction (CVF) of the noninfarced myocardium were assessed immunohistochemically.

RESULTSNo significant difference existed in myocardial infarction sizes among the 12 subgroups of AMI controls and the three treatment groups (42%-48%, all P > 0.05). Compared with sham operated rats, the mRNA and protein expression of MMP-8 and 13 significantly increased by 39%-183% in all three subgroups of AMI controls (all P < 0.05), except both of their mRNA expressions in 2-week subgroups; the mRNA and protein levels of TIMP-1 increased only in 1-week subgroup of AMI controls by 104% and 67%, respectively (both P < 0.05); the mRNA of TIMP-2 increased in all 1, 2, and 4-week subgroups by 144%-232% (all P < 0.05), but its protein expression lagged and only enhanced in 2 and 4-week subgroups of AMI controls by 231% and 332%, respectively (both P < 0.05). Meanwhile, both type I and type III CVF of noninfarcted myocardium significantly increased in all three subgroups of AMI controls (type I CVF: 3.01%-5.64% vs 1.53%-1.67%, P < 0.01-0.001; type III CVF: 2.19%-4.42% vs 1.46%-1.59%, P < 0.05-0.001), with type I CVF being higher in 4-week than in 1 and 2-week subgroups (5.64% vs 3.01% and 3.02% respectively, all P < 0.05). Compared with AMI controls, all three kinds of treatment significantly reduced the increased mRNA and protein expressions of MMP-8, 13 and TIMP-1, 2 after AMI by 14%-60% (all P < 0.05), as well as type I/III CVF in their 2 and 4-week subgroups (type I CVF: 1.56%-2.38% vs 3.02%-5.64%, P < 0.05-0.001; type III CVF: 1.92%-2.65% vs 4.19%-4.42%, P < 0.05-0.01), except for doxycycline's effect on type III CVF in any of its three subgroups (all P > 0.05). Among the three treatment groups, significant differences existed in the above mentioned indicators only at some subgroup levels (all P < 0.05).

CONCLUSIONSLike losartan, doxycycline can also suppress the enhanced mRNA and protein expression of MMP-8, 13 and TIMP-1, 2, and reduce type I collagen deposition in the noninfarcted myocardium after AMI in rats. However, it has no effect on type III collagen deposition.

Angiotensin II Type 1 Receptor Blockers ; pharmacology ; Animals ; Collagen Type I ; biosynthesis ; genetics ; Collagenases ; biosynthesis ; genetics ; Doxycycline ; pharmacology ; Drug Synergism ; Female ; Losartan ; pharmacology ; Matrix Metalloproteinase 13 ; Matrix Metalloproteinase 8 ; biosynthesis ; genetics ; Matrix Metalloproteinase Inhibitors ; Myocardial Infarction ; metabolism ; Myocardium ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics ; Tissue Inhibitor of Metalloproteinase-2 ; biosynthesis ; genetics ; Tissue Inhibitor of Metalloproteinases ; biosynthesis ; genetics

OBJECTIVETo investigate the molecular mechanism of dermal damage in heat shock-induced skin aging by observing the expressions of metalloproteinase-1 (MMP-1) and tissue inhibitor of MMP-1 (TIMP-1) in retinoic acid-treated cultured human fibroblasts with heat shock.

METHODSCultured human fibroblasts were treated with tazarotene or all-trans-retinioic acid (at-RA) after heat shock for 30 min in 43 degrees celsius; water bath. Twenty-four hours later, MMP-1 and TIMP-1 contents in the supernatant of the cell culture medium were measured using enzyme-linked immunosorbent assay (ELISA).

RESULTSBoth tazarotene and at-RA dose-dependently reduced the expression of MMP-1 and increased the expression of TIMP-1 in cultured human fibroblasts exposed to heat shock, and tazarotene produced stronger effect than at-RA.

CONCLUSIONRetinoic acid can reduce the expression of MMP-1 and increase the expression of TIMP-1 in cultured human fibroblasts, suggesting its therapeutic potential for heat shock-induced skin aging.

Cells, Cultured ; Fibroblasts ; cytology ; metabolism ; Heat-Shock Response ; Humans ; Matrix Metalloproteinase 1 ; genetics ; metabolism ; Nicotinic Acids ; pharmacology ; Skin Aging ; radiation effects ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism ; Tretinoin ; pharmacology

OBJECTIVETo explore the roles of matrix metalloproteinase-1(MMP-1) and tissue inhibitor of metalloproteinase-1(TIMP-1) in the pathogenesis of endometriosis and the effects of estrogen and progestin on their expression.

METHODSImmunohistochemistry and RT-PCR were employed to detect the expression of MMP-1 and TIMP-1 in the ectopic tissues of 35 patients with endometriosis, 22 eutopic endometrium tissues from women with endometriosis and 28 normal controls. Fifty-nine nude mice were injected with human late secretory endometrial chippings and randomized into estrogen group, progestin group, estrogen-progestin group and control group with corresponding treatments. The implantation rates and graft morphology were observed and MMP-1 and TIMP-1 expressions in the grafts detected by immunohistochemistry.

RESULTSTypical endometrial glands and stroma were observed in all the groups with comparable implantation rates. The administration of progestin was associated with multiple peritoneal implantation sites and significantly larger implants. The transplanted endometria showed proliferative or secretory changes with estrogen or progestin administration. MMP-1 expression significantly increased and TIMP-1 expression decreased with increased MMP-1/TIMP-1 ratio in human and nude mouse ectopic endometria in comparison with those in normal endometria (P<0.05, P<0.01). MMP-1 expression was higher in estrogen and estrogen-progestin groups than in the control group, and was lower in the 3 sexual hormone-treated groups than in the control group. MMP-1 mRNA expression in the eutopic endometrium was significantly higher than that in the normal endometria.

CONCLUSIONProgestrin can not inhibit MMP-1 expression or the effect of estrogen on ectopic endometrium known as progestin resistance. The high expression of MMP-1 and low expression of TIMP-1 in endometriotic tissues confer strong invasiveness of ectopic endometrial tissue, especially in eutopic endometrial tissue, and may play an important role in the pathogenesis of endometriosis.

Adult ; Animals ; Endometriosis ; metabolism ; Estrogens ; pharmacology ; Female ; Humans ; Matrix Metalloproteinase 1 ; genetics ; metabolism ; Mice ; Mice, Nude ; Middle Aged ; Progestins ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism

OBJECTIVE: To explore the relationship between protein kinase C (PKC) and matrix metalloproteinase (MMPs)/tissue inhibitor of metalloproteinase (TIMPs) in human mesangial cells under the high glucose medium, and to analyze the effect of PKC and MMPs/TIMPs in diabetes nephropathy (DN). METHODS: Normal human mesangial cells (NHMC) were divided into 4 groups: a control group(N, 5 mmol/L glucose), a high glucose group (H, 30 mmol/L glucose), a PKC inhibition group (P, 30 mmol/L glucose plus 10-5 mol/L chelerythrine chloride), and an mannitol group (M, 5 mmol/L glucose plus 25 mmol/L mannitol). Cell proliferation was measured by MTT at 24,48 or 72 hours. The activity of PKC was measured by ELISA and the mRNA and protein expressions of MMP2, 9 and TIMP1, 2 were examined by RT-PCR and Western blot. RESULTS: High glucose increased the activity of PKC as well as the expressions of mRNA and protein of MMP2, 9 and TIMP1, 2.The ratio of MMP-2/TIMP-2 and MMP-9/TIMP-1 was significantly decreased in the high glucose group compared with that of the control group (P<0.05). The mRNA and protein expressions of MMP2, 9 and TIMP1 were significantly increased in the PKC inhibition group compared with the control group (P<0.01). The ratio of MMP-2/TIMP-2 and MMP-9/TIMP-1 increased in the inhibition group compared with that of the high glucose group (P<0.05 or P<0.01). The activity of PKC was negatively correlated with the protein ratio of MMP-2/TIMP-2 and MMP-9/TIMP-1(rý-0.651,rý-0.702, both P<0.05). CONCLUSION High glucose can activate PKC in mesangial cells. The activity of PKC influences the expression of MMPs/TIMPs in the progressing of DN.
Cells, Cultured ; Glucose ; adverse effects ; pharmacology ; Humans ; Hyperglycemia ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mesangial Cells ; metabolism ; Protein Kinase C ; metabolism ; Protein Kinase Inhibitors ; pharmacology ; RNA, Messenger ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism

OBJECTIVETo investigate the effect of the active components from Red Paeonia and Rhizoma chuanxiong (in Xiongshao Capsule, XSC) on matrix metalloproteinases (MMPs) in rabbit model of atherosclerosis.

METHODSFifty New Zealand rabbits were randomly divided into the normal control group (A), the model control group (B), the Simvastatin treated group (C), the low-dose XSC treated group (D) and the high-dose XSC treated group (E), 10 in each group. Rabbits in the normal control group were fed with normal diet, while those in the other four groups were fed with high fat diet and duplicated after two weeks feeding into model of abdominal aortic atherosclerosis by balloon angioplasty. In the 6 successive weeks feeding of high fat diet, Simvastatin 2.5 mg/kg, XSC 0.24 g/kg and 0.48 g/kg per day was given respectively to the rabbits in the three treated groups. Blood sample was collected for determining the level of blood lipids; serum MMP-3 and MMP-9, and tissue inhibitor of metalloproteinase-1 (TIMP-1) with enzyme-linked immunoassay; and the protein expression of MMP-3 and cluster of differentiation antigen 40 ligand (CD40L) in plaque were detected with immunohistochemical method.

RESULTSCompared with Group B, the serum levels of MMP-3 and MMP-9; the expression of MMP-3 and CD40L in plaque; and the blood content of total cholesterol in the three treated groups were significantly lower (P < 0.05 or P < 0.01). Besides, the content of triglyceride and low-density lipoprotein cholesterol were significantly reduced in Group C, while the TIMP-1 showed no statistical difference among different groups (P > 0.05).

CONCLUSIONThe active components of Red Paeonia and Rhizoma chuanxiong play a definite role in stabilizing the atherosclerotic plaque in rabbits, one of their possible mechanisms may be by way of inhibiting the expressions of MMP-3, MMP-9 in vascular walls and blood serum.

Animals ; Atherosclerosis ; drug therapy ; enzymology ; Drugs, Chinese Herbal ; pharmacology ; Male ; Matrix Metalloproteinase 3 ; blood ; Matrix Metalloproteinase 9 ; blood ; Paeonia ; chemistry ; Phytotherapy ; Rabbits ; Random Allocation ; Tissue Inhibitor of Metalloproteinase-1 ; blood

OBJECTIVETo observe the effect of salviandic acid B (SA-B) on MMP-2/9 and TIMP-2 of fibrotic cardiac tissues in rats and explore the action mechanism of SA-B anti-fibrosis of heart.

METHODVentricular remodeling model was induced by abdominal aortic banding (AAB) in rats. Rats were randomly divided into 6 groups: normal, model, SA-B high, SA-B middle, SA-B low and captopril control group. Histological changes of heart were observed with hemotoxylin and eosin (H&E) staining and Sirius red staining. Hydroxyproline (Hyp) content in heart tissue was measured by hydrolysis method. Expression of heart tissue collagen NIV, MMP-2/9 and TIMP-2 were analyzed with Western blot The activities of heart tissue MMP-2 were determined with gelatin zymography substrate degradation method.

RESULTSA-B treated groups had lower heart inflammation and lower heart Hyp content; decreased Collagen deposit and alleviated cardiac fibrosis. SA-B treated groups obviously decreased the expression of Collagen IV, MMP-2/9 and TIMP-2. The activity of MMP-2 was decreased in treated SA-B treated groups.

CONCLUSIONThe mechanism of SA-B action against cardiac fibrosis may be related to down-regulating the expression of TIMP -2 and the activity of MMP-2/9, thus protect the normal basal membrane.

Animals ; Basement Membrane ; drug effects ; enzymology ; Cardiomegaly ; drug therapy ; enzymology ; genetics ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism

OBJECTIVETo observe the in vitro effect of compound 861 (Cpd 861) on tissue inhibitor of metalloprotenase 1 (TIMP1) mRNA levels of HSC-T6 cell.

METHODSHSC-T6 cells were exposed in different concentrations of Cpd 861 (0.25~1.0 mg/ml) for 48 hours. The TIMP1 level was measured by the quantitative reverse-transcription polymerase chain reaction (RT-PCR).

RESULTSThe TIMP1 mRNA levels of HSC-T6 cells at different concentrations of Cpd 861 were lower (2.50 0.71, 0.50 0.01, 0.11 0.03) than those of the normal control (3.78 0.67, P<0.05 or P<0.01).

CONCLUSIONSThe antifibrotic mechanism of Cpd 861 is partly due to its downregulation on TIMP1 mRNA levels of HSC-T6 cells.

Animals ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression ; drug effects ; RNA, Messenger ; biosynthesis ; drug effects ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics