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Objective To observe the intervention effects of methylprednisolone (MPred) to acute lung injury(ALI) model of rats induced by oleic acid.Methods Thirty of Sprague-Dawley rats were randomly divided into three groups which were normal control group (NC,n =6),oleic acid model group (n =12),and MPred group (n =12).Rats were injected with oleic acid at dose of 0.1mg/ kg via caudal vein and then ALI model was established.The rats of NC group were injected with 0.1 mg/kg of normal saline instead of oleic acid.In NC group rats were sacrificed by blood collection at 6h after NS injection.Blood samples and tissues were collected and stored freezing.Samples of the other groups were collected at 2h and 6h after the last treatment.The observation indexes are histomorphology of lung tissue,the wet and dry weight of lung (W/D),index of quantitative assessment (IQA) score,partial pressure of oxygen in artery (PaO2),and SOD,MDA,MMP-9 and TIMP-1 levels in the blood plasma and lung tissue.Results Lung surface hyperemia relieved obviously and pink secretion from trachea of rats in MPred group decreased compared with oleic acid model group.In light microscope,compared with oleic acid model group,effusion of inflammatory cell in alveolar space of rats in MPred group eased.W/D of rats in oleic acid model group advanced obviously compared with that in NC group,W/D of rats in medrol group lowered obviously compared with that in oleic acid model group.IQA scores of rats in oleic acid model group advanced obviously compared with that in NC group,IQA score of rats in MPred group lowered obviously compared with that in oleic acid model group.PaO2 of rats in oleic acid model group lowered obviously compared with that in NC group,PaO2 of rats in MPred group advanced obviously compared with that in oleic acid model group.The level of SOD in plasma and lung tissue of rats in oleic acid model group lowered obviously compared with that in NC group,SOD level in plasma and lung tissue of rats in MPred group advanced obviously compared with that in oleic acid model group.The level of MDA in plasma and lung tissue of rats in oleic acid model group lowered obviously compared with that in NC group,MDA in plasma and lung tissue of rats in MPred group advanced obviously compared with that in oleic acid model group.Compared with the NC group,the level of MMP-9 in the plasma and lung tissue of oleic acid induced ALI rats increased and TIMP-1 levels decreased significantly.The level of MMP-9 in the plasma of rats decreased and TIMP-1 level increased significantly in MPred group at 2h and 6h.Conclusion MPred can improve lung pathological injury,increase PaO2 level,decrease lung W/D ratio and IQA scores by modulating the level or activity of the SOD and MDA and MMP-9 and TIMP-1 in the plasma and lung tissues.It is speculated that the effects of anti-inflammation and anti-exudation of sodium aescinate may due to affecting on oxidative stress response as well as the decomposition and remodeling of extracellular matrix during inflammatory lung injury of acute lung injury rats.
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Objective To explore the expression and significance of neutrophil gelatinase-associated lipocalin (NGAL) in lupus nephritis (LN) in mice. Methods Female MRL/lpr lupus mice (n=36) were randomly divided into the experimental group and intervention group, and female Kunming mice (n =18) served as controls. Each mice in the intervention group received intraperitoneal injection of 20 μg anti-mice interleukin (IL)-17 antibody. The serum concentrations of NGAL, IL-17,matrix metalloproteinase(MMP)-9 and tissue inhibitor of metalloproteinase (TIMP)-1 were measured by enzyme-linked immunosorbent assay (ELISA). And the protein expressions of NGAL, IL-17, MMP-9 and TIMP-1 in renal tissue were detected by immunohistochemisty. One-factor analysis of variance (ANOVA) or nonparametric rank sum test was used for the comparisons between the three groups. Associations between these factors were analyzed by Pearson′s test. Results The levels of serum NGAL, IL-17, MMP-9 and TIMP-1 in the experimental group were obviously increased as compared to those in the control group and intervention group [NGAL: (30.31±1.22) ng/ml vs (11.36±0.14) ng/ml, (20.09±0.35) ng/ml, F=986.524, P and the serum levels of IL-17, MMP-9 and MMP-9/TIMP-1(r=0.899, 0.789, 0.925, P<0.01). The protein expression of NGAL in renal tissue was positively correlated with IL-17, MMP-9 and MMP-9/TIMP-1 levels (r=0.929, 0.899, 0.723, P<0.01). Conclusion The level of NGAL in the serum and renal tissue is signifi-cantly increased in the MRL/lpr lupus mice. And it is closely correlated with the levels of IL-17 and MMP-9. Our results suggest the potential role of NGAL in the inflammation of lupus nephritis.
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Objective:To explore changes of peripheral blood levels of C reactive protein (CRP)and matrix metallo-proteinase-9 (MMP-9)in patients with coronary heart diseases (CHD)and percutaneous coronary intervention (PCI),and analyze their relationship with patients'prognosis.Methods:A total of 278 CHD patients undergoing PCI in our hospital from Jul 2009 to Apr 2011 were regarded as PCI group;another 234 CHD patients not receiving PCI were enrolled as CHD control group.According to results of coronary angiography,PCI group was further di-vided into single-vessel (n=143),double-vessel (n=92)and triple-vessel disease group (n=43).Changes of CRP and MMP-9 levels were compared between two groups in different time,and the relationship among these two in-flammatory factors and coronary disease,its prognosis was analyzed.Results:Compared with on admission,there were significant rise in levels of CRP [(2.43±0.62)mg/L vs.(2.87±0.73)mg/L,(2.98±0.87)mg/L]and MMP-9 [(12.63±2.68)ng/ml vs.(14.62±3.49)ng/ml,(19.62±4.63)ng/ml]in PCI group on 24h and 48h after PCI,P 0.05;The more severe coronary lesion was,the higher CRP and MMP-9 levels were,CRP and MMP-9 levels of triple-vessel group [(2.51 ±0.64)mg/L,(14.67±2.97)ng/ml]were significantly higher than those of single-vessel group [(1.83±0.51)mg/L,(9.68±1.42)ng/ml]and double-vessel group [(2.17±0.59) mg/L,(11.62±2.19)ng/ml],P <0.05~<0.01;incidence rates of cardiovascular events in patients with CRP≥3 mg/L and MMP-9≥15 ng/ml (33.3%,29.1%)were significantly higher than those of patients with CRP <3 mg/L and MMP-9<15 ng/ml (16.1%,18.2%)respectively,P <0.05 both.Conclusion:Serum levels of CRP and MMP-9 significantly rise in CHD patients on 48h after PCI,and their increasing level is related with extent of coronary ar-tery lesion and prognosis.
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Objective: To explore relationship among serum levels of homocysteine (Hcy), interleukin (IL)-10, matrix metalloproteinase (MMP)-9 and infarct volume in patients with acute cerebral infarction（CI）. Methods: A total of 125 acute CI patients were regard as acute CI group, and another 37 healthy subjects were enrolled as healthy control group. Enzyme linked immunosorbent assay (ELISA) method was used to measure serum levels of Hcy, IL-10 and MMP-9 of subjects; according to infarct volume, acute CI patients were divided into small infarction group (≤5cm3, n=47), medium infarction group (5~15 cm3, n=41) and large infarction group (≥15cm3, n=37); changes of levels of Hcy, IL-10 and MMP-9 were compared among all groups. Results: Compared with healthy control group, there were significant increase in levels of MMP-9 and Hcy, and significant decrease in IL-10 level in acute CI group. Compared with small and medium infarction groups, there was significant increase in serum Hcy level [(16.11±3.5) μmol/L, (16.79±3.4) μmol/L vs. (18.5±4.2) μmol/L, P<0.01, P<0.05] in large infarction group； Compared with small infarction group, there was significant increase in serum MMP-9 level [(161±27) ng/ml vs. (238±26) ng/ml, (256±35) ng/ml, P<0.05, P<0.01]；Compared with small infarction and medium infarction groups, there were significant decrease in IL-10 level [(0.60±0.17) ng/ml vs. (0.59±0.15) ng/ml, (0.31±0.12) ng/ml, P<0.01 both] in large infarction groups. Conclusion: Serum levels of homocysteine, interleukin-10 and matrix metalloproteinase-9 are related to onset of acute cerebral infarction, and may reflect risk degree of cerebral infarction.
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Objective To investigate the effects of the leflunomide active metabolite (A771726) on the expression of phorbol-12-myristate-13-acetate (PMA) -induced CD147, matrix metallo-proteinase (MMP)-2 and MMP-9 on THP-1 cells. Methods THP-1 cells were cultured in RPMI-1640 medium supplemented with 10％ fetal bovine serum. For all experiments, THP-1 cells were cultured at an initial density of 5×105/ml. Before A771726 treatment, cells were cultured with serum-free RPMI-1640 medium for 12 h, THP-1cells were co-cultured with PMA at three different concentrations of A771726 (5, 15 , 45 μg/ml) for 24 h.The mRNA expression of CD147, MMP-2 and MMP-9 was measured by real-time PCR. CD147 expression on the cells were evaluated by flow cytometric analysis. The activity of MMP-2 and MMP-9 were evaluated by gelatin zymography. Statistical differences among the groups were tested by one-way ANOVA or KruskalWallis test. Results The expression of CD147, MMP-2 and MMP-9 were upgraded by the PMA. The expression of CD147 on THP-1 cells was inhibited significantly by A771726 in a dose-dependent pattern (P＜0.01). The mean fluorescence intensity (MFI) of CD147 in positive control group was 109.5±3.8, the MFI in A771726 (5, 15, 45 μg/ml) group were 73.3±2.5, 64.5±2.3, 40.9±2.7, respectively. The expression of MMP-2, MMP-9 mRNA and the activity of MMP-2, MMP-9 in the supernatant was inhibited significantly by A771726 (P＜0.01). The expression of CD147 mRNA was not inhibited significantly by A771726 (P＞0.05).Conclusion Leflunomide active metabolite (A771726) can inhibit the expression of PMA-induced CD147,MMP-2 and MMP-9 on THP-1 Cells.
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Objective:To study the expression of extracellular matrix metalloproteinase (MMP-9) inducer CD147 and MMP-9 protein in hepatocellular carcinoma (HCC) and explore their correlation with invasion, metastasis, and prognosis of HCC.Methods:SP immunohistochemistry was used to evaluate the expression of CD147 and MMP-9 in 83 HCC tissues and 15 normal liver tissues. Results:The positive rate was 73.5% for MMP-9 and 68.7% for CD147, significantly higher than normal tissues (MMP-9: 6.7%;CD147: 0%, P<0.05). Expressions of CD147 and MMP-9 were positively correlated with the tumor size, pTNM stages, differentiation degree, the portal vein cancer embolism, and postoperative recurrence (P<0.05), but not with the patient's age, gender, the serum level of alpha-fetal protein (AFP), and the Child-Pugh grades. Patients with positive expression of CD147 and MMP-9 had shorter survival time than those with negative expression of CD147 and MMP-9. Expression of CD147 was positively related with expression of MMP-9 in HCC tissues.Conclusion:CD147 and MMP-9 had positive correlation with tumor size, pTNM stages, differentiation degree, portal vein cancer embolism, and postoperative recurrence. Detection of the expression of CD147 and MMP-9 may be helpful to make a comprehensive prediction of the prognosis of HCC.
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Objective:To investigate the role of serum inflammatory cytokines in the development of acute coronary syndrome (ACS). Methods: All of enrolled patients were diagnosed by clinical and coronary angiographic features and divided into four groups, the acute myocardial infarction (AMI) group, unstable angina pectoris (UAP) group, stable angina pectoris (SAP) group and control group. The values of high-sersitivity C-reactive protein(hs-CRP), matrix metallopeptidase 9(MMP-9) and tumor necrosis factor-a (TNF-a) in serum were measured by cytokine detection equipment system (B10-RAD Biological Technology Co.Ltd, USA) and analysed in four groups with statistics. Results: Compared with SAP and the control groups, the levels of TNF-a and MMP-9 were increased significantly in AMI group(P 0.05). It was found that there was positive relation between hs-CRP, MMP-9 and TNF-a by Pearson correlation analysis. Conclusion:There was obvious relation between coronary heart disease and inflammation. The cytokines characterized by the increases of hs-CRP, TNF-a and MMP-9 were involved in the formation and progression of atherosclerosis and served as markers of unstable plagues.
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Objective:To investigate the expression of human epidermal growth factor receptor 4 (HER4) and metastasis-related protein (MMP-9) in human esophageal carcinoma tissues, and their relationship with clinicopathological features of the disease. Methods:Immunohistochemical Envision technique was applied to detect the expressions of HER4 and MMP-9 in 45 specimens of esophageal carcinoma tissues, paracancerous tissues and normal tissues. Statistical method was used to analyze the association of the positivity of HER4 with clinical pathological index and MMP-9 expression.Results:The positive rates of HER4 expression were 73.3%, 33.3%, and 2.2% in 45 specimens of esophageal carcinoma tissues, paracancerous tissues, and normal tissues, respectively. The expression of HER4 was correlated with TNM stage and lymph node metastasis (P0.05). The expression of MMP-9 correlated with T stage, invasion depth, TNM stage, and lymph node metastasis (P<0.05). Conclusion:The expression of HER4 is apparently different in esophageal carcinoma tissues, paracancerous tissues, and normal tissues. Its positive expression in esophageal carcinoma tissues is correlated with TNM stage and lymph node metastasis. The expression of MMP-9 in esophageal carcinoma tissues is correlated with the T stage, TNM staging, and lymph node metastasis. The positive expression of HER4 in esophageal carcinoma tissues is associated with the expressions of MMP-9.
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Objective To investigate the relationship between acute cerebral infarction(CI)and matrix metalloproteinase-9(MMP-9)serum level and polymorphism(C-1562T)in Han population.Methods One hundred and one patients with acute CI from the department of neurology of Taizhou Hospital were included and 114 healthy persons were selected from physical examination as the control group.Serum MMP-9 level was measured by enzyme-linked immunosorbent assay(ELISA).At the same time.genotype was determined by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)for the common C-1562T functional promoter polymorphism of the MMP-9 gene.The serum MMP-9 level and genotype frequencies of the MMP-9 gene between the patients and the control group were analyzed.Results The genotype frequencies of the MMP-9 gene C-1562T polymorphism of the two groups were in Hardy-Weinberg equilibrium.The results of individual polymorphisms analysis showed that the frequencies of CT and TT genotypes in the C-1562T polyporphismin were of no significant difference between the patients group with CI(13.9%)and the control group(13.2%).The frequencies of-1562T allele was of no statistical difference between the CI group(6.9%)and the control group(7.5%).But serum levels of MMP-9 in CI patients group((138.9±121.8)ng/ml)were significantly higher than in the control group((18.4±4.6)ng/ml,t=9.93,P=0.00).Conclusions Serum level of MMP-9 obviously is increased after ischemic stroke in 48 hours.But genetic polymorphism in MMP-9 promoter(C-1562T)has no definite relationship with MMP-9 genetic expression and CI in the Han population of China.Therefore,the relationship between genetic polymorphism in MMP-9 promoter(C-1562T)and ischemic stroke needs further investigation.
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Objective To investigate the expression of MMP-9, p53 protein in primary hepatocarcinoma (PHC) and the relationship between the clinicpathological in PHC. Methods The expression of MMP-9 and p53 in 74 eases with PHC and microvessel density(MVD) were detected by immunohistochemistry,the clinical pathological parameters were also analyzed. Results In hepato-cellular carcinoma, the expression of MMP-9 and p53 were closely related to MVD, tumor size, aggressive behavior(vascular metastasis and tumor envelope) ( P < 0.05 ). The positive correlation was found between the expression of two factors in PHC. Conclusion MMP-9 and p53 might be impor-tant markers in the carcinogenesis,progression and prognosis of PHC.
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Objective To study the accuracy of plasma matrix metalloproteinase-9 (MMP-9) predicting hemorrhagic transformation (HT) in ischemic stroke. Methods The studies on plasma MMP-9 predicting HT in ischemic stroke were included, in which diagnostic criteria, methods, and subjects were clearly described, regardless of language of publication and type of studies. MEDLINE (Apr, 1966 to 2006), EMBase (Apt, 1966 to 2006), CNKI (Feb, 1977 to 2006), and wanfangshuju (1989 to 2005) were searched and the references lists of eligible studies were manually searched. Two reviewers independently evaluated the quality of studies and extracted data. The data were analyzed using the Revman 4. 2 or SPSS 11.0. Results Five trials including 497 patients fulfilled the inclusion criteria, 109 of the 497 patients developed HT. The heterogeneity among studies was great (xspecialty 89. 9% (95% CI: 86. 4%-93.5%), sensitivity 86. 5% (95% CI: 77. 3%-96. 8%). Conclusions The values of plasma MMP-9 may be an independent predictor of hemorrhagic transformation. The number of patients in the included studies is not large enough, and the results need to be confirmed in further studies.
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Objective To approach the effects of MMP-9 antisensc oligonucleotide(ASODN)on human lung adcnocarcinoma(A549) cell apoptosis and proliferation capability.Methods MTT.was used to analyze the effect that MMP-9 transfection on A549 cell growth.Flow cytometry was used to analyze the ratio of cell proliferation and apoptosis.RT-PCR WaS used to detect the expression of MMP-9mRNA and Western blot was used to detect the changes of MMP-9 protein.Results In some degree,MMP-9 ASODN that inhibited the survival rate of the A549 cell presented concentration and time dependence manner.The best dependence concentration of ASODN was 600nmol/L for 24 hour.After trasnfection of MMP-9ASODN to A549 cell.the perceive of A549 apoptosis cell was significandy higher than that in control group(P＜0.01).When the antisense oligonucleotides concentration is 600nmol/L and the action time is 48 hours,the relative expression level of MMP-9mRNA and MMP-9 protein are obviously less than that in control group(P＜0.01).Conclusion MMP-9ASODN may down regulate the expression of MMP-9mRNA and MMP-9 protein,effectively inhibit the proliferation of A549 cell and promote the apoptosis of A549 cell.
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Objective: To investigate the expressions of matrix metalloproteinase-9 (MMP-9) and E-cadherin protein in human colorectal carcinoma and their correlation with tumor infiltration and metastasis as well as evaluate the relationship between the expressions of the two proteins. Methods: The expressions of MMP-9 and E-cadherin protein were detected by immunohistochemical staining (streptavidine-peroxidase method) in 52 cases of colorectal carcinoma (including 3 cases of colorectal adenomas with malignant change). Ten cases of normal colorectal tissues were as control. Results: (1) The over-expression of MMP-9 was closely related with lymph node metastasis and invasion of colorectal carcinoma. The positive rate of MMP-9 expression in the patients with lymph node metastasis (100%) was significantly higher than those without lymph node metastasis (89%). The positive expression of MMP-9 was significantly higher in patients who survived less than 5 years than those who survived more than 5 years (P 0.05). Conclusion: Over-expression of MMP-9 and reduction or loss of E-cadherin expression has significant correlation with metastasis and infiltration of colorectal carcinoma. The positive expression of MMP-9 closely correlated with the abnormal expression of E-cadherin.
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0.05).However the MMP-2 and pro MMP-2 concentrations were still higher 30 d after implantation(13 935?2 167,15 628?1 739 INT?mm~2 vs 11 568?2 219,13 649?1 894 INT?mm~2,P
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Objective To investigate the correlations of serum matrix metalloproteinase-2, -9 (MMP-2, MMP-9) contents and tissue expressions in hepatocellular carcinoma with tumor invasion and metastasis. Methods Serum MMP-2, MMP-9 contents were detected in 40 patient with hepatocellular carcinoma and 20 healthy controls by ELISA; the expressions and distributions of MMP-2 and MMP-9 in 40 patients and 10 normal tissues were detected by immunohistochemical method. Results Serum MMP-2, MMP-9 contents were significantly elevated in cancer samples compared with normal serum (P
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OBJECTIVE To construct a lentiviral vector of RNA interference (RNAi) of MMP-9 gene and observe its inhibitive role on the invasion of laryngeal cancer cells.METHODS The effective sequence of siRNA targeting MMP-9 gene was confirmed.Both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pLVTHM vector,which contained H1 promoter and green fluorescent protein (GFP).The resulting lentiviral vector containing MMP-9 shRNA was called LV2 shMMP-9,and it was confirmed by PCR and sequencing.After that,MMP-9 shRNA was transfected into Hep-2 cells and western blot was used to test the expression of MMP-9.At last,Boyden Chamber was used to observe the invasion of the Hep-2 cells. RESULTS PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of MMP-9 (LV2shMMP-9) producing MMP-9 shRNA was constructed successfully. The titer of concentrated virus was 8?1010TU /L. Western blot showed that the expression of MMP-9 was negative in the MMP-9 siRNA transfected Hep-2 cells. And Boyden Chamber showed the invasive capability of Hep-2 cells transfected MMP-9 siRNA were obviously decreased.CONCLUSION The lentivirus RNAi vector of MMP-9 was constructed successfully,and MMP-9 silence can inhibit invasion of laryngeal cancer in vitro.
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Heat shock protein 70 (Hsp70) release and its effects on pro-inflammatory cytokine production have been controversial. In this study, we investigated whether Hsp70 could be released from monocytes and activates matrix metalloproteinase-9 (MMP-9) gene expression. Hsp70 overexpression in human monocytic cell line U937 was found to increase PMA- induced MMP-9 expression and enhance cell motility. Hsp70 cDNA transfectants released Hsp70 protein into culture supernatants, and a part of released Hsp70 subsequently was bound to the surface of U937 cells. Addition of culture medium containing the extracelluar Hsp70 led to an increase not only in proMMP-9 secretion, but also the invasiveness of U937 cells through Matrigel or human umbilical vascular endothelial cells (HUVEC) in vitro. Immunodepletion of Hsp70 abolished its effect on MMP-9 expression. The released Hsp70 activated nuclear factor kappa B (NF-kappa B) and activating protein-1 (AP-1), which led to the activation of MMP-9 transcription. Taken together, these results suggest that extracellular Hsp70 induces the expression of MMP-9 gene through activation of NF-kappa B and AP-1.
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Humans , U937 Cells , Transfection , Transcription Factor AP-1/metabolism , NF-kappa B/metabolism , Matrix Metalloproteinase 9/metabolism , HSP70 Heat-Shock Proteins/metabolism , Gene Expression Regulation , Culture Media, Conditioned/pharmacology , Cell Movement/drug effectsABSTRACT
Objective To determine the expression of Vascular Endothelial Growth Factor(VEGF) and Matrix Metalloproteinases-9(MMP-9) in non-small cell lung cancer(NSCLC) specimens and try to find out the possible roles of VEGF and MMP-9 in the invasion and metastasis of NSCLC.Methods Immunohistochemical SABC method was used to detect the expression of VEGF and MMP-9 in 23 non-cancerous pulmonary diseases tissues,30 normal lung tissues near the NSCLC and 111 NSCLC tissues.Results The expression levels of VEGF and MMP-9 in the NSCLC tissues were significantly higher than those of the control groups(P
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Objective Research the correlation between expression of MMP-2 in colorectal cancer and patho- logical factors,or biological behaviors in colorectal cancer.Methods The expression of MMP-2 in 68 cases with col- orectal cancer were detected by immunohistochemical staining.Results The expression of MMP-2 in colorectal can- cer was not correlated with tumor region and histologic type,but related with depth of tumor invasion and metasta- sis.The frequency of positive cases in patients with colorectal cancer with of lymph,node metastasis was significantly higher than that in patients without lymph node metastasis(P
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Objective To investigate the therapeutic effect of buflomedil hydiochoride for on jection inpro- gressive ischemic stroke.Methods 60 patients with cerebral infarction were divided into buflomedil hydrochoride for injection treatment group and control treatment.Then the nepal function deficits of patients were evaluated before and after 15 days.Serum MMP-9 levels in vein blood samples of ischemie stroke were detected.Results The inci- dence rate of progressive ischemic stroke in treatment group of buflomedil hydrochoride for injection was lower than that of control treatment group(P