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Objective:To investigate the role and underlying mechanisms of inhibiting high mobility group box-1 (HMGB1) in the expression of matrix metalloproteinase-9 (MMP-9) in spinal cord astrocytes (AS) in rats after spinal cord injury (SCI).Methods:After an SCI model was established in Sprague-Dawley (SD) rats using a modified Allen's Weight-Dropping method and ethyl pyruvate (EP) or glycyrrhizin (GL) was used to inhibit the effect of HMGB1, the rats were divided into a sham group, an SCI group, an SCI+EP (50 mg/kg) group, and an SCI+GL (100 mg/kg) group. The expression levels of glial fibrillary acid protein (GFAP) and MMP-9 in spinal cord AS were observed. After the spinal cord AS in SD rats was cultured and incubated by the oxygen-glucose deprivation/reoxygenation (OGD/R) procedure, the expression of MMP-9 protein was detected at 6 h/R 6 h, 12 h, 24 h, and 48 h after OGD. The time point with the highest expression was chosen in the subsequent experiments as an OGD/R group. HMGB1 was inhibited by HMGB1 shRNA or EP to observe the effect of HMGB1 on the expression of MMP-9 protein in AS treated with OGD/R. Then, toll-like receptor 4 (TLR4) inhibitor, TIR-domain-containing adaptor inducing interferon- β (TRIF) inhibitor, and nuclear factor-kappa B (NF- κB) inhibitor were used to investigate the effects of TLR4/TRIF/NF- κB signaling pathway during the regulation of HMGB1 on MMP-9 in vitro. Results:Western blot showed that the expression of MMP-9 protein in the spinal cord was significantly increased in rats at 1 d after SCI, and the expression of MMP-9 protein in the SCI+EP group and the SCI+GL group was significantly lower than that in the SCI group ( P<0.001). Immunofluorescence showed that GFAP and MMP-9 proteins were co-localized in the spinal cord after SCI, and the expression of GFAP and MMP-9 proteins in the SCI+EP and SCI+GL groups was significantly lower than that in the SCI group ( P<0.05). Since the expression of MMP-9 protein in the spinal cord AS cultured in vitro was significantly higher in the OGD 6h/R 12h group than that in the normal group and the OGD 6h/R 6h, 24, and 48 h groups, the OGD 6h/R 12h was taken as the OGD/R group. The MMP-9 protein expression in AS in the OGD/R+HMGB1 shRNA group and the OGD/R+EP group was significantly lower than that in the OGD/R group ( P<0.001). In the cultured AS, moreover, inhibiting TLR4, TRIF, and NF- κB reduced MMP-9 protein expression after OGD 6 h/R 12 h when compared with that in the OGD/R group ( P<0.001). Conclusions:HMGB1 inhibition may result in a reduction in MMP-9 expression both in the spinal cord AS in SCI rats and in AS after OGD/R treatment in vitro. HMGB1 may regulate MMP-9 protein expression in AS after OGD/R treatment via the TLR4/TRIF/NF- κB signal pathway.
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Objective:To investigate the level and significance of CD64 index, matrix metalloproteinase-9 (MMP-9) and serum amyloid A (SAA) in peripheral blood of patients with severe carbapenem resistant Enterobacteriaceae (CRE) infection.Methods:A total of 61 patients with severe CRE infection who were admitted to the neurosurgery department of Kashgar First People′s Hospital from January 2019 to January 2022 were selected as the CRE group, and 100 patients with severe carbapenem sensitive Enterobacteriaceae (CSE) infection were selected as the CSE group. The difference in clinical data between the two groups was compared, and the difference in clinical data between the dead and surviving patients in the CRE group was compared. The value of CD64 index, MMP-9 and SAA in differential diagnosis of CRE was analyzed. Logistic regression was used to analyze the influencing factors of prognosis in patients with CRE infection.Results:The age, hypertension, lung disease, liver and kidney disease, comorbidities≥2, antibiotic use≥2 combinations, antibiotic use time>10 days, proportion of carbapenem use, CD64 index, MMP-9, and SAA of the CRE group patients were significantly higher than those of the CSE group patients (all P<0.05). The area under the receiver operating characteristic (ROC) curve for CD64 index, MMP-9, and SAA differential diagnosis of CRE was 0.857, 0.701, and 0.655, respectively (all P<0.05). In the CRE group, the age , the score of Acute Physiological and Chronic Health Status Ⅱ (APACHE Ⅱ) score at admission, diabetes, liver and kidney diseases, comorbidities≥2, the proportion of carbapenems, CD64 index, MMP-9 and SAA of dead patients were significantly higher than those of survivors (all P<0.05). Logistic regression analysis showed that age, APACHE Ⅱ score at admission, comorbidities≥2, CD64 index, MMP-9, and SAA were influencing factors for the prognosis of severe CRE patients (all P<0.05). Conclusions:The peripheral blood CD64 index, MMP-9, and SAA have certain application value in the diagnosis of neurological severe CRE infection, and are also influencing factors for the prognosis of CRE infected patients.
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@#Introduction: Smoking causes cardiovascular risk which may alter the stability between the production and degradation of the extracellular matrix. Matrix metalloproteinase-9 (MMP-9) is a zinc-containing endopeptidase that degrades the extracellular matrix and plays a vital role in tissue remodeling. As a result, elevated serum MMP-9 levels produced by smoking, particularly at young age, raise the risk of future CHD. So this study aims to find out the possible relationship between circulating MMP-9 and the risk of cardiovascular disease in young smokers. Methods: The study was conducted on smokers with CHD subjects attending cardiology and medicine OP of the SRM Medical College Hospital and research center Tamil Nadu, India. The study group was divided into three groups. Group 1 includes 120 healthy controls as nonsmokers, Group 2 includes 120 smokers with Coronary heart disease (CHD), and Group 3 includes 120 smokers with diabetes and CHD subjects in the age group of 20-55 years. Serum MMP-9, hs-CRP, and APO-E levels were measured using the ELISA method and the lipid level was measured enzymatically using AU480 automatic analyzer (back man coulter). Results: The mean serum MMP-9, hs-CRP, and APO-E levels were significantly higher in both groups (p<0.05) when compared to controls. The study also shows a significant positive association between MMP-9 with hs-CRP, APO-E, smoking burden, and smoking intensity. Conclusion: The study concludes a significant association exists between cigarette smoking with MMP-9 and also relative exposure to circulating inflammation markers plays a potential role in the pathogenesis of CHD.
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ObjectiveTo observe the modulatory effect of modified Zhenwutang on the interleukin-6 （IL-6）， matrix metallopeptidase-9（MMP-9）， type Ⅳ collagen（COL-Ⅳ） in rats with chronic renal failure （CRF） and to investigate the potential mechanism of its treatment of CRF. MethodFifty male SD rats were randomly divided into a modeling group of 40 rats and a normal group of 10 rats， and the modeling group was prepared by continuous adenine gavage for 12 weeks. After successful modelling， the modelling group was divided into the model group， the low dose （7.2 g·kg-1·d-1） group， the medium dose （14.4 g·kg-1·d-1） group， the high dose （28.8 g·kg-1·d-1） group and the Benadryl hydrochloride （10 mg·kg-1·d-1） group for gavage according to the random number table method， In the normal group and the model group， equal volume of distilled water was administered by gavage for 4 weeks. After the administration， the levels of blood creatinine （SCr）， blood urea nitrogen （BUN） and 24 h urine protein （24 h-UTP） were measured， the levels of serum IL-6 were measured by enzyme linked immunosorbent assay（ELISA）. Immunohistochemistry （IHC） was used to detect intercellular cell adhesion molecule-1 （ICAM-1）， IL-6， MMP-9， and other molecules in the rat kidney. The expression of ICAM-1 mRNA， IL-6 mRNA， MMP-9 mRNA and COL-Ⅳ mRNA in rat kidney tissues was measured by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The expression levels of ICAM-1， IL-6， MMP-9 and COL-Ⅳ in rat kidney tissues were measured by Western blot. ResultCompared with the normal group， the levels of SCr， BUN and 24 h-UTP were significantly increased in the model group （P<0.01）； the serum IL-6 level was significantly increased （P<0.01）， the tubular lumen was dilated with atrophy， the tubular epithelial cells were necrotic， swollen and vacuolated， the interstitium was infiltrated by a large number of inflammatory cells and collagen fibers were deposited， the levels of IL-6， ICAM-1 and COL-Ⅳ were strongly positive in the tubular interstitium of the model group （P<0.01）， The levels of ICAM-1 mRNA， IL-6 mRNA and COL-Ⅳ mRNA were significantly increased （P<0.01） and MMP-9 mRNA was significantly decreased （P<0.05） in the model rats. ICAM-1， IL-6 and COL-Ⅳ protein expression was significantly increased （P<0.01） and MMP-9 protein expression was significantly increased （P<0.01） in the renal tissue， and MMP-9 protein expression was significantly decreased （P<0.01）. Compared with the model group， the 24 h-UTP， SCr and BUN levels of rats were significantly reduced after treatment with modified Zhenwutang （P<0.01）， the serum IL-6 level was significantly decreased （P<0.01）， the renal lesions of rats were significantly improved and collagen fiber deposition was reduced； the expression of IL-6， ICAM-1 and COL-Ⅳ in renal tubules and interstitium was weakened， and MMP-9 in ICAM-1 mRNA， IL-6 mRNA and COL-Ⅳ mRNA levels were significantly reduced （P<0.01） and MMP-9 mRNA levels were significantly increased （P<0.05）， ICAM-1， IL-6 and COL-Ⅳ protein expression was significantly reduced （P<0.01） and MMP-9 protein expression was significantly The expression of ICAM-1， IL-6 and COL-Ⅳ proteins was significantly decreased （P<0.01） and MMP-9 protein expression was significantly increased （P<0.01）. ConclusionModified Zhenwutang may regulate the IL-6/MMP-9/COL-Ⅳ signaling pathway， thereby reducing proteinuria， improving renal function， reducing renal pathological damage and delaying the progression of CRF interstitial fibrosis.
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Objective: To investigate the main mechanisms of pulmonary fibrosis following silica nanoparticles (SiNPs) exposure through constructing the macrophage-fibroblast model in vitro, which simulated the process of pulmonary fibrosis. Methods: In January 2021, human mononuclear leukemia cells (THP-1) were treated with 0, 25, 50, 100 μg/ml SiNPs for 24 h. The supernatant of THP-1 cells was collected and applied to human embryonic lung fibroblast cells (MRC-5) which divided into control and low, medium and high dose groups at the logarithmic growth stage for 24 h. MRC-5 cell viability was detected by CCK8. The hydroxyproline (Hyp), interleukin 6 (IL-6), interleukin 1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) expression were detected in the supernatants of MRC-5. The changed proteins were detected by liquid-phase mass spectrometry in high dose group. GeneCard database were applied to identity the differential pulmonary fibrosis proteins in high dose group. Gene Ontology (GO) was performed to identity the key biological process in differential pulmonary fibrosis proteins of high dose group. The String database was used to construct the protein-protein interactions (PPI) network of differential pulmonary fibrosis proteins. The APP of CytoHubba was applied to calculate the key protein of differential pulmonary fibrosis proteins in PPI network. Correlation coefficients between key differential pulmonary fibrosis proteins were calculated using Pearson correlation analysis. Western blotting was applied to detect the expression of key proteins of differential pulmonary fibrosis proteins in different groups. Results: CCK8 results showed that MRC-5 cell viability was increasing in low, medium and high dose groups compared with control group (P<0.05). The expression levels of Hyp and IL-1β in different group were increased compared with control group, the expression levels of IL-6 and TNF-α were increased in high dose group compared with control group (P<0.05). GeneCard database identified 26 differential pulmonary fibrosis proteins, which were mainly involved in extracellular matrix hydrolysis, cell inflammatory response, tissue repair, cell proliferation, inflammation response by GO analysis. The APP of CytoHubba was calculated that matrix metalloproteinase 9 (MMP9) and tissue inhibitor metalloproteinase 1 (TIMP1) played an important role in PPI network. The results of correlation analysis showed that MMP9 was correlated with the expression of matrix metalloproteinase 1 (MMP1), matrix metalloproteinase 3 (MMP3), TIMP1 and epidermal growth factor receptor (EGFR) (r=0.97, 0.98, 0.94, 0.93, P<0.05). Western blotting results showed that TIMP1 protein expression was increased in low, medium and high dose groups, while MMP9 protein expression was increased only in high dose group (P<0.05) . Conclusion: Differential expression proteins related with pulmonary fibrosis in MRC-5 cells mainly regulate biological processes of extracellular matrix hydrolysis, tissue repair, and cellular inflammation response following SiNPs exposure. MMP9 and TIMP1 may be the key proteins, which affected the fibrosis process in vitro pulmonary fibrosis model.
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Objective @#To investigate the effect of isoprene cysteine carboxymethyltransferase (ICMT) gene on the migration and invasion of salivary adenoid cystic cancer cells (SACC) and the related mechanism, to provide experimental evidence for molecular targeted therapy of SACC.@*Methods@# Adenoid cystic cancer cells SACC-LM and SACC-83 were cultured in vitro, and siRNA was transfected into human SACC-LM and SACC-83 cells (experimental group) by transient transfection of a liposome vector. A blank control group and negative control group were set up respectively (transfected NC-siRNA). qRT-PCR was peformed to measure the mRNA expression of ICMT and RhoA in each group after transfection and to determine the silencing efficiency. The expression of ICMT, membrane RhoA, total RhoA, matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and Rho associated with coiled helical binding protein kinase 1 (ROCK1) in each group was detected by Western blot. The proliferation abilityies of SACC cells was detected by CCK-8 assay. The migration and invasion ability of SACC cells were detected by comparing the relative healing area of cell scratch assay and the number of Transwell assay cells. @*Results@#After transfection of ICMT-siRNA into SACC-LM and SACC-83 cells, the expression of ICMT gene and protein in the experimental group was significantly decreased compared with the negative control group and blank control group (P<0.05), but there were no significant differences in the expression of RhoA gene and total protein among all groups (P>0.05). The expression of RhoA membrane proteins, ROCK1, MMP-2, MMP-9 in the experimental group was significantly decreased compared with that in the negative control group and blank control group (P<0.05). Cell proliferation ability was significantly decreased (P<0.05). The migration and invasion abilities were significantly decreased (P<0.05). @*Conclusion @#In vitro silencing of ICMT gene can effectively inhibit the migration and invasion of human SACC-LM and SACC-83 cells, and the mechanism may be related to RhoA-ROCK signaling pathway.
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Objective: To investigate the effect of nuclear factor erythroid 2-related factor 2 (Nrf2) in the alteration of tight junction protein expression in choroid plexus epithelial cells created by lanthanum-activated matrix metalloproteinase 9 (MMP9) . Methods: In October 2020, immortalized rat choroid plexus epithelial cell line (Z310) cells were used as the blood-cerebrospinal fluid barrier in vitro, and were divided into control group and 0.125, 0.25, 0.5 mmol/L lanthanum chloride (LaCl(3)) treatment group. After treating Z310 cells with different concentrations of LaCl(3) for 24 hours, the morphological changes of Z310 cells were observed under inverted microscope, the protein expression levels of MMP9, occludin and zonula occludens-1 (ZO-1) were observed by cellular immunofluorescence method, and the protein expression levels of MMP9, tissue inhibitors of metalloproteinase1 (TIMP1) , occludin, ZO-1 and Nrf2 were detected by Western blotting. The level of reactive oxygen species (ROS) in cells was detected by flow cytometry. Results: Compared with the control group, Z310 cells in the LaCl(3) treatment group were smaller in size, with fewer intercellular junctions, and more dead cells and cell fragments. The expression level of MMP9 protein in cells treated with 0.25 and 0.5 mmol/L LaCl(3) was significantly higher than that in the control group (P<0.05) , and the expression level of TIMP1 and tight junction proteins occudin and ZO-1 was significantly lower than that in the control group (P<0.05) . Compared with the control group, the ROS production level in the 0.25, 0.5 mmol/L LaCl(3) treatment group was significantly increased (P<0.05) , and the Nrf2 protein expression level in the 0.125, 0.25, 0.5 mmol/L LaCl(3) treatment group was significantly decreased (P<0.05) . Conclusion: Lanthanum may increase the level of ROS in cells by down regulating the expression of Nrf2, thus activating MMP9 to reduce the expression level of intercellular tight junction proteins occludin and ZO-1.
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Rats , Animals , Matrix Metalloproteinase 9/metabolism , NF-E2-Related Factor 2/metabolism , Tight Junction Proteins/metabolism , Occludin/pharmacology , Choroid Plexus/metabolism , Reactive Oxygen Species/metabolism , Lanthanum/pharmacology , Epithelial Cells , Zonula Occludens-1 Protein/metabolism , Phosphoproteins/pharmacologyABSTRACT
Objective:To investigate the urineinterleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-33 (IL-33) and matrix metalloproteinase-9 (MMP-9) in patients with IgA nephropathy (IgAN) and their predictive value for disease progression.Methods:110 IgAN patients admitted to Zhuozhou Hospital from January 2018 to October 2021 were selected and divided into IgAN progression group (39 cases) and IgAN non progression group (71 cases) according to the progress of IgAN patients. According to the Oxford Classification Standard System (MEST-C) of IgAN, they were divided into MEST-C≥3 group (42 cases) and MEST-C<3 group (68 cases). According to the estimated glomerular filtration rate (eGFR), they were divided into eGFR≥50 ml/(min·1.73 m 2) group (group A, 63 cases) and eGFR<50 ml/(min·1.73 m 2) group (group B, 47 cases). According to the amount of urinary protein, they were divided into urinary protein≥1.5 g/24 h group (66 cases) and urinary protein<1.5 g/24 h group (44 cases). Multivariate logistic regression was used to analyze the risk factors affecting the progression of IgAN. Receiver operating characteristic (ROC) curve was drawn to analyze the value of urinary IL-6, IL-8, IL-33 and MMP-9 levels in predicting the progression of IgAN. Results:The urinary IL-6, IL-8, IL-33 and MMP-9 levels in IgAN progression group were significantly higher than those in IgAN non progression group, while the serum albumin, eGFR and complement C3 in the IgAN progression group were lower than those in the IgAN non progression group (all P<0.05). The urinary IL-6, IL-8, IL-33 and MMP-9 levels in the MEST-C≥3 group were significantly higher than those in the MEST-C<3 group (all P<0.001). The urinary IL-6, IL-8, IL-33 and MMP-9 levels in the eGFR<50 ml/(min·1.73 m 2) group were significantly higher than those in the eGFR≥50 ml/(min·1.73 m 2) group (all P<0.001). The urinary IL-6, IL-8, IL-33 and MMP-9 in the urinary protein≥1.5 g/24 h group were significantly higher than those in the urinary protein<1.5 g/24 h group (all P<0.001). Multivariate logistic regression analysis showed that the course of disease, serum albumin, eGFR, urinary protein, IL-6, IL-8, IL-33 and MMP-9 were risk factors affecting the progression of IgAN (all P<0.005). The ROC curve showed that the area under the curve (AUC) of IL-6, IL-8, IL-33 and MMP-9 in predicting the progression of IgAN was 0.956 (95% CI: 0.891-0.998), and the sensitivity and specificity were 98.4% and 86.2%, respectively. Conclusions:The elevated levels of urinary IL-6, IL-8, IL-33 and MMP-9 are closely related to the progress of IgAN, and the combination of these four indicators has a good value in predicting the progress of IgAN.
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Objective:To investigate the value of serum thymidine kinase-1 (TK1) and matrix metalloproteinase-9 (MMP-9) level in prognosis evaluation of patients with primary hepatic carcinoma (PHC).Methods:100 PHC patients treated in Panjin Central Hospital from December 2015 to December 2017 were retrospectively selected and divided into survival group ( n=73) and death group ( n=27) according to the prognosis. The clinical characteristics and serum TK1 and MMP-9 levels of the two groups were compared. The relationship between serum TK1 and MMP-9 levels and the clinical characteristics and prognosis of PHC was analyzed. Receiver operating characteristic (ROC) curve was drawn to analyze the value of TK1 and MMP-9 in evaluating the prognosis of PHC. Results:The proportion of multiple lesions, low differentiation, tumor node metastasis (TNM) stage Ⅲ-Ⅳ, extrahepatic metastasis and microvascular invasion in the survival group were lower than those in the death group, and the levels of serum TK1 and MMP-9 were also lower than those in the death group (all P<0.05); The levels of serum TK1 and MMP-9 had no significant correlation with gender, age, tumor length and diameter and child Pugh liver function grade of PHC patients (all P>0.05), but were closely related to the number of lesions, degree of differentiation, TNM stage, extrahepatic metastasis and microvascular invasion (all P<0.05). The areas under the ROC curve of serum TK1 and MMP-9 levels predicting the prognosis of PHC were 0.859 and 0.830. The 3-year survival rate of PHC patients with high level of TK1 and MMP-9 was significantly lower than that of low level patients (all P<0.05). Conclusions:The serum TK1 and MMP-9 levels are correlated with the condition and prognosis of patients with PHC, and can be used as reference indexes for early prognosis evaluation.
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Objective:To investigate the application value of early and late interventional embolization in intracranial aneurysms.Methods:Eighty-two patients with intracranial aneurysm who received treatment in Wenzhou People's Hospital from October 2015 to February 2020 were included in this study. These patients were divided into early (≤ 3 days) and late (> 3 days) groups, with 41 patients in each group, according to time from disease onset to surgery. The early group was subjected to early interventional embolization, and the late group was treated with late interventional embolization. The effects of embolization and National Institutes of Health Stroke Scale score pre- and post-treatment, as well as modified Barthel index, Mini-Mental State Exam score, matrix metalloproteinase-9 level, and soluble intercellular adhesion molecule-1 level post-treatment and prognosis were compared between the two groups.Results:The embolization effects in the early group were statistically superior to those in the late group ( P = 0.046). After treatment, National Institutes of Health Stroke Scale score in the early group was significantly lower than that in the late group [(4.02 ± 1.64) points vs. (6.81 ± 2.02) points, t = 6.86, P < 0.01]. Mini-Mental State Exam score and modified Barthel index in the early group were (28.09 ± 1.35) points and (81.12 ± 9.67) points, respectively, which were significantly higher than (26.01 ± 1.19) points and (73.02 ± 8.19) points in the late group ( t = 7.40, 4.09, both P < 0.001). After treatment, matrix metalloproteinase-9 and soluble intercellular adhesion molecule-1 levels in the early group were (420.33 ± 29.40) μg/L and (403.70 ± 23.28) ug/L, respectively, which were significantly lower than (491.30 ± 31.19) μg/mL and (496.37 ± 30.46) μg/L in the late group ( t = 10.60, 15.47, both P < 0.001). Prognosis in the early group was superior to that in the late group ( P = 0.049). Conclusion:Early interventional embolization has better efficacy than late interventional embolization in the treatment of intracranial aneurysm. The former can effectively improve neurological function and mental state, enhance living ability, and improve prognosis, which may be related to the regulation of matrix metalloproteinase-9 and soluble intercellular adhesion molecule-1 levels.
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Objective:To investigate the correlation and predictive value of serum miR-149-5p and matrix metalloproteinase-9 (MMP-9) and hemorrhagic transformation (HT) after intravenous thrombolysis in patients with acute ischemic stroke (AIS).Methods:Patients with AIS received intravenous thrombolytic therapy in Shangqiu First People's Hospital from September 2019 to February 2022 were enrolled prospectively. They were divided into HT group and non-HT group according to whether HT occurred after intravenous thrombolysis. Serum miR-149-5p and MMP-9 were measured by real-time fluorescence quantitative polymerase chain reaction and enzyme-linked immunosorbent assay respectively. Multivariate logistic regression analysis was used to determine the independent risk factors for HT after thrombolysis. The receiver operating characteristic (ROC) curve was used to evaluate the predictive value of serum miR-149-5p, MMP-9 and their combination for HT after intravenous thrombolysis. Results:A total of 358 patients with AIS received intravenous thrombolytic therapy were enrolled, 71 of them (19.83%) developed HT. The serum MMP-9 in the HT group was significantly higher than that in the non-HT group (273.95±35.23 μg/L vs. 202.71±30.52 μg/L; t=17.062, P<0.001), while the serum miR-149-5p was significantly lower than that in the non-HT group (0.26±0.06 vs. 1.03±0.15; t=42.387, P<0.001). Multivariate logistic regression analysis showed that atrial fibrillation (odds ratio [ OR] 2.282, 95% confidence interval [ CI] 1.731-3.008; P<0.001), time from onset to intravenous thrombolysis ( OR 2.334, 95% CI 1.458-3.735; P<0.001), miR-149-5p ( OR 1.758, 95% CI 1.142-2.705; P=0.010) and MMP-9 ( OR 1.535, 95% CI 1.106-2.129; P=0.010) were the independent risk factors for HT after intravenous thrombolysis. Serum miR-149-5p (area under the curve 0.856, 95% CI 0.803-0.909; when the optimal cut-off value was 0.741, the sensitivity was 80.3% and the specificity was 89.9%), MMP-9 (area under the curve 0.875, 95% CI 0.821-0.929; when the optimal cut-off value was 240.051 μg/L, the sensitivity was 83.1% and the specificity was 90.2%) and their combination (area under the curve 0.897, 95% CI 0.854-0.941; sensitivity 84.5% and specificity 90.6%) had better predictive value for HT after thrombolysis, and there were no significant differences in the predictive value among the three. Conclusions:After intravenous thrombolysis, the serum miR-149-5p is lower and MMP-9 is higher at admission in patients with HT in patients with AIS. Both of them and their combination have better predictive value for HT after intravenous thrombolysis.
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Abstract Background: Interstitial lung disease (ILD) is a common pulmonary complication of connective tissue disease (CTD). This study aims to evaluate the clinical diagnostic value of matrix metalloproteinase-9 (MMP-9), surfactant protein-D (SP-D), and vascular endothelial growth factor (VEGF) as potential biomarkers for CTD-ILD. Methods: This research included 33 CTD-ILD patients, 31 CTD patients without ILD, and 24 healthy control subjects. Then, the value of biomarkers for the diagnosis and evaluation of CTD-ILD was assessed through high-resolution computed tomography (HRCT) findings and pulmonary function test (PFT) parameters. Results: The serum MMP-9, SP-D, and VEGF levels in the CTD-ILD group were higher than those in the CTD-NILD group and healthy group. The ROC curve indicates that VEGF has good to excellent diagnostic performance in diagnosing CTD-ILD, the cut-off that best optimizes sensitivity and specificity in diagnosing CTD-ILD is 277.60 pg/ml (sensitivity, 87.9%; specificity, 83.6%), with an area under the curve (AUC) of 0.905 (95% confidence interval (CI) 0.842-0.968); The ROC curve for MMP-9 suggests this biomarker is fair for diagnosis of CTD-ILD(sensitivity, 81.8%; specificity, 81.8%), with an AUC of 0.867 (95% CI 0.784-0.950), but SP-D only provided lower specificity with higher sensitivity in diagnosing CTD-ILD(sensitivity, 90.9%; specificity, 40.0%). The different serum biomarkers are more specific and sensitive when combined to diagnose ILD. The semiquantitative score for the degree of ILD severity on HRCT was positively correlated with SP-D and VEGF levels ( r = 0.461, P = 0.007; r = 0.362, P = 0.039), and serum MMP-9 levels were elevated in the UIP subgroup compared to the non-UIP subgroup. The percentage of diffusing capacity of the lung for carbon monoxide (DLco) (% predicted) had a negative correlation with the SP-D level ( r = − 0.407, P = 0.044) and a statistically negative correlation between MMP-9 and the forced vital capacity (FVC) ( r = − 0.451, P = 0.024). Conclusions: Serum MMP-9, SP-D, and VEGF levels may have clinical value in screening and evaluating the severity of CTD-ILD. Key points Serum MMP-9, SP-D, and VEGF levels were increased in patients with CTD-ILD and they may have clinical value in screening and evaluating the severity of CTD-ILD. Serum SP-D and VEGF levels had a positive correlation with ILD severity as measured using semiquantitative HRCT scores. Serum MMP-9 levels were elevated in the UIP subgroup compared to the non-UIP subgroup. Therefore, further research is required to determine the role of serum MMP-9 levels in the preliminary determination of the ILD subtype. Serum MMP-9 levels had a negative correlation with DLco, and serum SP-D levels had a negative correlation with FVC.
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ABSTRACT Purpose Quantify the tissue content of metalloproteinase-9 (MMP-9) and collagen in colic mucosa with and without intestinal transit after infliximab administration in rats subjected to Hartmann's surgery. Methods Twenty-two rats underwent colon diversion by Hartmann's surgery. Animals were maintained with intestinal bypass for 12 weeks to induce development of diversion colitis (DC). Afterwards, animals were divided into three groups: first group received subcutaneous application of saline solution (SS) 0.9%, while the remaining two groups received infliximab subcutaneously at doses of 5 or 10 mg·kg-1·week-1 for five consecutive weeks. After the intervention, animals were sacrificed, removing the segments with and without intestinal transit. Diversion colitis was diagnosed by histological study, and its intensity was determined by a validated inflammatory scale. Tissue expression of MMP-9 was assessed byimmunohistochemistry, while total collagen was assessed by histochemistry. Tissue content of both was measuredby computerized morphometry. Results Colon segments without intestinal transit had a higher degree of inflammation, which improved in animals treated with infliximab. Collagen content was always lower in those without intestinal transit. There was an increase in the collagen content in the colon without transit in animals treated with infliximab, primarily at a dose of 10 mg·kg-1·week-1. There was an increase in the content of MMP-9 in the colon without fecal transit, and a reduction was observed in animals treated with infliximab, regardless of the dose used. Conclusions Application of infliximab reduces inflammation, increases the total collagen content and decreases the content of MMP-9 in the colon without intestinal transit.
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Animals , Rats , Colon/surgery , Intestinal Mucosa , Collagen , Rats, Wistar , Metalloproteases , InfliximabABSTRACT
Objective:To investigate the changes of the plasma matrix metalloproteinase (MMP)-2 and MMP-9 levels and their clinical significances during the course of concurrent chemoradiotherapy (CCRT) in nasopharyngeal carcinoma (NPC) patients.Methods:From January 2018 to June 2019, 46 patients with nasopharyngeal carcinoma were treated in the department of oncology, Changsha Central Hospital Affiliated to Nanhua University. All patients were confirmed by pathology. They were divided into early NPC group ( n=32) and invasive NPC group ( n=16) according to the degree of invasion. The early NPC group was treated with concurrent chemoradiotherapy alone, and the invasive NPC group was treated with neoadjuvant chemotherapy combined with concurrent chemoradiotherapy. Blood samples were collected at four stages of the treatment, and the concentrations of MMP-2 and MMP-9 were detected by enzyme linked immunosorbent assay (ELISA). Results:The longer the treatment time, the lower the concentration of MMP-9 ( P=0.007) in early NPC group; There was no significant difference in MMP-9 level before treatment, after neoadjuvant chemotherapy, after concurrent chemoradiotherapy, at the end of treatment and the first follow-up ( P>0.05) in invasive NPC group. There was no significant difference in the content of MMP-2 between the two groups before and after treatment ( P>0.05). There was no correlation between serum MMP-2 and MMP-9 levels and tumor stage, lymph node metastasis, tumor invasion and response rate ( P>0.05) in invasive NPC patients, while the level of MMP-9 was positively correlated with white blood count (WBC) and neutrophil count ( r=0.85, P=0.004, r=0.82, P=0.003); The ratio of MMP-9/MMP-2 was positively correlated with WBC and neutrophil count ( r=0.86, P=0.003, r=0.83, P=0.001). Conclusions:Synchronous radiotherapy and chemotherapy can reduce the serum MMP-9 level in early stage NPC patients, but it has no effect on the serum MMP-9 level in patients with invasive NPC, which suggests that synchronous radiotherapy and chemotherapy can not prevent the proliferation and distant metastasis of cancer cells in patients with invasive NPC.
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Objective:To investigate the changes and clinical significance of serum matrix metalloproteinase-9 (MMP-9), squamous cell carcinoma antigen (SCC), cytokeratin 19 fragment (CYFRA21-1), carcinoembryonic antigen (CEA) and neuron-specific enolase (NSE) in peripheral lung cancer.Methods:Sixty-eight patients with peripheral lung cancer who received treatment in Luqiao Hospital of Taizhou Enze Medical Center (Group) between January 2017 and January 2020 were included in the observation group. Sixty-five patients with benign lung diseases who concurrently received treatment in the same hospital were included in the observation group 1, and another 65 healthy participants who concurrently received physical examination were included in the control group. Serum levels of MMP-9, CYFRA21-1, SCC, NSE and CEA were compared among the three groups. The sensitivity and specificity of using these indicators alone and in combination in the diagnosis of peripheral lung cancer were compared.Results:Serum levels of MMP-9, CYFRA21-1, SCC, NSE and CEA in the observation group (14.98 ± 2.10) ng/mL, (17.13 ± 2.71) ng/mL, (1.98 ± 0.41) μg/mL, (24.13 ± 2.10) ng/mL and (17.10 ± 2.10) ng/mL, respectively, which were significantly higher than those in the observation group 1 [(9.12 ± 1.41) ng/mL, (10.12 ± 1.58) ng/mL, (1.37 ± 0.31) μg/mL, (16.31 ± 1.78) ng/mL, (12.13 ± 1.79) ng/mL] and control group [(5.10 1 ± 0.68) ng/mL, (6.02 ± 0.94) ng/mL, (0.71 ± 0.11) μg/mL, (11.10 ± 1.02) ng/mL, (8.13 ± 1.02) ng/mL] ( F1 = 932.781, F2 = 737.100, F3 = 368.591, F4 = 989.851, F5 = 462.291, all P < 0.05). Serum levels of MMP-9, CYFRA21-1, SCC, NSE and CEA in patients with stage I-II peripheral lung cancer were (11.12 ± 2.10) ng/mL, (9.12 ± 1.85) ng/mL, (1.52 ± 0.21) μg/mL, (18.12 ± 3.02) ng/mL, (7.52 ± 1.02) ng/mL, respectively, which were significantly lower than those in patients with stage III-IV peripheral lung cancer [(15. 89 ± 2.18) ng/mL, (21.56 ± 2.11) ng/mL, (2.04 ± 0.31) μg/mL, (28.15 ± 2.62) ng/mL, (15.12 ± 1.55) ng/mL, t1 = 9.013, t2 = 25.146, t3 = 7.714, t4 = 14.586, t5 = 22.705, all P < 0.05]. The sensitivity (83.33%) and specificity (86.67%) of combined detection of all indicators were significantly higher than those of single detection of MMP-9 (50.00%, 59.68%), CEA (50.00%, 61.29%), CYFRA21-1 (66.67%, 58.06%), SCC (50.00%, 54.84%) or NSE (66.67%, 58.06%) (all P < 0.05). Conclusion:Serum levels of MMP-9, CYFRA21-1, SCC, NSE and CEA in patients with peripheral lung cancer are significantly increased, which has an important value in the diagnosis of peripheral lung cancer. The combined detection of the above indicators can increase the diagnostic accuracy of peripheral lung cancer in the clinic.
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Objective:To investigate the therapeutic effect and mechanism of modified Wenjingtang on endometriosis （EM） rats with kidney deficiency and blood stasis. Method:Healthy non-pregnant female Sprague-Dawley （SD） rats of SPF grade were randomly divided into the blank group and experimental group. After being modeled via soaking in ice water and subcutaneous injection of epinephrine hydrochloride， the ones in the experimental group were further divided into the sham operation group and EM model group， with the former only undergoing laparotomy and the latter further receiving autologous endometrial transplant for triggering EM. The successfully modeled rats with EM due to kidney deficiency and blood stasis were randomized into the positive drug （danazol， 63 mg·kg<sup>-1</sup>） group and low- （5 g·kg<sup>-1</sup>）， medium- （10 g·kg<sup>-1</sup>）， and high-dose （20 g·kg<sup>-1</sup>） modified Wenjingtang groups. The corresponding drugs were administered by gavage， once per day， for four weeks. Then the ectopic and eutopic endometrial tissues were stained with hematoxylin-eosin （HE） to observe the histopathological changes. The protein and mRNA expression levels of cysteinyl aspartate-specific proteinase-8 （Caspase-8）， matrix metalloproteinase-9 （MMP-9）， N-cadherin， and E-cadherin were detected by immunohistochemistry （IHC）， Western blotting， and real-time polymerase chain reaction （Real-time PCR）， respectively. Result:The IHC and Western blot revealed that the protein expression levels of MMP-9 and N-cadherin in eutopic and ectopic endometrial tissues of the model group were significantly increased as compared with those in the sham operation group （<italic>P</italic><0.01）， while the levels of Caspase-8 and E-cadherin was significantly decreased （<italic>P</italic><0.01）. Compared with the model group， the danazol and low-， medium-， and high-dose modified Wenjingtang groups exhibited obviously down-regulated MMP-9 and N-cadherin protein expression in eutopic and ectopic endometrial tissues （<italic>P</italic><0.05，<italic>P</italic><0.01）， but up-regulated Caspase-8 and E-cadherin （<italic>P</italic><0.05， <italic>P</italic><0.01）. Real-time PCR uncovered that the mRNA expression levels of MMP-9 and N-cadherin in eutopic and ectopic endometrial tissues of the model group were significantly elevated as compared with those in the sham operation group （<italic>P</italic><0.01）， whereas the levels of Caspase-8 and E-cadherin significantly declined （<italic>P</italic><0.01）. The comparison with the eutopic endometrial tissue in the model group showed that the mRNA expression levels of MMP-9 and N-cadherin in the danazol group and high- and medium-dose modified Wenjingtang groups were significantly down-regulated， while those of Caspase-8 and E-cadherin were significantly up-regulated （<italic>P</italic><0.05，<italic>P</italic><0.01）. Conclusion:Modified Wenjingtang alleviates the immunosuppression and blocks the angiogenesis in EM rats with kidney deficiency and blood stasis syndrome by regulating the expression of such cytokines as Caspase-8， MMP-9， N-cadherin， and E-cadherin， thus exerting the therapeutic effect against EM. The above-mentioned micro-indicators are potential markers reflecting the disease （EM）， syndrome （kidney deficiency and blood stasis）， and pathological mechanisms （immunosuppression and angiogenesis）.
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@#AIM: To investigate the effects of doxycycline(DOX)on vasculogenic mimicry(VM)in human pterygium fibroblasts(HPFs)and its molecular mechanisms.<p>METHODS: Primary cultured HPFs were identified by Vimentin and CK through immunocytochemical staining. HPFs were divided into control group and DOX group including low, medium and high concentrations(50, 100, 200mg/L). The activity and migration of HPFs were detected by cell counting kit-8(CCK-8)and wound healing assay. The density of VM was observed by three-dimensional cell culture and periodic acid schiff(PAS)staining and compared the differences of VM formation in each group. Western blot was used to analyze the expression of matrix metalloproteinase-9(MMP-9)and vascular endothelial growth factor(VEGF).<p>RESULTS:Immunocytochemical staining results showed that the cells were spindle shaped, meanwhile, they were positive for Vimentin and negative for CK, which were consistent with the characteristics of fibroblasts. Compared with the control group, the cell activity, mobility, VM density and the expression of MMP-9 and VEGF proteins in the DOX group were significantly decreased(<i>P</i><0.05). Compared among different concentrations of DOX groups, the differences were statistically significant(<i>P</i><0.05). Correlation analysis indicated that VM density formed by HPFs was significantly positively correlated with the protein expression of MMP-9 and VEGF(<i>r</i>=0.949, 0.960, all <i>P</i><0.05).<p>CONCLUSION: DOX can inhabit HPFs activity, migration, VM density by reducing the expression of MMP-9 and VEGF, suggesting that MMP-9 and VEGF may be the molecular mechanisms of VM formation in pterygium.
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Objective:To investigate the expression and correlation of H19, matrix metalloproteinase (MMP)-2 and MMP-9 in patients with recurrent spontaneous abortion (RSA).Methods:Human extravillous trophoblast cell line HTR-8 was cultured in vitro. Lentivirus was used to infect the HTR-8 cell line to over-express or knockdown the expression of H19. The concentrations of MMP-2 and MMP-9 protein in cell culture supernatant were detected by enzyme linked immunosorbent assay (ELISA). The expression levels of H19, MMP-2 and MMP-9 mRNA in villi of patients with RSA were detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Spearman correlation analysis was used to understand the correlation between H19 and the expression levels of MMP-2 and MMP-9. Results:After overexpression of H19, the expression levels of MMP-2 and MMP-9 mRNA and protein concentration in Lv-ph19 group were significantly higher than those in Lv-vector group ( P<0.05); After interfering with the expression of H19, the expression levels of MMP-2 and MMP-9 mRNA and protein concentration in Lv-shH19 group were significantly lower than those in Lv-shcon control group ( P<0.05). The number of spontaneous abortions in patients with recurrent spontaneous abortion was significantly higher than that in the control group ( P<0.05). qRT-PCR showed that the expression levels of H19, MMP-2 and MMP-9 mRNA in villi of patients with RSA were significantly lower than those in the control group ( P<0.05). There was a positive correlation between H19 and the expression levels of MMP-2 and MMP-9 ( P<0.05). Conclusions:H19 regulates the expression of MMP-2 and MMP-9 of trophoblast during early pregnancy, and the abnormal expression of H19, MMP-2, and MMP-9 in human first-trimester villous tissues was related with the incidence of early miscarriage.
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Objective:To investigate the role and mechanism of AT-rich interactive domain 1A gene (ARID1A) in glioblastoma invasion.Methods:Human glioblastoma cell line U87 was cultured in vitro. U87 cells transfected with recombinant pcDNA3.1 (+ )-ARID1A by lipofectamine liposome method as ARID1A overexpression group, and U87 cells transfected with empty plasmid as vector group. Untreated U87 cells were as blank control group. The transfection efficiency was verified by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) and Western blot. Transwell invasion test was used to detect the invasion ability of cells, and Western blot was used to detect the protein expression of c-myc, matrix metalloproteinases (MMP)-2 and MMP-9. Results:48 hours after transfection with ARID1A eukaryotic expression plasmid, the cell invasion ability of ARID1A overexpression group, vector group and blank control group were (42.2±11.5)%, (98.6±4.8)%, (100.0±5.1)%. There was significant difference between ARID1A overexpression group and the other two groups ( P<0.01); The expressions of c-myc, MMP-2 and MMP-9 in ARID1A overexpression group were lower than those in vector group and blank control group ( P<0.01). Conclusions:ARID1A can inhibit the invasion of glioblastoma by inhibiting the expression of MMP-2/MMP-9, and can be used as a potential therapeutic target for glioblastoma.
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RESUMO Objetivo: Determinar se os níveis plasmáticos das metaloproteinases de matriz -2 e -9 tem associação com a mortalidade na unidade de terapia intensiva em pacientes com trauma craniencefálico grave, independentemente de lesões não cerebrais associadas. Métodos: Esta coorte prospectiva incluiu 39 pacientes do sexo masculino com trauma craniencefálico grave (escore na escala de coma Glasgow na admissão hospitalar: 3 - 8). Os níveis plasmáticos das metaloproteinases -2 e -9 foram determinados por ELISA no momento da admissão na unidade de terapia intensiva. Resultados: O trauma craniencefálico grave apresentou mortalidade de 46% na unidade de terapia intensiva. Concentrações mais elevadas de metaloproteinase -9 apresentaram associação com a mortalidade: 147,94 ± 18,00ng/mL para pacientes que sobreviveram e 224,23 ± 23,86ng/mL para os que não sobreviveram (média ± erro padrão, respectivamente; p = 0,022). Todavia, não houve associação significativa entre os níveis de metaloproteinase -2 e a mortalidade na unidade de terapia intensiva: 315,68 ± 22,90ng/mL para o grupo de sobreviventes e 336,55 ± 24,29ng/mL entre os pacientes que não sobreviveram (p = 0,499). Além disso, não se observaram associações significativas entre os níveis de metaloproteinase -2 (p = 0,711) ou metaloproteinase -9 (p = 0,092) e a presença de lesões não cerebrais associadas. Conclusão: Em vítimas de traumatismo craniencefálico grave, níveis elevados de metaloproteinase -9 tiveram valor preditivo para o desfecho fatal na unidade de terapia intensiva independentemente da presença de lesões não cerebrais associadas. Por outro lado, no mesmo cenário, os níveis plasmáticos de metaloproteinase -2 não apresentaram associação com a mortalidade na unidade de terapia intensiva
Abstract Objective: To determine whether the matrix metalloproteinases-2 and -9 plasma levels were associated with intensive care unit mortality in patients who suffered severe traumatic brain injury, despite the presence of extracerebral injuries. Methods: This prospective cohort enrolled 39 male patients who suffered severe traumatic brain injury (Glasgow coma scale: 3 - 8 at hospital admission). The plasma matrix metalloproteinase -2 and matix metalloproteinase -9 levels were determined by ELISA at the time of intensive care unit admission. Results: Severe traumatic brain injury was associated with a 46% intensive care unit mortality rate. Higher plasma matrix metalloproteinase -9 concentrations were associated with mortality: 147.94 ± 18.00ng/mL for survivors and 224.23 ± 23.86ng/mL for nonsurvivors (mean ± standard error of the mean, p = 0.022). In contrast, there was no significant association between matrix metalloproteinase -2 levels and intensive care unit mortality: 315.68 ± 22.90ng/mL for survivors and 336.55 ± 24.29ng/mL for nonsurvivors (p = 0.499). Additionally, there were no significant associations between matrix metalloproteinase -2 (p = 0.711) and matrix metalloproteinase -9 (p = 0.092) levels and the presence of associated lesions. Conclusion: Increased plasma matrix metalloproteinase -9 levels were associated with intensive care unit mortality following severe traumatic brain injury, regardless of the presence of extracerebral injuries. Conversely, in this same context, plasma matrix metalloproteinase -2 levels were not associated with short-term fatal outcome prediction.