Correlation between extracellular MMP-9 polymorphism and genetic susceptibility of PACG in a Han Chinese population / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To analyze the association of extracellular matrix metalloprotease 9(MMP-9)single nucleotide polymorphism(SNP)with genetic susceptibility of primary angle-closure glaucoma(PACG)in a Han Chinese population. <p>METHODS: Totally 200 PACG patients(PACG group)and 200 healthy people(normal control group)were collected in our hospital from January 2014 to December 2016. Peripheral venous blood was collected and extracted for genomic DNA, the polymerase chain reaction-restriction fragment length polymorphism technique(PCR-RFLP)was applied to detect the alleles and genotypes of rs2250889, rs2274755 and rs2664538 sites in MMP-9 gene. The frequency distribution of alleles and genotypes in the two groups were calculated by chi-square test, and its association with genetic susceptibility of PACG was analyzed. <p>RESULTS: There were no statistical differences in age, gender, body mass index, diastolic blood pressure and systolic blood pressure in the two groups(<i>P</i>>0.05). The genotype frequencies of rs2250889, rs2274755 and rs2664538 sites in MMP-9 gene were in line with Hardy-Weinberg equilibrium. The genotype and allele frequency distribution of rs2250889 and rs2664538 sites were significantly different between the PACG group and the normal control group(<i>P</i><0.05), while the genotype and allele frequency distribution of rs2274755 sites in the two groups had no statistical difference(<i>P</i>>0.05). The subjects whose rs2250889 site carrying the CC genotype was susceptible to PACG. Similarly, the rs2664538 site carrying the GG genotype was susceptible to PACG. <p>CONCLUSION: The rs2250889 and rs2664538 polymorphisms of MMM-9 are correlated with the risk of PACG in a Han Chinese population, while the rs2274755 polymorphism is not related to genetic susceptibility of PACG.

Protective effect of nerve growth factor combined with mecobalamine on optic nerve and influence on MMP-2 and TIMP-2 / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the protective effect of nerve growth factor(NGF)combined with mecobalamine on optic nerve and the influence on matrix metalloproteinase-2(MMP-2), tissue inhibitor of matrix metalloprotease-2(TIMP-2). <p>METHODS: Totally 54 patients(73 eyes)with acute angle closure glaucoma undergoing trabeculectomy was included in the study, and the patients were divided into the control group(30 cases, 35 eyes)and combination group(24 cases, 38 eyes)according to the digital table method. The patients in the control group were treated with mecobalamine, and the patients in combination group were treated with NGF combined with mecobalamine. Visual acuity, intraocular pressure, mean light sensitivity(MS), visual field mean defect(MD), retinal nerve fibre layer(RNFL)thickness, the nipple cup/disc ratio, P100 wave incubation period and amplitude of P100 wave, neuron-specific enolase(NSE), nitric oxide(NO), nitric oxide synthase(NOS), matrix metalloproteinase-2(MMP-2), metalloproteinases tissue inhibiting factor-2(TIMP-2)before and after treatment were observed in the two groups. The adverse reactions during medication were supervised. <p>RESULTS: Compared with those before treatment, visual acuity, MS, and P100 wave amplitudes increased, MD and P100 wave latency decreased(<i>P</i><0.05)after treatment, the change ranges in combination group were greater than those in control group(<i>P</i><0.05). There was no significant change in intraocular pressure, RNFL, the nipple cup/disc ratio before and after treatment in the two groups(<i>P</i>>0.05). Compared with those before treatment, serum NO, NOS, MMP-2 levels in the two groups increased, and the levels of NES, TIMP-2, TIMP-2/MMP-2 decreased(<i>P</i><0.05), but the change degrees in combination group were greater than those in control group(<i>P</i><0.05). <p>CONCLUSION: NGF combined with mecobalamine can improve postoperative visual function of glaucoma, which has certain regulatory effects on MMP-2 and TIMP-2.

Serum matrix metalloproteinase-1 level represents disease activity as opposed to fibrosis in patients with histologically proven nonalcoholic steatohepatitis

BACKGROUND/AIMS: Nonalcoholic steatohepatitis (NASH) is prevalent in both economically developed and developing countries. Twenty percent of NASH progresses to cirrhosis with/without hepatocellular carcinoma, and there is an urgent need to find biomarkers for early diagnosis and monitoring progression of the disease. Using immunohistochemical and immunoelectron microscopic examination we previously reported that expression of matrix metalloproteinase-1 (MMP-1) increased in monocytes, Kupffer cells and hepatic stellate cells in early stage NASH. The present study investigated whether serum MMP-1 levels reflect disease activity and pharmaceutical effects in NASH patients. METHODS: We measured the serum levels of MMPs, tissue inhibitors of metalloproteinases (TIMPs), and several cytokines/chemokines in patients with histologically proven early and advanced stages of NASH and compared them with those in healthy controls. RESULTS: Serum MMP-1 levels in stage 1 fibrosis, but not in the more advanced fibrosis stages, were significantly higher than in healthy controls (P=0.019). There was no correlation between serum MMP-1 level and fibrosis stage. Serum MMP- 1 levels in NASH patients represented disease activity estimated by serum aminotransferase values during the follow-up period. In contrast, MMP-2, MMP-9 and TIMPs did not change with disease activity. Consistent with the finding that MMP-1 is expressed predominantly in monocytes and Kupffer cells, serum levels of monocyte chemotactic protein-1 and granulocyte-colony stimulating factor were significantly increased in NASH with stage 1 fibrosis. CONCLUSIONS: These results suggest that serum MMP-1 levels represent disease activity and may serve as a potential biomarker for monitoring the progression of NASH.

Evaluation of efficacy of Zhitong-Jiangu recipes in treatment of cartilage injury in osteoarthritis using MR T2 mapping: An experimental study / 中国介入影像与治疗学

Objective To explore the value of MR T2 mapping imaging in evaluation of the effection of Zhitong-Jiangu recipes in treatment of cartilage injury in osteoarthritis.Methods The models of osteoarthritis in the knee of New Zealand white rabbits were made with intra-articular injection of papain.Totally 48 New Zealand white rabbits were randomly divided into normal control group,model control group,glucosamine hydrochloride treatment group and Zhitong-Jiangu recipes treatment group.MR T2 mapping was performed on the knee and the content of MMP-1 in articular cartilage was analyzed with immunohistochemistry 4 weeks after treatment.The differences of T2 value and MMP-1 content of articular cartilage among the four groups were compared.The correlation among the different intervention methods,MMP-1 content of articular cartilage and the T2 value of articular cartilage were analyzed.Results T2 value of articular cartilage in the model control group was significantly higher than that in the other three groups (all P＜0.01),and there was no significant difference of T2 value among the other three groups.The MMP-1 content of articular cartilage in glucosamine hydrochloride treatment group and Zhitong-Jiangu recipes treatment group was lower than that in model control group (both P＜0.05),and was higher than that in normal control group (both P＜0.05).There was no significant difference between the two groups.The relationship between MMP-1 content and T2 value of articular cartilage was nonlinear.With the increase of MMP-1 content,T2 value first increased slowly and then increased rapidly (P＜0.05).Conclusion MR T2 mapping imaging is helpful to evaluating the efficacy of Zhitong-Jiangu recipes in treatment of cartilage injury in osteoarthritis in rabbit models.The relationship between MMP-1 content and T2 value of articular cartilage may be nonlinear.

Lentivirus-mediated shRNA interference targeting ADAM17 down-regulates MMP-9 expression in airway epithelial cells via TNF-α/NF-κB signaling / 中国药理学通报

Aim To study the role of TNF-α/NF-κB signaling in matrix metalloproteinase ( MMP)-9 expres-sion induced by lipopolysaccharide ( LPS ) in airway epithelial cells, and to investigate the effects of lenti-virus mediated RNAi targeting a disintegrin and metal-loproteinase 17 ( ADAM17 ) gene on MMP-9 expression induced by LPS. Methods The ADAM17 siRNA ex-pression vector was constructed, and packaged to re-combinant lentivirus in 293T cells. The HBE4-E6/E7 cells were pretreated for 30 min by NF-κB inhibitor ( PDTC) and a recombinant human TNFR p75-Fc fu-sion protein ( Etanercept) , or infected by the recombi-nant lentivirus for 72 h, and then stimulated for 24 h by LPS or TNF-α. The release of TNF-α was detected by ELISA. The mRNA and protein levels of MMP-9 were analyzed respectively by RT-PCR and Western blot. NF-κB activity was detected by electrophoretic mobility shift assay. Results LPS and TNF-α signifi-cantly increased MMP-9 mRNA and protein expressions and the activation of NF-κB in HBE4-E6/E7 cells ( P0. 05 ) . And PDTC significantly inhibited MMP-9 mRNA and protein expressions and the activation of NF-κB induced by TNF-α ( P <0. 05 ) . Conclusions TNF-α/NF-κB signaling partic-ipates in the regulation of MMP-9 expression induced by LPS in airway epithelial cells, and lentivirus-media-ted RNAi targeting ADAM17 plays an important role in that signaling pathway upstream by regulating TNF-αrelease.

Chitin from the Extract of Cuttlebone Induces Acute Inflammation and Enhances MMP1 Expression

We previously reported that the extract from cuttlebone (CB) has wound healing effect in burned lesion of rat. In present study, the main component of CB extract was analyzed and its wound healing activity was evaluated by using in vitro acute inflammation model. The extract of CB stimulated macrophages to increase the production of TNF-alpha. The extract also enhanced the production of TGF-beta and VEGF, which were involved in angiogenesis and fibroblast activation. The treatment with CB extract enhanced proliferation of murine fibroblast. CB extract also induced the activation of fibroblast to increase the secretion of matrix metalloproteases 1 (MMP1). The constituent of CB extract which has wound healing activity was identified as chitin by HPLC analysis. The mechanism that the CB extract helps to promote healing of burned lesion is associated with that chitin in CB extracts stimulated wound skins to induce acute inflammation and to promoted cell proliferation and MMP expression in fibroblast. Our results suggest that CB or chitin can be a new candidate material for the treatment of skin wound such as ulcer and burn.

MMP-1/PAR-1 signal transduction axis and its prognostic impact in esophageal squamous cell carcinoma

The matrix metalloprotease-1 (MMP-1)/protease-activated receptor-1 (PAR-1) signal transduction axis plays an important role in tumorigenesis. To explore the expression and prognostic value of MMP-1 and PAR-1 in esophageal squamous cell carcinoma (ESCC), we evaluated the expression of two proteins in resected specimens from 85 patients with ESCC by immunohistochemistry. Sixty-two (72.9 percent) and 58 (68.2 percent) tumors were MMP-1- and PAR-1-positive, respectively, while no significant staining was observed in normal esophageal squamous epithelium. MMP-1 and PAR-1 overexpression was significantly associated with tumor node metastasis (TNM) stage and regional lymph node involvement. Patients with MMP-1- and PAR-1-positive tumors, respectively, had poorer disease-free survival (DFS) than those with negative ESCC (P = 0.002 and 0.003, respectively). Univariate analysis showed a significant relationship between TNM stage [hazard ratio (HR) = 2.836, 95 percent confidence interval (CI) = 1.866-4.308], regional lymph node involvement (HR = 2.955, 95 percentCI = 1.713-5.068), MMP-1 expression (HR = 2.669, 95 percentCI = 1.229-6.127), and PAR-1 expression (HR = 1.762, 95 percentCI = 1.156-2.883) and DFS. Multivariate analysis including the above four parameters identified TNM stage (HR = 2.035, 95 percentCI = 1.167-3.681), MMP-1 expression (HR = 2.109, 95 percentCI = 1.293-3.279), and PAR-1 expression (HR = 1.967, 95 percentCI = 1.256-2.881) as independent and significant prognostic factors for DFS. Our data suggest for the first time that MMP-1 and PAR-1 were both overexpressed in ESCC and are novel predictors of poor patient prognosis after curative resection. The MMP-1/PAR-1 signal transduction axis might be a new therapeutic target for future therapies tailored against ESCC.

Inorganic sulfur reduces the motility and invasion of MDA-MB-231 human breast cancer cells

This study investigated the effects of inorganic sulfur on metastasis in MDA-MB-231 human breast cancer cells. MDA-MB-231 cells were cultured in the absence or presence of various concentrations (12.5, 25, or 50 micromol/L) of inorganic sulfur. Cell motility, invasion, and the activity and mRNA expression of matrix metalloproteases (MMPs) were examined. Numbers of viable MDA-MB-231 cells did not differ by inorganic sulfur treatment from 0 to 50 micromol/L within 48 h. Inorganic sulfur significantly decreased cell motility and invasion in the MDA-MB-231 cells in a dose-dependent manner (P < 0.05), as determined using a Boyden chamber assay and a Matrigel chamber. The activities of MMP-2 and MMP-9 were significantly reduced by inorganic sulfur in a dose-dependent manner (P < 0.05). The inorganic sulfur also significantly inhibited MMP-2 and MMP-9 expression in the cells (P < 0.05). These data suggest that inorganic sulfur can suppress cancer cell motility and invasion by inhibiting MMP-2 and MMP-9 activity and gene expression in MDA-MB-231 cells.

Gene expression of matrix metalloprotease and their inhibitors in liver tissue of rats with Vibrio vulnificus sepsis / 中华临床感染病杂志

Objective To investigate gene expressions of matrix metalloprotease (MMP-2, MMP-9) and tissue inhibitor of metalloproteinase-2 (TIMP-2) and RECK ( reversion inducing cysteine-rich protein with Kazal motifs) in the liver tissue of rats with Vibrio vulnificus sepsis. Methods Vibrio vulnificus sepsis model was induced by injection of 1 mL/100 g Vibrio vulnificus (9 × 106 cfu/mL) at hind limbs in 50 Sprague-Dawley male rats of clean grade, each time 10 animals were sacrificed at 2, 6, 9, 12 and 16 h after injection and liver samples were taken. Ten rats served as control group. The ethological changes were observed. The total RNA was extracted from liver tissue and the gene expressions of MMP-2, MMP-9, TIMP-2 and RECK were evaluated by semi-quantitative RT-PCR. Results The rats manifested shortness of breath, fever and swelling in hind limbs at 4 h after injection of Vibrio vulnificus. These symptoms gradually deteriorated, and the rats presented cyanosis and convulsion at 12 h. The gene expressions of MMP-2 and MMP-9 were markedly up-regulated, while those of TIMP-2 and RECK were down-regulated. Conclusion The study suggests that the elevation of MMP-2 and MMP-9 and down-regulation of TIMP-2 and RECK is associated with the development of Vibrio vulnificus sepsis.

Effect of High Dose External Irradiation on the Matrix Metalloprotease-2 Expression in a Rat Carotid Artery Injury Model

BACKGROUND AND OBJECTIVES: Remodeling of the injured arterial wall is dependent on the action of several extracellular proteases, including matrix metalloprotease-2 (MMP-2), and this protein is associated with the migration of vascular smooth muscle cells. The effect of a high dose of external irradiation (20 Gy) on the MMP-2 expression in neointimal hyperplasia is not known. MATERIALS AND METHODS: Sprague-Dawley rats were subjected to balloon injury to the common carotid artery. At 24 hours after injury, 20 Gy external irradiation was done for the irradiated group (n=25) and this was not done for the control group (n=25). The percent area stenosis, the maximal intimal thickness, the intima/media area ratio on H-E staining and the MMP-2 positivity on the immunohistochemical staining were measured. Western blotting and a gelatin zymogram for determining the MMP-2 protein expression were also performed after the injury. RESULTS: The parameters of neointimal hyperplasia such as the percent area stenosis, the maximal intimal thickness and the intima/media area ratio were 40.2+/-12.1%, 0.30+/-0.12 mm and 1.27+/-0.32, respectively, at 14 days after injury, and these parameters were maintained as a hyperplastic state at 28 days after injury in the control group. There was undetectable neointimal hyperplasia in the irradiated group compared with the control group (p<0.01). Western blotting demonstrated an increase in the MMP-2 protein level beginning 2 to 4 days after injury in the control group, but there was only a transient increase in the MMP-2 level at day 2 after injury in the irradiated group. The gelatin zymogram and immunohistochemical staining also showed the expression of MMP-2 in the control group, but not in the irradiated group. CONCLUSION: These findings suggest the suppressed expression of MMP-2 is associated with reduced neointimal hyperplasia in the balloon injury-rat model.