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AIM: To investigate the expression changes of MMP-12 during the long-term axon regeneration induced by the lens injury after the optic nerve clamp trauma in sprague-dawley(SD)rats.METHODS: The optic nerve injury model and lens injury model of SD rats were established, and the 24 experimental animals were divided into control group; lens injury group; optic nerve injury group; lens injury combined with optic nerve injury group, with 6 rats in each group. Reference transcriptome sequencing was used to analyze the expression changes of differentially expressed genes in the injured optic nerve region, and relevant differentially expressed genes with high expression were screened. Quantitative real-time polymerase chain reaction(qRT-PCR)and enzyme-linked immunosorbent assay(ELISA)were used to quantify the expression changes of matrix metalloproteinase-12(MMP-12)in the injured optic nerve region.RESULTS: The Principal Component Analysis of transcriptome sequencing indicated that lens injury combined with optic nerve injury was the principal component of gene expression change. Analysis of gene expression differences showed that the expression of MMP-12 gene was up-regulated in the lens injury combined with optic nerve injury group. The mRNA expression level of MMP-12 in the lens injury combined optic nerve injury group was up-regulated compared with the control group, the optic nerve injury group and the lens injury group at 14d and 21d after successful modeling(P<0.05). At 7, 28d, there was no difference in expression among all groups. The protein expression level of MMP-12 in the lens injury combined with optic nerve injury group was up-regulated compared with the control group and optic nerve injury group at 7, 14 and 21d after successful modeling(P<0.05), and it was up-regulated in the lens injury group combined with optic nerve injury group compared with optic nerve injury group at 21d(P<0.05). At 28d, there was no difference in expression among all groups.CONCLUSION: The up-regulated expression of MMP-12 may be involved in the long-term regeneration of the optic nerve after lens injury.
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Objective:To explore the role of matrix metalloproteinase 12 (MMP12) in airway macrophages and pulmonary neu-rogenic substance P ( SP ) in the pathogenesis of asthma by analyzing their relationship in different categories of asthmatic patients.Methods:Twenty patients of asthma remission phase ( remission asthma group ) , twenty ones of mild acute exacerbation asthma (mild asthma group) and twenty healthy adults (normal control group) were included,respectively.After lung function was measured,the numbers of macrophage in induced sputum were counted.The expression levels of MMP12 mRNA and protein in sputum macrophages were detected by quantitative reverse transcription polymerase chain reaction and Western blot.The concentration of sputum SP was assayed by enzyme immunometric assay.Results: ( 1 ) Compared with the subjects in normal control group, forced expiratory volume in 1 second%predicted ( FEV1 ) and forced expiratory flow rates at 50% of the forced vital capacity % predicted ( FEF50 ) were much lower and the numbers of sputum macrophages were much higher in the patients in different asthmatic groups.Compared with the patients in remission asthma group,FEV1 and FEF50 were much lower in the ones in mild asthma group.(2) MMP12 expressions in the macrophages and the concentrations of SP in sputum were significantly increased in the patients in different asthmatic groups compared with those in normal control group;Furthermore,MMP12 and SP in mild asthma group were much higher than in remission asthma.(3) In all patients from different asthmatic groups,mRNA expressions of MMP12 in the macrophages were positively correlated with the levels of sputum SP or the numbers of sputum macrophages,whereas negative correlations between mRNA expressions of MMP 12 and FEV 1 or FEF50 were observed.Conclusion: The regulatory imbalance of macrophages′MMP12 and pulmonary neurogic SP may participate in the pathogenesis of asthma and become the potential targets for asthma therapy.
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Objective To investigate the roles of interleuldn (IL)-17,IL-10,and matrix metalloproteinase12 (MMP-12) in the pathogenesis of atherosclerosis and further to understand the intervention mechanism of atorvastatin for atherosclerosis.Methods Eighteen Japanese white rabbits were randomly divided into 3 groups:control,model and intervention groups (n =6 in each group).The control group,model group and intervention group were fed basic diet,high-fat diet,and high-fat diet plus atorvastatin (5 mg/(kg·d),respectively for 12 weeks.The levels of serum total cholesterol (TC) and triglyceride (TG) were determined by enzymatic colorimetric method; the levels of low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were determined by selective precipitation method; the levels of IL-17,IL-10,and MMP-12 were measured by double antibody sandwich enzyme-linked immunosorbent assay (ELISA); the pathological morphology changes of atherosclerotic plaque was observe by HE staining; and the thickness of the intima,intima/media ratio,plaque area,and plaque proportion were calculated by image analysis software.Results The expression levels of serum TC,TG,LDL-C,and IL-17,as well as those of IL-17 and MMP-12 of the intervention group were significantly lower than those of the model group (all P ＜0.05),but they were still higher than the control group (all P ＜0.05),while there was no significant difference in the level of HDL-C.The level of serum IL-10 of the intervention group was higher than that of the model group and the control group (all P＜0.05).The thickness of intima,intima/media ratio,plaque area,and plaque proportion of the intervention group were all significantly lower than those of the model group (all P ＜0.05).Correlation analysis showed that the plaque proportion was significantly positively correlated with IL-17 (r =0.847,P =0.001) and MMP-12 (r =0.839,P =0.001),and was negatively correlated with IL-10 (r =0.794,P =0.002); IL-17 was significantly positively correlated with MMP-12 (r =0.709,P=0.001),and was significantly negatively correlated with IL-10 (r =0.738,P＜0.001).MMP-12 was significantly negatively correlated with IL-10 (r =0.563,P =0.015).Conclusion The increased levels of IL-17 and MMP-12,as well as the decreased level of IL-10 illustrate that inflammatory factors play an important pathological role in the pathogenesis of atherosclerosis.In addition to lipid-lowering effect,atorvastatin may decrease the degree of chronic arterial inflammatory response and play a role in anti-atherosclerosis by decreasing the expression of IL-17 and MMP-12,as well as increasing the expression of IL-10.
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BACKGROUND: Gelsolin and matrix metalloproteinase 12 (MMP12) expression has been reported in Langerhans cell histiocytosis (LCH), but the clinical significance of this expression is unknown. We investigated the associations of these proteins with clinical manifestations in patients diagnosed with LCH. METHODS: We performed a retrospective analysis of clinical data from patients diagnosed with LCH and followed up between 1998 and 2008. Available formalin-fixed, paraffin-embedded specimens were used for gelsolin and MMP12 immunohistochemical staining. We analyzed the expression levels of these proteins and their associations with LCH clinical features. RESULTS: Specimens from 36 patients (20 males, 16 females) with a diagnosis of LCH based on CD1a positivity with clinical manifestations were available for immunohistochemical staining. Median patient age was 62 months (range, 5 to 207). The expression of gelsolin varied; it was high in 17 patients (47.2%), low in 11 patients (30.6%), and absent in 8 patients (22.2%). The high gelsolin expression group had a higher tendency for multi-organ and risk organ involvement, although the trend was not statistically significant. MMP12 was detected only in 7 patients (19.4%) who showed multi-system involvement (P=0.018) and lower event-free survival (P=0.002) in comparison to patients with negative MMP12 staining. CONCLUSION: Gelsolin and MMP12 expression may be associated with the clinical course of LCH, and MMP12 expression may be particularly associated with severe LCH. Further studies of larger populations are needed to define the precise role and significance of gelsolin and MMP12 in the pathogenesis of LCH.
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Humans , Male , Disease-Free Survival , Gelsolin , Histiocytosis , Histiocytosis, Langerhans-Cell , Immunohistochemistry , Langerhans Cells , Matrix Metalloproteinase 12 , Proteins , Retrospective StudiesABSTRACT
Objective To investigate the association of serum levels of matrix metalloproteinase-12 (MMP-12) and a functional polymorphism in the promoter - 82A/G of the MMP-12 gene with acute atherosclerotic cerebral infarction (ACI).Methods All 608 cascs of acute ACI and 374 healthy controls were included in the study.Serum levels of MMP-12 were measured using the enzyme-linked immunosorbent assay (ELISA). At the same time,polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed on the - 82A/G polymorphism in the MMP-12 gene.Results The serum levels of MMP-12 in ACI group (( 17.36 ± 9.12) ng/ml) were not significantly higher than those in the healthy control group ( ( 17.42 ± 7.70) ng/ml,t ＝ 0.047,P =0.962).The frequency of the AG + GG genotypes was not significantly different between the two groups (7.6％ vs 5.9％,x2 =0.281,P =0.584),and the frequencies of the G allele were 3.8％ and 2.9％ in the ACI group and the control group respectively which were also not significantly different between the two groups ( x2 =0.746,P =0.374).Conclusion There is no correlation between serum level with genetic polymorphism in MMP-12 promoter (-82A/G) and cerebral infarction.
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Objective This study was aimed to determine the expression of matrix metalloproteinase-11 ( MMP-11 ) and MMP-12 in cervical cancer and the relationship between their expression with clinicopathological characteristics and the potential use as a prognostic and therapeutic agent.Methods Streptavidin-biotin-protein-linked peroxidase method(ie,SP) immunohistochemical method was used to detect the expression of MMP-11,MMP-12 in 20 cases of normal cervical tissue,36 cases of CIN tissue,g0 cases of cervical cancer.MMP-11 and 12 expression in cervical cancer and clinicopathological parameters of relevance were analyzed,and the relationship between MMP-1 1,-12 was studied in the development of cervical cancer for a preliminary study.Results The positive expression rate (82.50％,g1.25％ )of MMP-11 and-12 in cervical cancer was significantly higher than that of cervical dysplasia ( CIN ) specimens ( 69.44％,61.11％ ) and normal cervical tissues(5.00％,0),the difference was statistically significant ( P ＜0.05).(2)The expression of MMP-11 and-12 in cervical histological grade,clinical depth of stromal infiltration,with or without vascular invasion,with or without lymph stage,node metastasis was different.The difference was statistically significant ( P ＜0.05).(3)MMP-11 and-12 expression was significantly correlated.Conclusions The expression of MMP-11and-12 in cervical cancer tissues prompts that MMP-11 and-12 plays a central role in the course of invasion and metastasis of cervical carcinoma.MMP-11 and-12 joint detection may have a reference value in terms of the inchoate diagnosis of the cervical cancer,the evaluation and prognosis of tumor malignances degree.
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Objective To investigate the influences of lysozyme on the mRNA expressions of MMP-1,-12 and lysyl oxidase(LOX)in cultured fibroblasts in vitro.Methods Primarily cultured fibroblasts isolated from human skin were treated with three concentrations(0.1×10~(-8),1×10~(-7)mol/L)of lysozyme followed by another 24-hour cuhure.Subsequently,total RNA was extracted from the fibroblasts and subjected to RT-PCR for the detection of MMP-1,-12 and LOX mRNA.Results There was a significant difference in the mRNA expressions of MMP-1,-12 and LOX among the fibroblasts treated with the three concentrations of lysozyme (F=6.98,4.44,5.24,respectively,all P<0.05).SNK-q test showed that untreated fibroblasts differed signifi-cantly from those treated with lysozyme of 1×10~(-7) mol/L in the mRNA expression of MMP-1 and MMP-12 (P<0.05),and from those treated with iysozyme of 1×10~(-7) mol/L.and 1×10~(-8)mol/L in the mRNA expres-sion of LOX(both P<0.05),whereas no significant difference was ohserved between fibroblasts treated with lysozyme of 1×10~(-8) mol/L and untreated fibrohlasts or those with lysozyme of 1 x 10~(-7)mol/L in the mRNA expression of MMP-1 and MMP-12.or between fibroblasts treated with lysozyme of 1 x 10~(-8)mol/L and those with that of 1×10~(-7) mol/L in the expression of LOX (all P>0.05).Conclusions Lysozyme upregulates the mRNA expression of MMP-1 and MMP-12 but downregulates the mRNA expression of LOX in cultured fibro-blasts in vitro.
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Objective To observe the effects of over-expression of matrix metalloproteinase-12 (MMP-12) on the transformation of pulmonary fibroblasts in radiation damaged rats. Methods Thirty-two male Wistar rats (weighed 250-280g) were randomly assigned into control group and 1, 2 and 4 weeks after irradiation groups (8 each). The whole lungs of rats in irradiation groups were irradiated by 60 Co ?-ray at a dose of 20Gy, and the lung specimens were harvested 1, 2 and 4 weeks after radiation. The change of MMP-12 activity was detected by gelatin zymography, the degradation and collapse of elastic fibers were observed by tissue specific staining, the "cross talking" phenomenon between alveolar type Ⅱ cells and mesenchymal cells was observed by transmission electron microscopy, the content of TGF-?1 was determined by ELISA, and the expression of ?-smooth muscle actin (?-SMA) was examined by immunohistochemistry. Results MMP-12 activity began increasing 1 week after irradiation, and seemed to decrease 4 weeks after irradiation. Elastin, a part of the basement membrane of alveolar wall, began to degrade and collapse 1 week after radiation, and became worse 4 weeks after irradiation. The expressions of both TGF-?1 and ?-SMA were elevated gradually within 4 weeks after radiation. The "cross talking" phenomenon was found by electron microscopy between alveolar type Ⅱ cells and mesenchymal cells. Conclusions Increased activity of pulmonary MMP-12 has been found after radiation, which may promote the transformation of pulmonary fibroblasts by degrading elastin and ultimately initiate the pulmonary fibrosis.