Role of extracellular matrix metalloproteinases in cardiac remodelling

The latent collagenolytic system is an intrinsic part of normal myocardium. Controlled activation of this system becomes necessary in ventricular chamber remodeling following inflammation and injury, such as dilated cardiomyopathy and myocardial infarction. Evidence exists to indicate activation of collagenolytic enzymes in patients with congestive heart failure and dilated, dysfunctional ventricles due to cardiomyopathy and ischemic heart disease.     

基质金属蛋白酶及其组织抑制剂与病毒性心肌炎心肌纤维化

心肌纤维化是病毒性心肌炎向扩张型心肌病转化的重要环节,由心肌胶原代谢失衡引起,近年来基质金属蛋白酶及其组织抑制因子(MMPs/TIMPs)这一胶原降解系统比例失调造成病毒性心肌炎中胶原合成降解代谢紊乱并导致心肌发生纤维化。     

Matrix metalloproteinases and their inhibitors in gastric cancer

Background—The matrix metalloproteinases (MMPs)and tissue inhibitors of matrix metalloproteinases (TIMPs) are stronglyimplicated in tumour invasion and metastasis.
Aims—To investigate the presence of individualMMPs and TIMPs in gastric cancer.
Methods—The presence of MMP-1, MMP-2, MMP-3,MMP-9, TIMP-1, and TIMP-2 was identified in a group of gastric cancers(n=74) by immunohistochemistry using monoclonal antibodies. Theseantibodies were effective on formalin fixed, paraffin wax embedded sections.
Results—A large proportion (94%) of gastriccancers contained MMP-2; MMP-1 and MMP-9 were also detected in 73% and70% of tumours respectively. MMP-3 was only present in 27% oftumours. MMP-1 and MMP-9 were found predominantly in intestinal typetumours. TIMP-1 and TIMP-2 were identified in 41% and 57% of tumoursrespectively. Immunoreactivity for individual MMPs or TIMPs was notidentified in normal stomach.
Conclusions—This study shows the presenceof matrix metalloproteinases, particularly MMP-2, and TIMPs in stomachcancer. Antibodies which are effective in formalin fixed, paraffin waxembedded sections are useful for the identification of MMPs and TIMPsin diagnostic specimens.

Keywords:immunohistochemistry; matrix metalloproteinase; neoplasm; stomach

     

乙型肝炎病毒X蛋白对MMPs及TIMPs的影响

目的全面分析乙型肝炎病毒X蛋白（Hepatitis Bvirus Xprotein，HBx）对基质金属蛋白酶（Matrix Metalloproteinases，MMPs）及组织金属蛋白酶抑制物（Tissue Inhibitors of Metalloproteinases，TIMPs）的影响，探讨其在肝细胞癌的侵袭转移中的可能作用。方法PCR扩增HBVX基因并克隆人真核表达载体pcDNA3．1／HisC，重组载体及空载体分别以Lipofectamine2000转染HepG2细胞并以800μg／mlG418筛选抗性细胞克隆。以Western blot检测抗性细胞HBx表达。抽提细胞总RNA，半定量RT-PCR检测MMPs及TIMPs。收集细胞培养液上清，以明胶酶谱检测MMP2及MMP9活性，反相明胶酶谱检测TIMPs活性。结果构建了HBx重组载体pcDNA3．1-XB。该重组载体及对照空载体转染HepG2细胞后，G418筛选，分别获得抗性细胞克隆HepG2-XB及HepG2-HIS，前者经Westernblot证实可表达HBx。半定量RT—PCR显示HBx可促进MMP2、7、13、14、16、17、19、23、24及TIMP1、4基因的转录，抑制MMP1、3、8、9、10、11、12、15、20及TIMP2、3基因的转录。明胶酶谱检测显示HBx可促进酶原MMP2（Pro—MMP2）及活性MMP9（Active—MMP9）表达，抑制酶原MMP9（Pro—MMP9）表达；反相明胶酶谱显示HBx可促进TIMP1、TIMP4的表达，同时抑制TIMP2及糖基化TIMP3的表达。结论HBx蛋白对MMPs、TIMPs转录表达的影响是多方面的，但这种影响在HBx促进肝癌细胞侵袭转移过程中的确切机制有待进一步阐明。     

基质金属蛋白酶及组织抑制剂在肝癌中的表达及与预后的关系

目的：探讨基质金属蛋白酶（MMPs)及其组织抑制剂（TIMPs）在肝癌中的表达及其意义。方法：应用免疫组化、Northern印迹杂交及图像分析技术对人肝细胞癌（HCC）、肝癌细胞株7721、全反式维甲酸处理的7721细胞（RA－7721）和正常人肝细胞株L－02作MMP－1、2、9和TIMP－2的表达分析。结果;肝癌细胞浆内可表达MMP－1、2和9,但癌内阴性组5年生存率明显高于相应的阳性组（P＜0．05）。体外MMP－9mRNA在7721细胞表达明显高于RA－7721以及L－02细胞,而TIMP－2mRNA的表达与MMP－9相反,MMP－2mRNA在7721细胞中的表达仅略高于L－02细胞。结论：HCC组织内MMP－1和9的表达与患者的预后密切相关;癌细胞高表达MMP－9、低表达TIMP－2,可能是肝癌细胞浸润、转移的主要基础,而MMP-2并无重要作用。     

胃癌组织中CTGF蛋白表达及其临床意义

目的探讨结缔组织生长因子（CTGF）蛋自在胃癌发生、发展中的作用。方法采用免疫组化法检测10份正常胃黏膜组织、50份胃癌组织及其癌旁组织中CTGF蛋白表达情况。结果胃癌组织中CTGF阳性表达高于癌旁组织及正常胃黏膜组织,P〈0．05;癌组织CTGF阳性表达率明显高于正常黏膜组织（P〈0．01）,CTGF的高表达与胃癌分化程度、淋巴结转移密切相关（P均〈0．05）;与肿瘤浸润的深度无明显关系。结论CTGF蛋白可作为胃癌前病变及胃癌早期诊断和预后判断的指标。     

Role of melatonin in regulating matrix metalloproteinase-9 via tissue inhibitors of metalloproteinase-1 during protection against endometriosis

Abstract:  Endometriosis is a gynecological disease of women and plausibly regulated by matrix metalloproteinases (MMPs). However, mechanisms of alterations in MMPs during endometriosis remain unclear. Human endometriotic tissues possessing varying degrees of severity were examined for expression of MMPs and tissue inhibitors of metalloproteinase (TIMP)-1. In addition, endometriosis was generated in mice and endometriotic tissues were tested for MMP-9 activity. Results show significant upregulation of secreted and synthesized proMMP-9 activity with duration and severity of endometriosis. Along with upregulation of activity, the expression of proMMP-9 was found increased while TIMP-1 expression followed an inverse trend. The effect of melatonin, a major secretory product of the pineal gland, on endometriosis was examined in preventive and therapeutic models in mice. The results show that melatonin arrested lipid peroxidation and protein oxidation and downregulated proMMP-9 activity and expression in a time and dose-dependent manner while protecting and regressing peritoneal endometriosis. Moreover, the attenuated activity and expression of proMMP-9 were associated with subsequent elevation in the expression of TIMP-1. Our study reveals for the first time the role of melatonin in arresting peritoneal endometriosis in mice and a novel marker, expression ratio of proMMP-9 versus TIMP-1, was identified for assessing severity and progression of endometriosis.     

Prognostic significance of p53 and matrix metalloproteinase-9 expression in follicular lymphoma

Objectives: p53 mutations and high protein expression are associated with adverse prognosis in several lymphoma subtypes. Matrix metalloproteinase‐9 (MMP‐9) has also been found to correlate with poor survival in all lymphomas studied. The data concerning the clinical role of protein expression of p53 or gelatinases and their inhibitors in follicular lymphoma are rare. The purpose of this study was to evaluate the prognostic and clinical implications of the immunoreactive proteins p53, MMP‐2, MMP‐9, tissue inhibitor of matrix metalloproteinase‐1 (TIMP‐1) and TIMP‐2 in follicular lymphoma. Methods: The material consisted of 67 patients with primarily non‐transformed follicular lymphoma. Diagnostic lymph node tissue sections of patients were stained by immunohistochemical method using specific monoclonal antibodies. Results: p53 over‐expression was detected in 8 (12%) out of 67 cases. p53 over‐expression correlated with high grade (P = 0.011), bulky tumour (P = 0.031) and forthcoming transformation (P = 0.001). It also correlated with poor overall (P = 0.001) and cause‐specific survival (P = 0.010) in multivariate analysis and had a strong inverse correlation with time to transformation (P < 0.001). MMP‐2, MMP‐9 and TIMP‐2 expression correlated with high grade. MMP‐9 positivity in centroblasts correlated with good chemotherapy response (P = 0.019), but it was not prognostic for survival. MMP‐2, TIMP‐1 or TIMP‐2 did not associate with survival, either. Conclusions: In this study, p53 over‐expression predicted both transformation to diffuse large B‐cell lymphoma and poorer overall and cause‐specific survival of patients with follicular lymphoma. Expression of gelatinases or their inhibitors did not have any significant correlations with prognosis, although MMP‐9 predicted a good response to first‐line chemotherapy.     

Simultaneous up-regulation of matrix metalloproteinases 1, 2, 3, 7, 8, 9 and tissue inhibitors of metalloproteinases 1, 4 in serum of patients with chronic obstructive pulmonary disease

Background and objective: Matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP) have been implicated in chronic obstructive pulmonary disease (COPD) pathogenesis. However, the majority of studies have focused on single MMP, and there is limited information on parallel expression of MMP and their antagonists TIMP. We, therefore, investigated the serum profile of MMP 1–3, 7–9, 12 and 13, and TIMP 1–4 in COPD patients. Methods: Serum MMP 1–3, 7–9, 12 and 13, and TIMP 1–4 were measured in 74 COPD patients and 20 control subjects by multiple microsphere technology. Results: MMP 1–3 and MMP 7–9 were elevated in COPD patients compared with control subjects (P= 0.001–0.043). The increased concentrations of MMP 1, 8 and 9 paralleled GOLD stage (P= 0.002–0.007). TIMP 1 and 4 concentrations were elevated in COPD (P < 0.001). MMP 1, 8 and 9, and TIMP 1 and 4 serum levels in COPD non‐smokers were higher than in control non‐smokers (P= 0.002–0.025). MMP 12 and 13 levels were undetectable in our serum samples. Conclusions: This study provides further evidence for increased MMP 1, 7–9, and TIMP 1 serum levels in COPD, and demonstrates for the first time serum elevation of MMP 2 and 3, and TIMP 4. The finding that circulating TIMP 4 levels are increased in COPD and the observed relationship between serum levels of MMP 1, 8 and 9, and GOLD stage requires verification in an expanded patient cohort.     

Matrix metalloproteinase-9 activity and expression is reduced by melatonin during prevention of ethanol-induced gastric ulcer in mice

Matrix metalloproteinases (MMPs) play an important role in degradation of gastric extracellular matrix proteins. However, no reports are available on the relationship between the activity of MMPs and gastric ulceration induced by alcohol. Our objective was to investigate the effect of melatonin (N-acetyl-5-methoxytryptamine) on the regulation of MMP-9 and MMP-2 activities during prevention of ethanol-induced gastric ulcer. Biochemical and zymographic methods were used to analyze MMP-9 and -2 activities in gastric tissues of Balb/c mice following induction of gastric ulcer by ethanol. Our studies reveal that melatonin arrested cell injury, protein carbonyl formation, and lipid peroxidation in mice during gastroprotection. Melatonin dose-dependently reduced proMMP-9 activity that was induced ( approximately 25-fold) during ethanol-induced gastric ulceration. Severity of gastric ulcers were correlated proportionately with increased dose of ethanol and elevated activity of proMMP-9 and -2. The reduced activities of MMP-9 and -2 were associated with reduced expression of TNF-alpha and increased expression of tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2). We conclude that melatonin's ability to prevent ethanol-induced gastric ulceration in mice is related to a reduction in proMMP-9 activity and expression.     

Serum levels of matrix metalloproteinase‐9, tissue inhibitors of metalloproteinase‐1 and their ratio are associated with impaired lung function in the elderly: A population‐based study

Background and objective: Matrix metalloproteinases (MMP) and their inhibitors, tissue inhibitors of metalloproteinases (TIMP), regulate homeostasis and turnover of the extra cellular matrix. The aim of this study was to investigate the associations of serum MMP‐9 and TIMP‐1 with lung function. Methods: Spirometry was performed in a population‐based sample of 888 subjects aged 70 years. Serum MMP‐9 and TIMP‐1 concentrations were measured by ELISA. Results: Lower FEV₁ values were associated with higher serum levels of MMP‐9 (P = 0.001) and TIMP‐1 (P < 0.001), and a higher ratio of MMP‐9 to TIMP‐1 (P = 0.02). These associations were significant after adjustment for gender, weight, height, BMI, current smoking, pack years of smoking and the time for which samples were frozen. After stratification for gender, the associations between FEV₁ and MMP‐9, TIMP‐1, and their ratio, were significant in men but not in women. Conclusions: Lower FEV₁ was significantly but weakly associated with higher serum levels of MMP‐9, TIMP‐1 and a higher MMP‐9/TIMP‐1 ratio. This association was stronger in men than in women, suggesting a possible role for extracellular matrix remodelling in the development of impaired lung function. These associations may also partly explain the association between low FEV₁ and cardiovascular disease.     

辛伐他汀对动脉粥样硬化斑块中MMPs及TIMPs表达的影响

目的 :观察辛伐他汀对动脉粥样硬化斑块中基质金属蛋白酶 （MMPs）及其组织抑制因子 （TIMPs）表达的影响。方法 :新西兰大白兔 48只随机分为试药组和对照组 ,用 PTCA球囊导管拉伤腹主动脉 ,高脂饮食喂养 4周后 ,试药组 （2 4只 ）给予辛伐他汀 5m g· kg- 1· d- 1,对照组 （2 4只 ）以淀粉为对照 ;每组按实验终点 （给药后 2、4和 8周 ）随机分为 3个亚组 ,继续高脂饮食喂养至实验结束。动物高脂喂养前、每周和处死前取静脉血 2 ml检测血脂变化 ,取拉伤段动脉用于总 RNA提取和组织病理检测。结果 :高脂饮食喂养 1周后 ,2组动物血脂水平在正常的 5～10倍 ,组间无显著差异 （P>0 .0 5） ;用药后 2周 ,试药组血脂明显低于对照组 ;用药后 4周 ,试药组血管组织MMP1,2低于对照组 ,TIMP1/ MMP1明显大于对照组 （P <0 .0 5,P<0 .0 1） ,TIMPs表达两组无显著差异 （P >0 .0 5）。结论 :辛伐他汀通过选择性地改变局部 MMPs和 TIMPs表达 ,减少基质分解破坏 ,增加局部斑块的稳定性     

Marrow matrix metalloproteinases (MMPs) and tissue inhibitors of MMP in acute leukaemia: potential role of MMP-9 as a surrogate marker to monitor leukaemic status in patients with acute myelogenous leukaemia

Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) were demonstrated to have important implications in the progression and invasiveness of many malignant disorders. In contrast, the biological significance of these molecules in human leukaemias is not clear. We determined the levels of MMP-2, MMP-9, TIMP-1 and TIMP-2 in the bone marrow of 37 patients with acute myelogenous leukaemia (AML) and 18 patients with acute lymphoblastic leukaemia (ALL) before chemotherapy. Nineteen bone marrow donors served as normal controls. After chemotherapy, sequential measurements were done during the course in 19 AML patients. The levels of TIMP-1 and TIMP-2 were significantly higher and MMP-9 levels were significantly lower in the AML and ALL patients than in the normal controls. MMP-2 levels were higher in ALL, but not AML patients, compared with controls. Moreover, the levels of marrow MMP-2 and MMP-9 did not parallel the numbers of leukaemic blasts in the peripheral blood. MMP-9 levels were significantly lower in the AML patients who achieved a complete remission (CR) than in those who did not (8.71 +/- 8.15 ng/ml vs 26.13 +/- 27.75 ng/ml, P < 0.05). The AML patients with lower MMP-9 levels (< or = 4.4 ng/ml) tended to have longer survival time than those with higher levels (> 12 months vs 4 months, P = 0.12). In addition, MMP-9 levels in the AML patients at CR rose to the same range as the controls, but dropped again at relapse, demonstrating a close relationship of marrow MMP-9 with disease status of AML. Therefore, we conclude that the level of marrow MMP-9 may be a useful surrogate marker for monitoring disease status in AML and propose it as a potential prognostic factor.     

Plasma matrix metalloproteinase and tissue inhibitor of metalloproteinase in patients with agnogenic myeloid metaplasia or idiopathic primary myelofibrosis

Agnogenic myeloid metaplasia (AMM) is characterized by bone marrow fibrosis with abnormal accumulation of extracellular matrix components (ECM), which is dependent on the balance between matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). Twenty-five patients with AMM, 30 with essential thrombocythemia (ET), 12 with polycythemia vera (PV) and 20 normal control subjects were studied. AMM patients had decreased plasma levels of MMP-3 and marked elevated levels of TIMP-1, but MMP-1, MMP-2 and MMP-9 levels were not significantly different from control subjects. Elevated levels of plasma TIMP-1, but not MMPs, were found in ET and PV. Reduced MMP activity together with increased TIMP-1 activity may be essential in fibrosis formation.     

女性压力性尿失禁患者阴道壁组织中MMP-13、MMP-14和TIMP-1的表达变化及意义

目的探讨女性压力性尿失禁（SUI）患者阴道壁组织中基质金属蛋白酶13、14（MMP-13、14）和组织基质金属蛋白酶抑制剂-1（TIMP-1）的表达变化及意义。方法收集30例女性SUI患者的阴道壁组织（SUI组）、30例盆腔器官脱垂患者（POP）的阴道壁组织（POP组）、30例无SUI及POP患者的阴道壁组织（对照组）,采用免疫组化SP法检测这三组的MMP-13、MMP-14、TIMP-1。结果MMP-13、MMP-14在POP组、SUI组中的表达明显高于对照组（P均〈0.05）,TIMP-1在POP组、SUI组的表达明显低于对照组（P均〈0.05）;SUI组MMP-13的表达与TIMP-1表达呈负相关（r=-0.281,P〈0.05）,MMP-14与TIMP-1的表达呈负相关（r=-0.381,P〈0.05）,MMP-13与MMP-14的表达呈正相关（r=0.992,P〈0.05）。结论女性SUI患者阴道壁组织中MMP-13、14表达增加,TIMP-1表达减少,导致胶原降解增加,从而诱发SUI。     

Serum levels of tissue inhibitor of metalloproteinases-2 and of precursor form of matrix metalloproteinase-2 in patients with liver disease

Abstract: Serum levels of tissue inhibitor of metalloproteinases-2 (TIMP2) and of precursor form of matrix metalloproteinase-2 (proMMP2) were determined in patients with chronic hepatitis and hepatocellular carcinoma by a one-step sandwich enzyme immunoassay. Serum levels of TIMP2 and proMMP2 were significantly higher in patients with chronic liver disease, than in normal controls. Serum levels of TIMP2 showed a weak negative correlation with the serum albumin level and prothrombin time (PT). Serum levels of proMMP2 in patients with chronic hepatitis were strongly correlated with those of type IV collagen and were negatively correlated with PT and serum albumin levels. Serum proMMP2 levels were also significantly correlated with histological stages. These data indicate that serum levels of proMMP2 might be useful in the follow-up of patients with chronic hepatitis.     

Extracellular matrix remodelling in human aortic valve disease: the role of matrix metalloproteinases and their tissue inhibitors.

AIMS: Aortic valve diseases are characterized by pathological remodelling of valvular tissue but the cellular and molecular effectors involved in these processes are not well known. The role of matrix metalloproteinase (MMP)-2, MMP-9, MMP-3, MMP-7, and tissue inhibitor of matrix metalloproteinase (TIMP)-1 and TIMP-2 are investigated here. METHODS AND RESULTS: Histological analysis of pathological valves [aortic stenosis (AS) (n=49), aortic regurgitation (AR) (n=23)] and control valves (n=8) was performed. The main tissue abnormalities (calcification, inflammatory cells, and capillaries) observed in AS were less severe or absent in AR. However, both groups of pathological valves displayed similar histological signs of extracellular matrix (ECM) remodelling. Biochemical analysis of MMPs and TIMPs (gelatin and casein zymography and ELISA) was performed on valve extracts. MMP-2 activity was not significantly different in control and pathological valves. Increases in MMP-9 and MMP-3 in AS demonstrated an inflammatory state. Finally, there was a four- to seven-fold increase of TIMP-1 in pathological valves. TIMP-1, TIMP-2, and MMP-2 were synthesized by the valvular interstitial cells in primary culture. CONCLUSION: This study demonstrates the involvement of the MMP/TIMP system in ECM remodelling of both AS and AR. These findings provide evidence of inflammatory injury more severe in AS than in AR and involvement of mesenchymal cell response.     

The matrix metalloproteinases and their inhibitors in pancreatic cancer

Summary The matrix metalloproteinases (MMPs) and their tissue-specific inhibitors (TIMPs) are described and their roles in tumor invasion and metastasis are reviewed. The expression and activity of the MMPs and TIMPs in pancreatic cancer is reported and illustrated with immunohistochemistry andin situ hybridization. The role of MMP inhibitors (MMPIs) is reviewed in vivo and the use of novel MMPIs, e.g., BB94 (Batimastat) and BB2516 (Marimastat), in animal experiments are also described. Finally, the preliminary results from a phase 2 trial of BB2516 (Marimastat) in pancreatic cancer are reported.