慢性肝病患者血清TIMP-1和TIMP-2的变化及其临床意义

目的探讨慢性乙型肝炎(CHB)和乙型肝炎后肝硬化(LC)患者血清中的基质金属蛋白酶-1组织抑制因子(TIMP-1)、基质金属蛋白酶-2组织抑制因子(TIMP-2)的变化及其临床意义。方法用酶联免疫吸附试验(ELISA)检测75例CHB患者和41例乙型肝炎后LC患者血清TIMP-1、TIMP-2含量,并与30名健康献血员比较。结果CHB轻度、中度、重度和LC A级、B级、C级患者血清TIMP-1均明显高于正常对照组(P均<0.01),与透明质酸(HA)、层黏连蛋白(LN)、Ⅳ型胶原(CⅣ)呈正相关(r=0.935、0.836、0.910,P均<0.01);CHB重度和LC A级、B级、C级患者血清TIMP-2均明显高于正常对照组(P均<0.01),与HA、LN、CⅣ呈正相关(r=0.765、0.623、0.716,P均<0.01)。结论血清TIMP-1、TIMP-2在慢性肝病患者肝纤维化过程中明显升高,并与肝纤维化指标呈正显著性相关,可作为判断肝纤维化程度的指标。     

慢性肝病患者血清基质金属蛋白酶-2及其抑制物的变化和临床意义

目的探讨慢性乙型肝炎（CHB）和乙型肝炎后肝硬化患者血清中的基质金属蛋白酶（MMP-2）、基质金属蛋白酶组织抑制因子（TIMP-2）的变化及其临床意义。方法用ELISA法检测75例CHB患者、41例乙型肝炎后肝硬化患者和30例健康献血员血清MMP-2和TIMP-2含量，用放射免疫法检测透明质酸（HA）、层粘连蛋白（LN）和Ⅳ型胶原（CⅣ）的含量，分析MMP-2、TIMP-2与肝纤维化指标之间的相关性。结果CHB中度、CHB重度、肝硬化A级、B级、C级患者血清MMP-2均明显高于正常对照组（P〈0．01），且肝硬化C级〉肝硬化B级〉肝硬化A级〉CHB重度〉CHB中度〉CHB轻度（P〈0．05，0．05，0．05，0．01，0．01），与HA、LN、C Ⅳ呈显著性正相关（r=0．913，0．845，0．897。P均〈0．01）；CHB重度、肝硬化A级、B级、C级患者血清TIMP-2均明显高于正常对照组（P〈0．01），肝硬化C级与CHB轻度、CHB中度、CHB重度比有显著性差异（P〈0．01，0．01，0．05），肝硬化B级、肝硬化A级、CHB重度与CHB轻度、CHB中度比有显著性差异（P〈0．01），与HA、LN、CⅣ呈显著性正相关（r=0．693，0．574，0．646。P均〈0．01）。结论血清MMP-2、TIMP-2在慢性肝病患者肝纤维化过程中明显升高，可做为判断肝纤维化程度的指标，尤其以MMP-2的诊断意义更大。     

慢性乙型肝炎患者血清TGF—β1水平与肝纤维化的关系

目的：观察慢性乙型肝炎（CHB）患者血清转化生长因子β-（TGF—β1）在肝纤维化不同阶段的表达情况，探讨TGF—β1与肝纤维化程度的关系，评价其对肝维化诊断的价值。方法：采用酶联免疫吸附法（ELASA）检测40例慢性乙型肝炎患者血清TGF—β1水平．放射免疫法（RIA）检测血清透明质酸（HA）、层粘连蛋白（LN）、Ⅲ型前胶原氨基端肽（PⅢNP）、Ⅳ型胶原（CⅣ）水平，40例患者全部进行肝组织活检，分析TGF-β1与肝组织纤维化程度分期和炎症活动度分级的关系以及与HA、LN、PⅢNP、CⅣ四项指标的相关性。结果；（1）慢性乙型肝炎患者TGF-β1水平明照高于正常对照组（P〈0．01）．且随肝纤维化程度的加重而升高（P〈0．01），TGF-β1变化趋势与HA、LN、PⅢNP、CⅣ均呈正相关（P〈0．01）。（2）按炎症活动度（G）分组，TGF-β1在G1～G4组水平明显高于G0组（P〈0．01），但组间两两比较无显著性差异（P〉0．05）。结论：血清TGF—β1表达水平与乙肝患者肝纤维化程度密切相关，且不受肝组织炎症程度的影响，在早期肝纤维化的敏感性高于HA、LN、PⅢNP、CⅣ，可作为早期肝纤维化的诊断指标。     

慢性病毒性肝炎患者血清TIMP-1、HA与肝纤维化的关系

目的:通过检测慢性病毒性肝炎患者血清基质金属蛋白酶抑制因子-1(TIMP-1)、透明质酸(HA)、型前胶原氮端肽(PNP)、型胶原(C)、层粘连蛋白(LN)水平,分析TIMP-1、HA与肝纤维化的关系,探讨二者对肝纤维化的诊断价值。方法:酶联免疫法(ELISA)检测40例慢性病毒性肝炎患者及12例健康体检者血清TIMP-1,放射免疫法(RIA)检测其血清HA、PNP、C、LN水平,同时对患者行肝穿刺活检,以病理诊断为金标准,分析各项血清学指标与肝纤维化分期及炎症分级的关系。结果:血清TIMP-1在肝纤维化早期即明显升高,且随肝纤维化程度的加重而逐渐升高(P<0.01);血清HA在肝纤维化中后期升高;二者均与肝纤维化分期成正相关(rs分别为0.878、0.828,P<0.01)。结论:血清TIMP-1能区分肝纤维化不同时期,适于动态观察;血清TIMP-1、HA可作为肝纤维化无创性诊断的指标。     

乙肝患者肝纤维化进程中sICAM-1表达的研究

目的探讨可溶性细胞黏附分子-1（sICAM-1）的含量在乙型肝炎患者纤维化病程中的表达及意义。方法选择80例处在非急性发作期的乙型肝炎患者进行肝组织穿刺检测,分成三组,选取30例健康体检者作为对照组。上述四组均空腹抽取静脉血,留取血清使用酶联免疫吸附法（ELISA）检测各组血清中的sICAM-1含量;同时,采用放射免疫分析法检测纤维化四项包括血清透明质酸（HA）、层粘连蛋白（LN）、Ⅲ型前妥胶原（PCⅢ）、Ⅳ型胶原蛋白（Ⅳ．C）含量。结果sICAM-1在乙肝患者的血清含量随着肝纤维化严重程度呈阶梯样上升,有显著性差异（P〈0．05）。各组血清sICAM-1含量与肝纤维化指标HA、LN、Ⅳ.C呈正相关（P〈0．01）。结论sICAM-1可作为判断肝纤维化的血清学指标,动态检测肝纤维化进程。而且,sICAM-1也可以为临床肝纤维化检测提供非创伤性的早期诊断依据。     

激活素A与肝纤维化指标的相关性探讨

目的探讨慢性乙型肝炎、肝硬化患者血清激活素A（ActivinA，ACTA）与血清肝纤维化指标-透明质酸（HA）、层粘连蛋白（LN）、Ⅲ型前胶原（PCⅢ）的关系。方法采用酶联免疫（ELISA）法检测108例慢性乙型肝炎、肝硬化患者和20例正常对照组血清ACTA水平，同时检测血清HA、LN、PCⅢ含量。其中35例做肝组织活检，进行HE染色。结果慢性乙肝轻、中、重度组及肝硬化组ACTA水平明显高于对照组（P〈0．01或P〈0．05）。AC-TA水平与血清HA、LN、PCⅢ水平呈正相关（P〈0．01或P〈0．05）。血清HA、LN、PCⅢ含量随肝组织纤维化程度加重而升高（P〈0．01或P〈0．05）。结论慢性乙型肝炎、肝硬化患者血清ACTA与肝纤维化血清标志及肝组织病理改变相一致。     

TIMP-1在肝纤维化组织中的表达和外周血浓度与肝病理分级的相关性研究

目的探讨慢性乙型肝炎、乙型肝炎后肝硬化患者肝组织及血清中基质金属蛋白酶抑制因子-1(tissue inhibitors of metalloproteinase,TIMP-1)与肝脏炎症活动程度及纤维化程度的相关性.方法同步收集28例慢性乙型肝炎、乙型肝炎后肝硬化患者血清及肝穿刺活组织标本,用免疫组化法测定肝纤维化组织中TIMP-1阳性率,用ELISA法检测血清中TIMP-1水平;另对47例同时期肝功能正常的健康体检者用ELISA法检测血清TIMP-1作为对照组.结果慢性乙型肝炎、乙型肝炎后肝硬化患者肝组织中TIMP-1的阳性表达与肝纤维化程度呈正相关(r=0.386,P=0.042),血清TIMP-1与肝纤维化程度呈正相关(r=0.646,P=0.000),血清中TIMP-1与肝脏炎症活动度分级呈正相关(r=0.622, P=0.000);血清TIMP-1水平与肝组织中的TIMP-1表达无显著相关性(P>0.05).结论肝活检免疫组化检测TIMP-1对反映肝纤维化程度有重要意义;血清TIMP-1含量能间接反映慢性活动性肝炎肝脏损伤程度,对于诊断及预后判断有重要参考作用.     

基质金属蛋白酶抑制剂-1在肝纤维化诊断中的临床应用

目的 评估血清基质金属蛋白酶抑制剂-1 (TIMP-1)在在肝纤维化诊断中的临床价值.方法 285例慢性乙型肝炎(CHB)患者,根据肝穿刺病理诊断结果分为无明显纤维化组(S0-S1)和明显纤维化组(S2-S4).将病理诊断结果作为金标准,使用ROC曲线评估TIMP-1诊断肝纤维化的敏感度、特异性和符合率,并和肝纤维化标记物HA、PCⅢ、CⅣ、LN比较其诊断性能,结果 对照组、S0-S1组、S2-S4组TIMP-1血清浓度分别为146,6±29.2、161.2±43.2、221.6±93.8 μg/L,对照组和S2-S4组及S0-S1组和S2-S4组间差异有统计学意义(t分别为5.32、4.98,P均小于0.01).TIMP-1区分慢性乙肝明显纤维化和轻度纤维化的ROC曲线下面积(AUC)为0.841(95％CI:0.762～0.920),TIMP-1在临界值为170.3 μg/L时,其敏感度、特异度分别为71.5％、82.4％,高于HA(75.6％、72.5％)、PCⅢ(70.5％、76.5％)、CⅣ(63.6％、78.4％)、LN(79.5％、64.7％).结论 血清TIMP-1诊断肝纤维化有较高的敏感度和特异性,血清TIMP-1可能是预测肝纤维化分期一个较好的指标.     

血清基质金属蛋白酶抑制因子-1诊断肝纤维化的临床价值

目的：比较慢性病毒性肝炎患者肝纤维化不同阶段血清基质金属蛋白酶抑制因子-1（TIMP-1）水平，探讨血清TIMP-1诊断肝纤维化的临床价值。方法：用酶免及放免法检测40例慢性病毒性肝炎患者和12例健康体检者血清TIMP-1、透明质酸（HA）、Ⅲ型前胶原氮端肽（PⅢNP）、Ⅳ型胶原（CⅣ）、层粘连蛋白（LN）水平。所有患者同时行肝脏穿刺活检，将五项血清学指标与肝纤维化分期（S）及炎症分级（G）进行相关性分析，采用ROC曲线确定血清TIMP-1诊断肝纤维化及早期肝硬化的截断值。结果：血清TIMP—1在肝纤维化早期即明显升高，各分期间差异有显著性，与S、G及其他四项血清学指标正相关，与S相关性最强。血清TIMP-1以258．28μg／L为截断值，诊断肝纤维化（S≥2）的灵敏度为0．966，特异度为0．909；以389．30μg／L为截断值，诊断早期肝硬化（S=4）的灵敏度为0．833，特异度为0．882。结论：血清TIMP-1能明确区分肝纤维化不同时期，可作为肝纤维化无创性诊断的新指标。     

慢性肝病患者血清MMP-9 TIMP-1的检测及临床意义

目的研究基质金属蛋白酶- 9( MMP- 9)、基质金属蛋白酶组织抑制因子- 1( TIMP- 1)在慢性肝病患者肝纤维化过程中的表达和动态变化.方法用酶标法检测 75例慢性乙型病毒性肝炎和 40例乙肝后肝硬化患者血清中 MMP- 9和 TIMP- 1的浓度.结果血清 MMP- 9和 TIMP- 1浓度在慢性肝病患者肝纤维化过程中逐渐升高,与正常对照组比较有显著性意义( P<0.01);其中轻度患者的 MMP- 9与中度、重度、肝硬化之间,肝硬化与中度、重度之间两两比较有显著性差异( P<0.05或 P<0.01);除了慢性乙肝重度患者的 TIMP- 1与肝硬化之间比较无意义外,其余各组之间两两比较均有显著性差异( P<0.01).结论血清 MMP- 9和 TIMP- 1在慢性肝病患者肝纤维化过程中可能起促进作用,能够被用来做为判断肝纤维化程度的指标.     

Cartilage degradation and invasion by rheumatoid synovial fibroblasts is inhibited by gene transfer of TIMP-1 and TIMP-3

Matrix metalloproteinases (MMPs) are believed to be pivotal enzymes in the invasion of articular cartilage by synovial tissue in rheumatoid arthritis (RA). Here, we investigated the effects of gene transfer of tissue inhibitors of metalloproteinases (TIMPs) on the invasiveness of RA synovial fibroblasts (RASF) in vitro and in vivo. Adenoviral vectors (Ad) were used for gene transfer. The effects of AdTIMP-1 and AdTIMP-3 gene transfer on matrix invasion were investigated in vitro in a transwell system. Cartilage invasion in vivo was studied in the SCID mouse co-implantation model for 60 days. In addition, the effects of AdTIMP-1 and AdTIMP-3 on cell proliferation were investigated. A significant reduction in invasiveness was demonstrated in vitro as well as in vivo in both the AdTIMP-1- and AdTIMP-3-transduced RASF compared with untransduced SF or SF that were transduced with control vectors. in vitro, the number of invading cells was reduced to 25% (P<0.001) in the AdTIMP-1-transduced cells and to 13% (P<0.0001) in the AdTIMP-3-transduced cells (% of untransduced cells). Cell proliferation was significantly inhibited by AdTIMP-3 and, less, by AdTIMP-1. In conclusion, overexpression of TIMP-1 and TIMP-3 by Ad gene transfer results in a marked reduction of the invasiveness of RASF in vitro and in the SCID mouse model. Apart from the inhibition of MMPs, a reduction in proliferation rate may contribute to this effect. These results suggest that overexpression of TIMPs, particularly TIMP-3 at the invasive front of pannus tissue, may provide a novel therapeutic strategy for inhibiting joint destruction in RA.     

Development of an enzyme-linked immunosorbent assay to measure total TIMP-1 (Free TIMP-1 and TIMP-1 in combination with matrix-metalloproteinases) and measurement of TIMP 1 and CRP in serum

A panel of six monoclonal antibodies (MAbs) was raised against purified human fibroblast tissue inhibitor of metalloproteinase-1 (TIMP-1) and characterised. All possible antibody pairs were tested for their suitability as capture and revealing antibodies in a two-site enzymelinked immunosorbent assay (ELISA) to measure total TIMP-1 (both free TIMP-1 and TIMP-1 together with matrix metalloproteinases (MMPs)). Using the best combination of MAbs the assay was optimised. The sensitivity of detection of the assay was 1.4 ng/ml, and inter- and intra-assay coefficients of variation were between 10.4–13.7% and 8.8–9.7%, respectively. Dilution series of human cerebrospinal and synovial fluids, plasma and sera paralleled those of the TIMP-1 standard curve indicating that the immunoreactivity detected in these samples was authentic TIMP-1. TIMP-2 shows no detectable cross reactivity in this assay confirming that this ELISA is specific for TIMP-1. The levels of total TIMP-1 and collagenase were measured in conditioned medium from A2058 human melanoma cells cultured in the absence or presence of human recombinant interleukin-1 (hrlL-l). Total TIMP-1 was also measured in serum samples with known C-reactive protein (CRP) (n = 100) and ₁ antichymotrypsin (ACT) (n = 52) concentrations; no correlation was found between TIMP-1 levels and either of these acute phase reactants although the levels of TIMP-1 were raised when compared to normal sera. This ELISA provides a rapid and convenient procedure for the quantitation of total TIMP-1 in human biological fluids and supernatants from cultured cell lines.     

MMP-9及TIMP-3在子宫内膜异位症中的表达及意义

[目的]探讨基质金属蛋白酶（MMP-9）、金属蛋白的组织抑制因子-3（TIMP-3）在子宫内膜异位症（EMs）患者异位内膜、在位内膜以及正常子宫内膜中的表达及意义.[方法]检测EMs患者的异位内膜（28例,A组）、在位内膜（20例,B组）和正常子宫内膜（22例,C组）中 MMP-9、TIMP-3的表达,并用银染法观察腺上皮基底膜的完整性.[结果]A组MMP-9的表达高于B组和C组,B组高于C组,且差异均有显著性（P〈0.05）.A组TIMP-3的表达低于B组和C组,B组低于C组,且差异均有显著性（P〈0.05）.腺上皮的基底膜银染结果显示：MMP-9的染色强度比值与基底膜的完整性呈负相关趋势（P〈0.01）,TIMP-3的染色强度比值与基底膜的完整性呈正相关趋势（P〈0.01）.在位内膜腺上皮基底膜完整率低于C组子宫内膜.[结论]①MMP-9的高表达和TIMP-3的低表达可能参与EMs的发病.②MMP-9与TIMP-3的平衡失调可能是EMs的发病原因之一.     

子宫内膜异位症中金属蛋白酶9及其特异性组织抑制因子的表达及临床意义

目的:探讨基质金属蛋白酶9(MMP-9)、金属蛋白酶特异性组织抑制因子1(TIMP-1)在子宫内膜异位症患者内膜标本中表达水平的变化及其临床意义。方法:收集子宫内膜异位症患者的异位内膜56例(观察组)和非内异症患者的子宫内膜标本18例(对照组),采用RT-PCR半定量方法检测内膜标本中MMP-9、TIMP-1 mRNA表达率及表达强度。结果:观察组和对照组所有内膜标本均有TIMP-1 mRNA表达;观察组和对照组标本中MMP-9 mRNA的表达率分别为58.9%和50%;观察组异位内膜中MMP-9 mRNA的表达强度高于对照组(P<0.05);TIMP-1 mRNA的表达强度低于对照组(P<0.05);对于MMP-9/TIMP-1比值,观察组明显高于对照组(P<0.01),早期(I-II期)明显高于III、IV期(P<0.01)。结论:子宫内膜异位症患者内膜中TIMP-1 mRNA的表达减弱,降低了对MMP-9的抑制作用;同时子宫内膜异位症患者内膜MMP-9 mRNA的表达增强,加强了其侵袭能力,易于发生种植转移,这些因素可能在子宫内膜异位症的发生发展过程中发挥重要作用。     

MMP-2和TIMP-2在大肠癌组织中的表达及其意义

目的探讨基质金属蛋白酶(MMP-2)及其抑制物(TIMP-2)的表达与大肠癌浸润,转移的关系。方法应用免疫组织化学S-P法对58例大肠癌组织中MMP-2和TIMP-2进行检测,并以大肠腺瘤组织标本20例作为对照。结果MMP-2的表达率在大肠癌为74.1%,高于大肠腺癌瘤的35%(P<0.05)。其表达与大肠癌的浸润深度,Dukes分期和淋巴结转移均密切相关(P<0.05)。TIMP-2的表达在大肠腺瘤及大肠癌组织中无显著性差异(P>0.05),但其表达与大肠癌组织学类型。分化程度,淋巴结有无转移密切相关,大肠癌MMP-2和TIMP-2蛋白表达存在显著负相关(r=-0.3684,P<0.05)。结论MMP-2和TIMP-2对大肠癌浸润、转移均有明显影响。MMP-2蛋白阳性表达促进大肠癌浸润和转移,MMP-2和TIMP-2的检测可作为临床判断大肠癌的恶性程度、转移及估计预后的重要参考指标。     

MMP-2和TIMP-2在大肠癌组织中的表达及其意义

目的 探讨基质金属蛋白酶(MMP-2)及其抑制物(TIMP-2)的表达与大肠癌浸润,转移的关系.方法 应用免疫组织化学S-P法对58例大肠癌组织中MMP-2和TIMP-2进行检测.并以大肠腺瘤组织标本20例作为对照.结果 MMP-2的表达率在大肠癌为74.1%,高于大肠腺癌瘤的35%(P<0.05).其表达与大肠癌的浸润深度,Dukes分期和淋巴结转移均密切相关(P<0.05).TIMP-2的表达在大肠腺瘤及大肠癌组织中无显著性差异(P>0.05),但其表达与大肠癌组织学类型.分化程度,淋巴结有无转移密切相关.大肠癌MMP-2和TIMP一2蛋白表达存在显著负相关(r=-0.3684,P<0.05).结论 MMP-2和TIMP-2对大肠癌浸润、转移均有明显影响.MMP-2蛋白阳性表达促进大肠癌浸润和转移,MMP-2和TIMP-2的检测可作为临床判断大肠癌的恶性程度、转移及估计预后的重要参考指标.     

The 372 T/C genetic polymorphism of TIMP-1 is associated with serum levels of TIMP-1 and survival in patients with severe sepsis

Introduction

Previous studies have found higher circulating levels of tissue inhibitor of matrix metalloproteinase (TIMP)-1 in nonsurviving septic patients than in surviving septic patients, and an association between the 372 T/C genetic polymorphism of TIMP-1 and the risk of developing certain diseases. However, the relationship between genetic polymorphisms of TIMP-1, circulating TIMP-1 levels and survival in patients with severe sepsis has not been examined, and this was the objective of the study.

Methods

This multicentre, prospective, observational study was carried out in six Spanish ICUs. We determined the 372 T/C genetic polymorphism of TIMP-1 (rs4898), serum levels of TIMP-1, matrix metalloproteinase (MMP)-9, MMP-10, TNFα, IL-10 and plasma plasminogen activator inhibitor-1 (PAI-1). Survival at 30 days from ICU admission was the endpoint assessed. The association between continuous variables was carried out using Spearman''s rank correlation coefficient or Spearman''s rho coefficient. Multivariate logistic regression analysis was applied to determine the association between the 372 T/C genetic polymorphism and survival 30 days from ICU admission.

Results

Of 275 patients with severe sepsis, 80 had genotype CC, 55 had genotype CT and 140 had genotype TT of the 372 T/C genetic polymorphism of TIMP-1. Patients with the T allele showed higher serum levels of TIMP-1 than patients without the T allele (P = 0.004). Multiple logistic regression analysis showed that the T allele was associated with higher mortality at 30 days (odds ratio = 2.08; 95% confidence interval = 1.06 to 4.09; P = 0.03). Survival analysis showed that patients with the T allele presented lower 30-day survival than patients without the T allele (χ² = 5.77; P = 0.016). We found an association between TIMP-1 levels and levels of MMP-9 (ρ = -0.19; P = 0.002), MMP-10 (ρ = 0.55; P <0.001), TNFα (ρ = 0.56; P <0.001), IL-10 (ρ = 0.48; P <0.001) and PAI-1 (ρ = 0.49; P <0.001).

Conclusion

The novel findings of our study are that septic patients with the T allele in the 372 T/C genetic polymorphism of TIMP-1 showed higher serum TIMP-1 levels and lower survival rate. The determination of the 372 T/C genetic polymorphism of TIMP-1 thus has prognostic implications and could help in the selection of patients who may benefit from modulation of the MMP/TIMP balance.