MMP-2及TIMP-2在宫颈癌组织中的表达及其对侵袭和转移的影响

目的 通过检测基质金属蛋白酶-2(MMP-2)及基质金属蛋白酶组织抑制物-2(TIMP-2)在宫颈癌中的表达,探讨其对宫颈癌侵袭转移的影响.方法 应用免疫组化SP法检测53例宫颈癌和14例正常宫颈组织中MMP-2和TIMP-2的表达情况,并进行*9字2检验.结果 免疫组化结果 显示MMP-2和TIMP-2在宫颈癌组织中...     

基质金属蛋白酶-9及其组织抑制物-1在妊娠输卵管黏膜中的表达

目的:检测基质金属蛋白酶-9(MMP-9)及其组织抑制物-1(TIMP-1)在输卵管黏膜中的表达,探讨与输卵管妊娠的关系。方法:采用免疫组织化学显色技术和图像半定量分析法,检测MMP-9和TIMP-1在人妊娠输卵管黏膜、人正常输卵管黏膜及正常宫内早孕子宫蜕膜组织中的表达。结果:MMP-9和TIMP-1在正常宫内早孕组中表达最强,在输卵管妊娠组中的表达较强,在正常输卵管组中表达较弱。两两比较差异均有显著性。结论:MMP-9/TIMP-1参与了输卵管妊娠中胚胎着床过程,且与输卵管妊娠缺乏蜕膜化反应有关。     

子宫腺肌症与肿瘤转移相关基因之间关系的研究

目的：探讨肿瘤转移相关基因在子宫腺肌症发生中的作用。 方法： 采用免疫组织化学方法，对43例子宫腺肌症患者、22例对照组（正常子宫内膜）的nm23-H1、基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶-9（MMP-9）、膜型1-基质金属蛋白酶（MT1-MMP）和基质金属蛋白酶组织抑制因子-1（TIMP-1）的表达进行研究。 结果： 子宫腺肌症中，MMP-2、MMP-9和MT1-MMP的表达水平明显高于对照组（P<0.01），nm23-H1和TIMP-1的表达水平无显著差异（P>0.05）。 结论： MMP-2、MMP-9和MT1-MMP在子宫腺肌症的发病过程中可能起重要的作用。     

缺氧对肺动脉成纤维细胞MMP-2、TIMP-1表达的影响

目的：探讨缺氧对肺动脉成纤维细胞（Fpa）分泌基质金属蛋白酶（MMPs）、金属蛋白酶组织抑制剂（TIMPs）的影响。 方法： 采用酶谱法测定Fpa培养基中MMP-2的酶活性，免疫印迹法检测培养基中MMP-2、TIMP-1 的蛋白水平，免疫组化法测定细胞原位的蛋白表达， RT-PCR法检测mRNA表达量。 结果： 缺氧后Fpa分泌的MMP-2酶活性、细胞内外蛋白表达量、mRNA表达量均下降；而TIMP-1的表达则呈相反变化。 结论： 缺氧可使肺动脉成纤维细胞MMP-2/TIMP-1的表达失衡，可能参与缺氧性肺血管重建。     

大肠癌及其转移灶中MMP-2和TIMP-2的表达和意义

目的研究基质金属蛋白酶-2（MMP-2）及其抑制物组织基质金属蛋白酶抑制剂-2（TIMP-2）在大肠癌进展中的作用及其临床意义。方法通过免疫组化的方法检测104例大肠癌组织及其转移灶中MMP-2和TIMP-2的表达。结果MMP-2在转移灶中的阳性率（72．1％）显著高于原发灶中的阳性率（53．8％）（P〈0．05）,而TIMP-2在转移灶中的阳性率（35．6％）显著低于原发灶中的阳性率（66．3％）（P〈0．05）;MMP-2和TIMP-2在大肠癌原发及转移灶中表达均具有相关性,且呈负相关。结论MMP-2和TIMP-2的表达与大肠癌转移密切相关,而且转移灶癌细胞MMP-2的表达明显增强,TIMP-2的表达明显下降,可作为临床判断大肠癌恶性程度、转移及预后的重要参考指标。     

TLR4介导高迁移率族蛋白致系统性红斑狼疮肾损害的作用研究

目的 探讨高迁移率族蛋白1(HMGB1)致红斑性狼疮肾损害的作用机制与Toll样受体4(Toll-like receptor 4,TLR4)表达的相关性.方法 ELISA检测12例健康对照组、16例系统性红斑狼疮(systemic lupus eqrthematosus,SLE)无肾脏损害和18例狼疮性肾炎(lupus nephritis,LN)患者血清中HMGB1、基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶组织抑制剂-2(TIMP-2)的表达情况;流式细胞术检测外周血CD3/TLR4和CD14/TLR4表达情况;分离外周血单个核细胞(PBMC),RT-PCR检测HMGB1 mRNA的表达变化.结果 HMGB1 mRNA相对表达量及血清中HMGB1蛋白在LN组明显高于SLE组和健康对照组;流式细胞术显示CD14+的单核细胞表面HMGB1受体TLR4在LN组表达最高(P＜0.05),且与尿蛋白呈正相关(P＜0.01);LN患者血清中MMP-2和TIMP-2蛋白的浓度明显低于SLE和健康对照组,同时MMP-2/TIMP-2比值下降.HMGB1 mRNA及CD14+/TLR4+与MMP-2/TIMP-2比值均呈显著负相关;LN组患者血清中HMGB1蛋白水平与蛋白尿呈正相关,与MMP-2/TIMP-2比值呈显著负相关.结论 HMGB1是狼疮性肾炎发病中的重要细胞因子;HMGB1可能部分通过TLR4激活PBMC,降低MMP-2/TIMP-2的活性,从而引起蛋白尿.     

MMP-2,TIMP-1在肝细胞癌中的表达及临床意义

目的:探讨基质金属蛋白酶-2(MMP-2)及抑制剂-1(TIMP-1)在人肝癌及癌旁组织中的表达及其与肝癌侵袭转移的关系.方法:应用免疫组织化学法研究50例肝癌及癌旁组织中MMP-2和TIMP-1的表达.结果:MMP-2在肝癌和癌旁组织表达阳性率分别为76%和72%,明显高于正常肝组织的表达(P<0.01)a TIMP-1在肝癌组织和癌旁组织中的表达阳性率分别为58%和56%,高于正常肝组织中的表达(P<0.01).而MMP-2洋相表达率与肝细胞癌的转移、淋巴浸润、包膜形成和肿瘤分化有关(P<0.05).而TIMP-1的表达与癌组织的转移,淋巴结浸润和包膜形成有关(P<0.05).结论:MMP-2,TIMP-1在肝癌及癌旁组织中的表达明显增高,可能对于判断肝癌的浸润、转移有重要价值.     

家兔慢性冬眠心肌MMP-2、MMP-9和TIMP-2表达与胶原重构

目的观察家兔慢性冬眠心肌(CHM)心肌间质胶原纤维变化及基质金属蛋白酶-2,9(MMP-2、MMP-9)及其抑制剂(TIMP-2)的表达。方法27只家兔随机分为模型组(CHM,n=15)及假手术组(SHAM,n=12)。观察心肌间质胶原容积百分比(ICVF)、Ⅰ型和Ⅲ型胶原纤维比例、计算Ⅰ/Ⅲ型胶原纤维比例记分(CRS);免疫组化染色及Western blot观察MMP-2、MMP-9及TIMP-2表达。结果与SHAM组相比,CHM组ICVF显著增加(P<0.01);Ⅰ/Ⅲ胶原比例增高;CRS显著增加(P<0.01);MMP-2及MMP-9的表达明显增多(P<0.01),TIMP-2的表达明显减少(P<0.01)。结论在CHM过程中,MMPs/TIMPs比例失平衡,可能是CHM间质胶原重构的机制之一。     

子宫肌瘤患者中TIMP-3和IMMP-13的检测意义

目的比较UL（子宫肌瘤）组织和瘤旁子宫正常组织内的TIMP-3（基质金属蛋白酶抑制因子3）和MMP-13（基质金属蛋白酶一13）的表达差异及TIMP-3与MMP-13之间的相关性,以探讨UL的发生发展与两者间的关系。方法分析2012年4月至2014年4月在我院接受部分子宫切除或全切术治疗的UL患者的临床资料。用石蜡将手术切除的UL组织和瘤旁正常组织包埋做标本,免疫组织化学染色SP法检测TIMP-3和MMP-13的表达。比较UL组织和瘤旁组织的TIMP-3、MMP-13的阳性表达率,并分析TIMP-3与mmP-13表达间的相关性。结果本研究共纳入UL患者50例,其中包括UL组织和癌旁正常组织各50个。UL组织的MMP-13阳性表达率显著高于瘤旁组织（X//62=36．24,P〈0．01）;UL组织的TIMP-3阳性表达率显著高于瘤旁组织（X^2=9．632,P=0．022）;UL组织中mmP-13和TIMP-3的表达情况两者间有着显著的相关性（r=0.603,P=O．021）。结论UL的发生发展与TIMP-3和mmP-13的关系密切。     

N-乙酰基-丝氨酰-天冬氨酰-赖氨酰-脯氨酸对基质金属蛋白酶-1和基质金属蛋白酶组织抑制因子-1的调节在大鼠矽肺纤维化形成中的作用

目的:探讨N-乙酰基-丝氨酰-天冬氨酰-赖氨酰-脯氨酸(AcSDKP)对大鼠肺内基质金属蛋白酶-1(MMP-1)和基质金属蛋白酶组织抑制因子-1(TIMP-1)表达的调节在拮抗矽肺纤维化形成过程中的作用.方法:气管内灌注染尘法制作大鼠矽肺模型,将含有AcSDKP的微量药物释放泵埋入腹腔.实验动物随机分为矽肺模型对照4周组,矽肺模型对照8周组,矽肺模型4周组,矽肺模型8周组,抗纤维化治疗组及预防治疗组.H-E染色和免疫组织化学显色对矽肺纤维化病变和MMP-1和TIMP-1在肺组织内的表达进行形态学观察;免疫印迹法对肺内MMP-1和TIMP-1酶蛋白表达进行检测.结果:与模型对照组比较,矽肺大鼠肺内MMP-1和TIMP-1表达增强.与矽肺模型组比较,AcSDKP能够上调矽肺大鼠肺内MMP-1的表达,下调TIMP-1的表达,使MMP-1/TIMP-1的比值升高.结论:AcSDKP能够促进矽肺大鼠肺内MMP-1的表达,抑制TIMP-1的表达,从而加速了细胞外基质(包括胶原)的降解,这可能与AcSDKP抗矽肺纤维化的作用有关.     

The role of matrilysin (MMP-7) in leukaemia cell invasion

The matrix metalloproteinases (MMPs) are important in tumour cell invasion and metastasis in many common cancers. However, relatively few studies have investigated the role of MMPs and their inhibitors, the tissue inhibitors of metalloproteinases (TIMPs), in leukaemia cell invasion. This study examined two leukaemia cell lines, K562 and HL-60 and showed that the K562 cell line was four times more invasive than the HL-60 cell line. The expression of MMP-2, matrilysin (MMP-7), MMP-9, TIMP-1, TIMP-2 and TIMP-3 was analysed. Both cell lines produced similar amounts of MMP-2, MMP-9 and TIMP-2. The K562 cells expressed more TIMP-1 than the HL-60 cells and neither cell line expressed TIMP-3. Interestingly, only the K562 cells expressed matrilysin suggesting a potential role for matrilysin in leukaemia cell invasion. In vitro invasion assays performed in the presence of a matrilysin blocking antibody showed a 40% reduction in invasive ability. This data suggests that matrilysin plays an important role in leukaemia cell invasion. This revised version was published online in July 2006 with corrections to the Cover Date.     

基质金属蛋白酶及其组织抑制因子在左心室机械辅助减负荷模型中的表达

目的 研究基质金属蛋白酶(MMP)及其组织抑制因子(TIMP)在左心室机械辅助减负荷模型中的表达,探讨左心室减负荷后心肌逆向重构的分子机制.方法 结扎Lewis大鼠冠状动脉左前降支诱导心力衰竭,4周后将14只心力衰竭大鼠随机分为心力衰竭组(n=7)与移植组(n=7).将供体移植组心力衰竭大鼠的心脏及右肺移植到受体正常Lewis大鼠的腹部,通过供体的升主动脉与受体的降主动脉吻合.7只正常Lewis大鼠作为正常组.结扎左前降支4周后心脏超声测量3组大鼠心室直径和心肌梗死范围.移植2周后,称取各组大鼠心脏、左心室质量;显微镜观测左心室心肌细胞直径与心肌纤维化程度;采用实时荧光定量PCR检测MMP-1、MMP-9、TIMP-1的mRNA表达及计算MMP-1mRNA/TIMP-1 mRNA的比值.结果 结扎左前降支4周后,心力衰竭组及移植组舒张末直径(LVEDD)较正常组升高、左心室短轴缩短率(LVFS)较正常组下降,而此两组间LVEDD、LVFS及心肌梗死范围比较差异无统计学意义,两组的心力衰竭严重程度差异也无统计学意义.心力衰竭组心脏、左心室质量和左心室心肌细胞直径大于移植组与正常组;移植组心脏、左心室质量、左心室心肌细胞直径接近正常组.心肌纤维化的程度移植组＞心力衰竭组＞正常组[(7.90±2.32)％比(4.20±1.84)％比(1.54±0.31)％,均P＜0.05].心力衰竭组和移植组MMP-1、MMP-9mRNA表达均高于正常组(1.89±0.23、1.32±0.16比0.41±0.01,2.03±0.15、1.50±0.13比0.46+0.01,均P＜0.05),但心力衰竭组与移植组比较差异无统计学意义.心力衰竭组TIMP-1mRNA表达低于正常组与移植组(0.72±0.18比1.21±0.02、1.68±0.21,均P＜0.05);正常组与移植组比较差异无统计学意义.心力衰竭组MMP-1 mRNA/TIMP-1mRNA比值较正常组及移植组明显增高(2.03±0.15比0.30±0.01、0.81±0.11,均P＜0.05);正常组与移植组差异则无统计学意义.结论 左心室减负荷后,心肌逆向重塑的过程伴随着心肌细胞MMP及TIMP水平的改变.     

Role of metalloproteinases and tissue inhibitors of metalloproteinases during cardiopulmonary bypass in rats

Matrix metalloproteinases (MMPs) and the tissue inhibitors of matrix metalloproteinases (TIMPs) regulate matrix remodeling in the heart. Changes in synthesis and release of MMPs and TIMPs are observed after extracorporeal circulation (ECC). Thus, MMPs and TIMPs are supposed to be involved in ECC-mediated cardiac dysfunction. The aim was to examine the role of MMPs and TIMPs in ECC-mediated cardiac dysfunction. Extracorporeal circulation was instituted in rats for 60 min at a flow rate of 120 ml/kg/min. Three groups (n = 10) were studied: group CAO: 60 min ECC without aortic cross-clamping, group CAC: 60 min ECC including 30 min aortic cross-clamping (crystalloid Inzolen(?) cardioplegia), and group CAB: 60 min ECC including 30 min aortic cross-clamping (blood cardioplegia). Left ventricular (LV) function was measured with conductance catheter. Matrix metalloproteinase-activity was determined by zymography and TIMP activity was determined by reverse zymography. Gene expression of MMPs and TIMPs was determined by real-time polymerase chain reaction. Sixty minutes after weaning from bypass, there was a preserved LV function in the CAO and CAB group and an impaired LV function in the CAC group. We observed an increased myocardial activity and an increased myocardial messenger RNA expression of MMP-2, MMP-9, TIMP-1, and TIMP-4 in all ECC groups, when compared with sham animals. With regard to enzyme activity, there was an imbalance of MMP/TIMP ratio leading to an increased activity of MMP in the CAC group. In terms of gene expression, there was an imbalance of MMP-2/TIMP-4 ratio leading to an increased expression of MMP-2 in the CAC group. MMP-2 contributes to myocardial reperfusion injury in this in vivo model of ECC with cardioplegic arrest.     

螺内脂降低大鼠心肌非梗死区MMP/TIMP基因转录及表达

目的观察螺内脂对大鼠心肌非梗死区组织中MMP/TIMP活性和基因表达的影响,为心肌梗死的防治提供理论依据和新思路。方法建立大鼠药物干预的心肌梗死模型。用HE染色观察心肌梗死,用免疫组化及RT-PCR方法测定非梗死区心肌组织中MMP-2、MMP-9和TIMP-2的活性及表达。结果心肌梗死后48 h心肌梗死组非梗死区MMP-2、MMP-9和TIMP的表达明显升高,至第4周时仍维持在一个较高的水平(P<0.01)。螺内脂干预使MMP-2和MMP-9表达显著回降(P<0.01),而TIMP-2无明显下降。结论螺内酯可以降低大鼠非梗死区组织MMP-2、MMP-9/TIMP-2的活性及转录和表达。     

Regulation of matrix metalloproteinases and their inhibitors in the left ventricular myocardium of patients with aortic stenosis

Aortic stenosis (AS) results in myocyte and extracellular matrix remodeling in the human left ventricle (LV). The myocardial renin-angiotensin system is activated and collagens I and III and fibronectin accumulate. We determined the yet unknown regulation of enzymes that control collagen turnover, i.e., LV matrix metalloproteinases (MMP) and their tissue inhibitors (TIMPs) in human AS. We compared LV samples from AS patients undergoing elective aortic valve replacement (n=19) with nonused donor hearts with normal LV function (controls, n=12). MMP-2, MMP-9, MT1-MMP, and extracellular matrix metalloproteinase inducer (EMMPRIN), TIMP-1, TIMP-2, TIMP-3, and TIMP-4 mRNA were quantitated by real-time RCR. MMP-1, MMP-2, MMP-3, TIMP-3, TIMP-4, and EMMPRIN protein were measured by immunoblotting and MMP-9 and TIMP-1 protein by ELISA. Gelatinolytic MMP-2 and MMP-9 activity was measured by zymography. MMP-2 was increased in AS at mRNA, protein, and activity levels (131%, 193%, and 138% of controls). MMP-3 protein (308%) and EMMPRIN mRNA and protein were also upregulated (171% and 200%). In contrast, MMP-1 (37%) and MMP-9 mRNA, protein, and activity (26%, 21%, and 52%) were downregulated. MMP-9 activity was inversely correlated with LV size. TIMP-1 mRNA and protein were decreased (55% and 73%). In contrast, TIMP-2 mRNA (358%), TIMP-3 mRNA and protein (145% and 249%) were increased. TIMP-4 mRNA was not altered, but TIMP-4 protein was upregulated to 350%. Changes were similar in AS patients with normal and impaired LV ejection fraction. The dysregulation of myocardial MMPs and TIMPs in human AS starts at an early disease stage when LV function is still normal. In spite of upregulation of some MMPs the balance between MMP and TIMP is shifted towards MMP inhibition in human AS and may contribute to collagen accumulation.     

基质金属蛋白酶及其组织型抑制剂活性及表达失衡与高血压性左心室重塑的关系

目的： 探讨慢性压力负荷增高时，大鼠左心室（LV）基质金属蛋白酶（MMPs）/组织型基质金属蛋白酶抑制剂（TIMPs）失衡与LV重塑的关系。方法：40只6周龄雄性卒中易感性自发性高血压大鼠（SHR-SPs）作为研究对象，10只同周龄雄性Wistar-Kyoto（WKY）大鼠作为对照。6个月后，以Millar压力容积导管评价2组大鼠的在体LV血流动力学，并对2组大鼠的心脏进行组织病理学、明胶酶谱和免疫印迹法分析。结果：反映LV收缩与舒张功能的血流动力学参数在2组间有显著差异（P＜0.05）；SHR-SPs心脏胶原容积分数、血管周胶原面积/管腔面积、心肌横断面积、心室壁动脉中膜面积/管腔面积均增高（P＜0.05）；心肌MMP-2活性、蛋白含量及TIMP-1蛋白含量在SHR-SPs中明显增高（P＜0.05）。结论：慢性压力超负荷能够导致心脏细胞外基质代谢失调及MMPs/TIMPs系统失衡，继而产生心室腔扩张、LV收缩与舒张功能障碍。     

The effects of the initial treatment phase and of adjunctive low-dose doxycycline therapy on clinical parameters and MMP-8, MMP-9, and TIMP-1 levels in the saliva and peripheral blood of patients with chronic periodontitis

Introduction The treatment of periodontal disease can consist of bacterial plaque reduction, risk factor elimination, and metalloproteinase inhibitor medication. The level of matrix metalloproteinases (MMPs) are regulated by endogenous tissue inhibitors of metalloproteinases (TIMPs) as well as therapeutic low-dose doxycycline. The aim of the study was to evaluate the effect of the initial phase of periodontal treatment and the effect of doxycycline on clinical parameters and the MMP-8, MMP-9, and TIMP-1 concentrations in the saliva and peripheral blood of patients with chronic periodontitis. Materials and Methods The study group consisted of 33 patients with chronic periodontitis. Conventional periodontal treatment (scaling and root planing) was conducted on all the patients and doxycycline (20 mg orally) was administered twice daily for three months. Thirty-three controls received the conventional treatment only. Clinical scores (PI, BI, PD, CAL) were recorded before and three months after the treatment. MMP-8, MMP-9, and TIMP-1 concentrations in saliva and peripheral blood were measured by ELISA before and after the treatment of 20 patients from the study group and 13 of the controls. Results The application of doxycycline 20 mg resulted in significant improvement in clinical parameters compared with the conventional periodontal treatment. Doxycycline did not produce significant reductions in MMP-8 and MMP-9 levels in saliva observed after the conventional treatment. The study revealed increases in the TIMP-1 concentration and the MMP-8/TIMP-1 and MMP-9/TIMP-1 ratios in saliva and blood after treatment with doxycycline. Conclusions The study confirmed the modulating effect of doxycycline on the host response in chronic periodontitis.     

Tension Force Downregulates Matrix Metalloproteinase Expression and Upregulates the Expression of Their Inhibitors through MAPK Signaling Pathways in MC3T3-E1 cells

Objective: Matrix metalloproteinases (MMPs), produced by osteoblasts, catalyze the turnover of extracellular matrix (ECM) molecules in osteoid, and the regulation of MMP activity depends on interactions between MMPs and tissue inhibitors of metalloproteinases (TIMPs). We focused on the degradation process of ECM in osteoid that was exposed to mechanical strain, and conducted an in vitro study using MC3T3-E1 osteoblastic cells to examine the effects of tension force (TF) on the expression of MMPs and TIMPs, and activation of mitogen-activated protein kinase (MAPK) pathways.Design: Cells were incubated on flexible-bottomed culture plates and stimulated with or without cyclic TF for 24 hours. The expression of MMPs and TIMPs was examined at mRNA and protein levels by real-time RT-PCR and Western blotting, respectively. The phosphorylation of extracellular signal-regulated kinase (ERK) 1/2, p38 MAPK, and stress-activated protein kinases/c-jun N-terminal kinases (SAPK/JNK) were examined by Western blotting.Results: TF decreased the expression of MMP-1, -3, -13 and phosphorylated ERK1/2. In contrast, TF increased the expression of TIMP-2, -3 and phosphorylated SAPK/JNK. The expression of MMP-2, -14, TIMP-1, -4 and phosphorylated p38 MAPK was unaffected by TF. MMP-1, -3 and -13 expression decreased in cells treated with the ERK inhibitor PD98059 compared with untreated control cells. The JNK inhibitor SP600125 inhibited the TF-induced upregulation of TIMP-2 and -3.Conclusions: The results suggest that TF suppresses the degradation process that occurs during ECM turnover in osteoid via decreased production of MMP-1, -3 and -13, and increased production of TIMP-2 and -3 through the MAPK signaling pathways in osteoblasts.