MMP-2在眼组织中的表达与检测

基质金属蛋白酶-2（MMP-2）是基质金属蛋白酶（MMPs）家族中的一员，它的异常表达近年来被证明与多种疾病的发生发展有关，其在眼科疾病中起着重要作用。现笔者对MMP-2在眼组织中的表达、检测以及在眼病中的作用进行总结。     

MMP-2和MMP-9在视网膜母细胞瘤中的表达及意义

目的探讨视网膜母细胞瘤(retinoblastoma,Rb)中基质金属蛋白酶-2(matrixmetalloproteinase-2,MMP-2)和基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)的表达及其与临床病程的关系。方法应用免疫组织化学SP法检测41例Rb组织中MMP-2和MMP-9的表达;用HMIAS-2000型全自动医学彩色图像分析系统测量其平均光密度值。统计学方法比较不同临床和病理阶段Rb表达MMP-2和MMP-9的差异。结果在41例Rb组织切片中均有MMP-2和MMP-9的表达。有视神经浸润的肿瘤MMP-2和MMP-9表达水平明显高于无视神经浸润的肿瘤(P<0.01),处于眼外期的肿瘤MMP-2和MMP-9的表达水平明显高于眼内期和青光眼期的肿瘤(P<0.01)。结论Rb细胞内有MMP-2和MMP-9的表达。MMP-2和MMP-9的表达水平与视神经浸润和临床分期存在密切的关系,提示其表达水平与该肿瘤的浸润与发展有关。     

MMP-2、MMP-9的表达与脉络膜黑色素瘤微血管密度及肿瘤转移的关系

目的 探讨恶性脉络膜黑色素瘤细胞中基质金属蛋白酶-2（matrix metalloproteinase-2,MMP-2） 和基质金属蛋白酶-9（matrix metalloproteinase-9,MMP-9）的表达特征及其与血管生成的关系及预后意义.方法 采用免疫组织化学SP法检测56例（56眼）脉络膜黑色素瘤细胞中MMP-2、MMP-9的表达特征,以CD34标记检测微血管密度（microvessel density,MVD）并分析其与肿瘤血管生成及肿瘤转移的关系.结果 56例患者中MMP-2和MMP-9的表达阳性率较高,分别为91.07%和96.42%,两者的表达强度呈正相关关系（r=0.45,P=0.001）;有转移病例出现MMP-2、 MMP-9高表达的比例明显高于无转移病例（P=0.004,P=0.006）;MMP-2 、MMP-9高表达者的MVD明显大于其低表达者（P=0.025,P=0.021）.结论 MMP-2和MMP-9的过度表达可能与脉络膜黑色素瘤血管生成及肿瘤转移有关.     

基质金属蛋白酶与眼科疾病的研究进展

基质金属蛋白酶(matrix metalloproteinases,MMPs)属于Zn2+和Ca2+依赖性内肽酶家族,是参与降解细胞外基质(extracellular matrix,ECM)的最重要的蛋白酶。因其参与了多种眼科疾病的病理过程,故对其活化与抑制,以及如何高表达进行研究,可为预防和控制各类眼科疾病的发生发展提供崭新的研究方向,现就其最近的科研进展做一综述。     

基质金属蛋白酶及其抑制剂在眼表疾病发病机制研究中的进展

基质金属蛋白酶(matrix metalloproteinases,MMPs)是一组降解细胞外基质的内源性蛋白酶系,其与基质金属蛋白酶组织抑制剂(tissue inhibitor of metalloproteinases,TIMPs)组成MMPs/TIMPs系统,降解和重塑细胞外基质.MMPs/TIMPs系统表达水平的失衡与眼病的发生发展密切关联,尤其是在各类眼表疾病中.目前认为结膜成纤维细胞中MMP-1、MMP-3及MMP-9过度表达是引起MMPs与TIMPs之间失去平衡的关键因素.MMPs与TIMPs之间失去平衡,使胶原纤维融解,弹力纤维变性减少,导致球结膜基质和Tenon囊的过度降解,引起眼表泪液异常的病理循环.眼表泪液的异常破坏了眼表环境的稳定性,参与多个眼表疾病如干眼、结膜松弛症、翼状胬肉、角膜炎等的病理变化.     

基质金属蛋白酶及其组织抑制因子与眼病的相关性研究进展

基质金属蛋白酶(MMPs)是一组锌离子依赖的结构和功能同源的内肽酶家族,能降解细胞外基质(ECM)的多种成分.基质金属蛋白酶组织抑制剂(TIMP)是基质金属蛋白酶的特异性组织抑制物,是参与基质金属蛋白酶的活性调节的重要组织因子.它们共同参与多种重要的病理生理过程.目前国内外的许多研究发现MMPs和TIMP与眼科疾病的发生、发展和转归密切相关.人们对人工合成MMPs抑制剂也有很大的兴趣.     

假鳞皮样脱皮综合征患者房水中基质金属蛋白酶-1组织抑制因子和结缔组织生长因子水平升高

目的最近发现假鳞皮样脱皮综合征（pseudoexfoliation syndrome，PXF）患者房水中转化生长因子-β1（transforming growth factorβ1，TGFβ1）表达增高，因为抗TGFβ1可潜在的干扰前房相关免疫偏离的维持，针对抗TGFβ1治疗的关注日益增高，作者探究了前房中基质金属蛋白酶-1组织抑制因子（tissue inhibitor of matrix metalloproteinase-1，TIMP-1）、基质金属蛋白酶-9（matrix metalloproteinase-9，MMP-9）和结缔组织生长因子（connective tissue growth factor，CTGF）的水平，     

基质金属蛋白酶及其在眼科的研究进展

基质金属蛋白酶(MMPs)是一组金属离子依赖的结构和功能同源的内肽酶家族,能降解细胞外基质(ECM)的多种蛋白成分。它在眼科疾病中的作用已引起关注,本文就其结构、组成、功能、调控及其在眼科疾病中的作用做一综述。     

猪眼小梁细胞的鉴定及压力对猪眼小梁细胞MMP-2、MMP-3及TIMP-1表达的影响

目的 探讨压力对体外培养的猪眼小梁细胞基质金属蛋白酶-2(matrix metalloproteinases,MMP-2)、MMP-3以及基质金属蛋白酶组织抑制剂-1(tissue inhibitors of MMPs,TIMP-1)表达的影响.方法 培养猪眼小梁细胞并进行鉴定.对传第3代的小梁细胞施加20 mmHg(1 kPa=7.5 mmHg)、40 mmHg、60 mmHg及80 mmHg压力作为实验组,0 mmHg设为对照组.培养48 h后对MMP-2、MMP-3和TIMP-1进行免疫组织化学SP法染色及电镜扫描,并对染色结果进行统计学分析.结果 根据培养细胞的生长特性、形态特征及细胞免疫组织化学染色结果等特点,确定培养的细胞为猪眼小梁细胞.猪眼小梁细胞正常可以少量表达MMP-2、MMP-3和TIMP-1.20 mmHg、40 mmHg、60 mmHg时猪眼小梁细胞MMP-2的SP染色细胞阳性率分别为(34.30±10.89)%、(47.48±4.96)%、(58.40±8.67)%,TIMP-1的SP染色细胞阳性率分别为(20.40±7.63)%、(37.66±11.64)%、(49.58±8.10)%,MMP-3的SP染色细胞阳性率分别为(12.30±4.35)%、(11.98±2.26)%、(15.24±5.63)%.因此压力改变在一定程度上可以促进猪眼小梁细胞表达MMP-2、TIMP-1,而对猪眼小梁细胞MMP-3的表达无明显影响.结论 一定范围内压力的变化可以改变MMPs/TIMPs之间的平衡状态,进而影响小梁细胞外基质细胞外基质的代谢以及房水外流阻力,因此MMPs在青光眼的发病机制中发挥重要作用.     

基质金属蛋白酶及其在眼科的研究进展

基质金属蛋白酶(MMPs)是一组金属离子依赖的结构和功能同源的内肽酶家族，能降解细胞外基质(ECM)的多种蛋白成分。它在眼科疾病中的作用已引起关注，本就其结构、组成、功能、调控及其在眼科疾病中的作用做一综述。     

基质金属蛋白酶-9在人正常角膜及真菌性角膜溃疡中的基因表达

目的:研究基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)在人正常角膜及真菌性角膜溃疡中的基因表达,探讨MMP-9在真菌性角膜溃疡形成中的作用。方法:搜集24份真菌性角膜溃疡标本(溃疡组)及12份人正常角膜组织标本(对照组),其中12份真菌性角膜溃疡组织和6份正常的角膜组织分别抽提组织总的RNA,以Primer5.0软件设计MMP-9的特异引物,并对标本RNA进行RT-PCR扩增,以MMP-9目的电泳条带与β-actin内参照条带灰度值比值代表相对表达量。分12份角膜溃疡组织和6份正常的角膜组织进行免疫组化检测MMP-9表达,通过免疫组化染色的灰度值进行统计学分析。结果:MMP-9在真菌性角膜溃疡中的表达明显高于正常角膜,且主要位于角膜上皮层、基底膜以及基质层。结论:真菌性角膜溃疡组织中MMP-9的基因表达明显上调。MMP-9在真菌性角膜溃疡的发生、发展及角膜穿孔中起着重要作用。加强MMP-9在真菌性角膜溃疡中的作用机制的研究,有望对真菌性角膜溃疡的治疗提供新的途径。     

MMP-2和MMP-14在视网膜母细胞瘤中的表达及意义

目的:探讨视网膜母细胞瘤(retinoblastoma,Rb)中基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)和MMP-14的表达及其与临床病程的关系。方法:应用免疫组织化学SP法检测50例Rb组织中MMP-2和MMP-14的表达;用HMIAS-2000型全自动医学彩色图像分析系统测量其平均光密度值。统计学方法比较不同临床和病理阶段Rb表达MMP-2和MMP-14的差异。结果:患者50例Rb组织切片中MMP-2阳性表达38例,占76%;MMP-14的阳性表达41例,占82%;有视神经浸润的肿瘤MMP-2和MMP-14表达水平明显高于无视神经浸润的肿瘤(P<0.01),处于眼外期的肿瘤MMP-2和MMP-14的表达水平明显高于眼内期和青光眼期的肿瘤(P<0.01);Rb中MMP-2和MMP-14的阳性表达水平呈密切相关(P<0.01)。结论:MMP-2和MMP-14的表达水平与视神经浸润和临床分期存在密切的关系,提示其表达水平与该肿瘤的浸润与发展有关。     

Expression of MMP-2 and MMP-9 in retinoblastoma and their significance

AIM: To investigate the expression of matrix metallopr- oteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in retinoblastoma (Rb), and their relationships with tumor development stage. METHODS: Immunohistochemical technique was used to detect the expression of MMP-2 and MMP-9 in 41 cases of paraffin embedded Rb samples. Quantitative analysis of the expression of MMP-2 and MMP-9 was assessed by HMIAS-2000 Color Pathologic Analysis System. The differences of the expression of MMP-2 and MMP-9 in each clinical and pathological stage were analyzed statistically. RESULTS: In all the 41 Rb specimens, MMP-2 and MMP-9 expression was found in tumor cells. The expression of MMP-2 and MMP-9 was significantly higher in tumors with optic nerve invasion than in tumors without optic nerve invasion (P <0.05); the expression of MMP-2 and MMP-9 was significantly higher in tumors of extra-ocular stage than in tumors of glaucomatous stage or intra-ocular stage(P<0.05). CONCLUSION: MMP-2 and MMP-9 exist in retinoblastoma cells. The level of MMP-2 and MMP-9 is related to optic nerve invasion and clinical stage of Rb, which suggests the expression of MMP-2 and MMP-9 could be connected to the invasion and development of tumor cells. Further research is needed for deeper understanding of the biological behavior and better evaluation of the prognosis of Rb.     

Plasma levels of matrix metalloproteinase-2 and -9 (MMP-2 and MMP-9) in age-related macular degeneration

BACKGROUND: Several studies indicate that age-related macular degeneration (AMD) and atherosclerosis may share common pathogenetic pathways. The aim of this study was to determine the role of systemic matrix metalloproteinases (MMPs) in AMD, given that MMPs are implicated in the pathogenesis of atherosclerosis. METHODS: This study determined the plasma matrix metalloproteinases (MMP-2 and MMP-9) levels in three groups of subjects: group 1 included subjects with age-related maculopathy (ARM), group 2 included subjects with choroidal neovascularization (CNV) owing to AMD and group 3 consisted of age-matched controls. RESULTS: The mean plasma levels of MMP-2 were not significantly different in the three groups. In contrast, the mean plasma MMP-9 levels were significantly higher in ARM and CNV groups compared to that of the control group. However, there was no significant difference in MMP-9 levels between ARM and CNV groups. CONCLUSION: This is the first study that reveals a link between raised plasma MMP-9 levels with AMD. Further studies are required to identify the factors that contribute to this association.     

Plasma levels of matrix metalloproteinase-2 and -9 (MMP-2 and MMP-9) in age-related macular degeneration

BACKGROUND: Several studies indicate that age-related macular degeneration (AMD) and atherosclerosis may share common pathogenetic pathways. The aim of this study was to determine the role of systemic matrix metalloproteinases (MMPs) in AMD, given that MMPs are implicated in the pathogenesis of atherosclerosis. METHODS: This study determined the plasma matrix metalloproteinases (MMP-2 and MMP-9) levels in three groups of subjects: group 1 included subjects with age-related maculopathy (ARM), group 2 included subjects with choroidal neovascularization (CNV) owing to AMD and group 3 consisted of age-matched controls. RESULTS: The mean plasma levels of MMP-2 were not significantly different in the three groups. In contrast, the mean plasma MMP-9 levels were significantly higher in ARM and CNV groups compared to that of the control group. However, there was no significant difference in MMP-9 levels between ARM and CNV groups. CONCLUSION: This is the first study that reveals a link between raised plasma MMP-9 levels with AMD. Further studies are required to identify the factors that contribute to this association.     

Positive feedback regulation between MMP-9 and VEGF in human RPE cells

PURPOSE: The proteolytic activity of matrix metalloproteinases (MMPs) is involved in pathologic angiogenesis in the eye. However, it is unknown whether MMPs may stimulate the production of the major angiogenic factor, vascular endothelial growth factor (VEGF). The authors investigated whether MMP-2 and MMP-9 alter the expression of VEGF by retinal pigment epithelial (RPE) cells. They also sought to determine the effects of MMPs on cellular proliferation and migration and the effect of triamcinolone acetonide on MMP-9-evoked cellular responses. METHODS: Human RPE cell cultures were stimulated with MMP-2 or MMP-9. The gene expression and secretion of MMP-9 and VEGF were determined by real-time RT-PCR and ELISA, respectively. Cellular proliferation was investigated with a bromodeoxyuridine immunoassay, and chemotaxis was examined with a Boyden chamber assay. RESULTS: Under control conditions, RPE cells in vitro expressed a significantly higher amount of mRNA for MMP-2 than for MMP-9. Chemical hypoxia caused upregulation of the gene expression of both MMPs, whereas VEGF increased the gene expression and secretion of MMP-9. The hypoxic expression of MMP-9 was mediated by autocrine VEGF signaling. Exogenous MMP-9 increased the gene expression and secretion of VEGF, whereas MMP-2 reduced the secretion of VEGF. MMP-2 and MMP-9 did not alter the proliferation but stimulated the migration of RPE cells. Triamcinolone fully inhibited the stimulatory effect of MMP-9 on the expression of VEGF and the VEGF-evoked increase in the expression of MMP-9. However, triamcinolone had no effect on the motogenic effect of MMP-9. CONCLUSIONS: There is a positive feedback regulation between MMP-9 and VEGF in RPE cells. The hypoxic expression of MMP-9 may stimulate the production and secretion of VEGF under pathologic conditions. Triamcinolone inhibits the positive feedback regulation between MMP-9 and VEGF under hypoxic conditions through inhibition of the gene expression of MMP-9 and the secretion of VEGF.     

rd1 mouse retina shows imbalance in cellular distribution and levels of TIMP-1/MMP-9, TIMP-2/MMP-2 and sulfated glycosaminoglycans

BACKGROUND: The rd1 mouse retina displays fast degeneration of photoreceptors resulting in a depletion of almost all rod photoreceptors by postnatal day 21 (PN21). To evaluate the role of proteinases in the pathophysiology of this animal model of retinitis pigmentosa, C3H rd1 and congenic wild-type (wt) mice retinas were analyzed. MATERIAL AND METHODS: The cellular localization and levels of proteins, matrix metalloproteinases (MMPs), their endogenous inhibitors (TIMPs), total sulfated glycosaminoglycans (sGAG) and nature of saccharides in rd1 and wt retinal extracts were compared. RESULTS: MMP-2/TIMP-2 and MMP-9/TIMP-1 were predominantly localized in the interphotoreceptor matrix (IPM) of both genotypes, but MMP-2/TIMP-2 also appeared in the Muller cell fibers of rd1 retina. In rd1 retinal extracts the levels of total proteins were lower and those of active MMP-9, MMP-2, TIMP-1 and total sGAG were higher than those of wt extracts. Despite an increase in TIMP-1, active MMP-9/MMP-2 were disproportionately elevated in rd1 compared to wt retina. With increasing age, MMPs in wt retinas were decreased but were increased in rd1. The sialylation of proteoglycans in PN2 and PN7 rd1 retinas was lower, and galactosylation was higher than that in wt retinas. CONCLUSIONS: MMP-9/MMP-2 and TIMP-1/TIMP-2 are associated with IPM, possibly after secretion by retinal pigmented epithelial cells. In degenerating rd1 retina, MMP-2/TIMP-2 are associated with the Muller cell fibers, which apparently play a central role in modifying the balance between MMPs and TIMPs. Elevated sGAG and proteolysis due to an imbalance in the levels of TIMPs and active MMP-9/MMP-2 in rd1 retina possibly contribute to retinal degeneration in the rd1 mouse.     

热烧伤后的人角膜组织中基质金属蛋白酶-2、-9表达的研究

目的:研究热烧伤后人角膜组织中的基质金属蛋白酶-2、-9(MMP-2、MMP-9)的表达与分布,探讨其含量的变化与角膜病变的相关性。方法:收集铁水烫伤后行角膜移植患者病变角膜19例,正常角膜组织10例。应用免疫组化染色方法检测MMP-2、MMP-9在角膜各层次中的分布与表达,应用图像分析系统进行半定量分析。结果:MMP-2、MMP-9在正常角膜组织中均未见阳性表达,平均灰度值分别为117.8188±3.7967,119.1902±0.9679。热烧伤后角膜组织中,MMP-2的阳性表达主要分布于角膜基质层,平均灰度值为94.4197±5.8327;MMP-9的阳性表达主要分布于角膜上皮层和基底膜附近,平均灰度值为115.0188±0.7149。将正常角膜组织中和病变角膜组织中两种酶的平均灰度值分别进行统计学分析,P值均<0.05。随着病程的发展,在病变角膜组织中MMP-2的阳性表达先呈现逐渐增高趋势14d后逐渐下降,角膜基质层溶解达高峰时阳性表达的量最高;MMP-9在患病初期角膜基底膜病变明显时表达较高,后期逐渐下降。结论:MMP-2、MMP-9对热烧伤后的人角膜组织病变的发展起重要的作用。MMP-2主要参与角膜基质层的损害,MMP-9主要参与角膜上皮层和基底膜的损害。     

Estrogen up-regulation of metalloproteinase-2 and -9 expression in rabbit lacrimal glands

Increased levels of the matrix metalloproteinases (MMPs)-2 and -9 have been found in tear fluids of patients with dry eye disease, suggesting that these MMPs may be implicated in the pathogenesis of this disease. One of the main causes of dry eye disease is lacrimal gland insufficiency. However, the contribution of the lacrimal gland (LG) to the expression and production of MMP-2 and MMP-9 in tears is not known. Since dry eye disease occurs more frequently in women, sex hormones, especially estrogens, have also been implicated in the pathogenesis of this disease. Estrogens have been shown to regulate the synthesis levels of MMP-2 and MMP-9 in several tissues, Thus, the purpose of these studies was to determine if: (1) rabbit lacrimal glands secrete MMP-2 and MMP-9; (2) MMP-2 and MMP-9 are produced by lacrimal epithelial cells and/or lacrimal lymphocytes; and (3) the expression, activity and level of these enzymes are regulated by sex hormones. Lacrimal epithelial cells (LEC) and lacrimal lymphocytes (LL) from sexually mature New Zealand White female rabbits were isolated, purified and cultured with and without 10(-6)M dihydrotestosterone (DHT) or 10(-6), 10(-8), 10(-9) and 10(-10)M 17beta-estradiol (E2). The culture supernatants were analyzed by zymography and western blotting (WB) using polyclonal anti-human MMP-2 and MMP-9 antibodies. LGs were also collected from rabbits 7 days after being sham-operated, ovariectomized (OVX), OVX treated with 4 mg/kg DHT, and OVX treated with 0.5 mg/kg of E2. LGs were collected and processed for RNA extraction as well as protein determination using WB and immunocytochemistry. The pro-forms of MMP-2 and MMP-9 were detected in primary LEC and LL culture medium by zymography and WB. Pro-MMP-2 and pro-MMP-9 were also detected at the gene and protein levels in the lacrimal glands of all four treatment groups, with the highest levels and gene expression found in the estrogen-treated group. These results suggest that both pro-MMP-2 and pro-MMP-9 are secreted by the lacrimal gland and appear to be up-regulated by estrogen. The role of the lacrimal MMPs in the pathogenesis of dry eye disease needs to be further investigated.