PDGF对人RPE细胞中MMP-2、MMP-9和TIMP-1表达的影响

背景研究表明血小板源性生长因子（PDGF）能调节多种细胞中基质金属蛋白酶／基质金属蛋白酶组织抑制剂（MMP／TIMP）的表达和平衡,但视网膜色素上皮（RPE）细胞中MMP／TIMP的表达与PDGF作用剂量和作用时间的关系尚不明确。目的观察PDGF对RPE细胞中MIP-2、MMP-9和TIMP-1表达的影响。方法体外培养人RPE细胞系ARPE-19,将达到70％～80％融合的细胞分为5个组。分别将0、0．1、1、10、50mg／LPDGF加入RPE细胞培养基作用36h,分别采用逆转录PCR（RT—RCR）法和Western blot法检测各组RPE细胞中MMP-2、MMP-9和TIMP-1 mRNA及其蛋白的表达。PDGF组采用10mg／LPDGF组PDGF分别刺激RPE细胞24、36和48h,对照组用不含PDGF的培养液培养,采用逆转录PCR（RT—RCR法和Western blot法分别检测RPE细胞中MMP-2、MMP-9和TIMP-1 mRNA及蛋白的表达。结果随着PDGF质量浓度的增加和刺激时间的延长,RPE细胞生长速度加快,细胞增生明显。PDGF刺激RPE细胞36h,随着PDGF质量浓度的增加,MMP-2 mRNA和MMP-9 mRNA在RPE细胞中表达相对值逐渐增加,各组间差异均有统计学意义（MMP-2 mRNA：F=79．304,P=0．000;MMP-9 mRNA：F=8．465,P=0．003）,其中1、10、50mg／LPDGF组RPE细胞中MMP-2 mRNA和MMP-9 mRNA表达相对值明显高于0mg／L PDGF组,差异均有统计学意义（P〈0．05）。RPE细胞中MMP-2和MMP-9蛋白表达相对值随着PDGF质量浓度的增加而逐渐增加,各组间差异均有统计学意义（MMP-2：F=26．550,P=0．000;MMP-9：F=80．993,P=0．000）,其中1、10、50mg／LPDGF组RPE细胞中MMP-2和MMP-9蛋白表达相对值明显高于0mg／L PDGF组,差异均有统计学意义（P〈0．05）。各质量浓度PDGF组RPE细胞中TIMP-1 mRNA和蛋白表达相对值差异均无统计学意义（F=0．143,P=0．962;F=1．955,P=0．178）。随着10mg／L PDGF刺激RPE细胞时间的延长,RPE细胞中MMP-2 mRNA和MMP-9 mRNA表达逐渐增加,差异均有统计学意义（MMP-2 mRNA：F时间=83．250,P=0．002;MMP-9 mRNA：F时间=6．785,P=0．019）;各时间点RPE细胞中MMP-2和MMP-9蛋白表达相对值明显增加,差异均有统计学意义（MMP-2：F时间=11．185,P=0．041;MMP-9：F时间=968．413,P=0．000）。PDGF作用不同时间点对照组与PDGF组间MMP-2、MMP-9 mRNA及蛋白在RPE细胞中的表达差异均有统计学意义（分组：均P=0．000,时间点：P〈0．05）,而各时间点2个组间RPE细胞中TIMP-1表达相对值的差异均无统计学意义（P〉0．05）。结论PDGF上调PRE细胞中MMP-2和MMP-9的表达,其作用呈剂量和时间依赖性,但对PRE细胞中TIMP-1的表达无明显影响。PDGF导致RPE细胞中MMP／TIMP的平衡失调,从而导致细胞外基质的破坏,促进RPE细胞的迁移。     

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.     

结膜瓣覆盖治疗角膜碱烧伤中MMP-9和TIMP-1的表达

目的：研究结膜瓣覆盖术治疗兔角膜碱烧伤中基质金属蛋白酶-9（MMP-9）及组织型基质金属蛋白酶抑制剂-1（TIMP-1）的表达机制。方法：将50只兔随机分成实验组及对照组,每组分别25只,建立兔角膜烧伤模型。实验组在角膜烧伤后当天行结膜瓣覆盖术。采用免疫组化法测定两组角膜碱烧伤后不同时间点MMP-9和TIMP-1的表达。结果：MMP-9在角膜碱烧伤后的3d开始升高,14d达到最高,之后逐渐下降;而TIMP-1在伤后即有表达,7d有所下降,于14d达到最低,21d达到峰值。实验组MMP-9的表达均明显低于对照组,且角膜碱烧伤后的3,14,21d和28d差异有统计学意义（P〈0.05）;而TIMP-1则明显高于对照组,且角膜碱烧伤后3,14d和21d差异有统计学意义（P〈0.05）。结论：角膜碱烧伤的病理损伤及修复过程中,MMP-9及TIMP-1的表达密切相关。结膜瓣覆盖治疗重度角膜碱烧伤的疗效确切。     

翼状胬肉中MMP-2,TIMP-2和TGF-β1的表达意义

目的:探讨MMP-2及其抑制剂TIMP-2和影响它们作用的TGF-β1在翼状胬肉中的表达及意义。方法:采用Maxvision免疫组化技术检测在20例翼状胬肉和10例正常球结膜中MMP-2、TIMP-2及TGF-β1的表达,并比较两组中表达的差异。结果:MMP-2和TGF-β1在翼状胬肉中的表达的阳性率显著高于正常球结膜中它们的阳性表达率(z=-4.618,-4.376,P<0.01),而TIMP-2在两组中的表达差异无显著性(z=-1.319,P>0.05)。经Spearman等级相关检验分析,在翼状胬肉中,MMP-2和TGF-β1的阳性表达率存在正相关(r=0·686,P<0.01);而MMP-2和TGF-β1与TIMP-2阳性表达率之间无相关(r=-0.410,-0.143,P>0.05)。结论:TGF-β1介导的MMP-2和TIMP-2表达失衡在翼状胬肉的发生、发展及侵犯角膜过程中发挥着重要的作用。     

Matrix metalloproteinases (MMP-2 and 9) and tissue inhibitors of matrix metalloproteinases (TIMP-1 and 2) during the course of experimental necrotizing herpetic keratitis

To determine the distribution and activities of metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) during the course of experimental herpes simplex virus (HSV) type-1 keratitis, BALB/c mice were corneally infected with 10(5) plaque-forming units (PFU) of HSV-1 (KOS strain) and then observed for the clinical signs of keratitis. Corneas were harvested at days 0, 2, 7 and 14 post-infection (p.i.). MMP-2, MMP-9, MMP-8, TIMP-1 and TIMP-2 were detected by immunohistochemistry and the Western blot technique. The enzymatic activities were analyzed by zymography. Epithelial HSV keratitis was present at day 2 after corneal infection and healed by day 5 p.i. While the expression and activity of MMP-2, MMP-8 and MMP-9 increased in the corneas at day 2 p.i., it was reduced at day 7 p.i. TIMP-1 and -2 were expressed in the corneas before and seven days after infection. Necrotizing stromal keratitis with corneal ulceration and dense polymorphonuclear leukocyte (PMN) infiltration was present at day 14 p.i. This correlated with increased expression of MMP-2, MMP-8 and MMP-9 in the corneas. MMP-8, MMP-9 and MMP-2 staining was particularly intense in the proximity of the ulcers and in areas of PMN infiltration. At day 14 p.i., MMP-2, -8 and -9 activities were upregulated, and TIMP-2 was expressed. These data suggest that MMPs produced by resident corneal cells and PMNs may possibly play a role in early epithelial keratitis and in the ulcerative process in the late phase after corneal HSV-1 infection. The ratio of MMPs to TIMPs may be important for the course of necrotizing HSV keratitis. TIMPs might participate in the repair process.     

Matrix metalloproteinases (MMP-8, MMP-9) and the tissue inhibitors of metalloproteinases (TIMP-1, TIMP-2) in patients with fungal keratitis

PURPOSE: To study the infiltrating cells and quantify the levels of matrix metalloproteinases (MMP-8, MMP-9) and tissue inhibitor of metalloproteinases (TIMP-1, TIMP-2) in the cornea, tear, and serum of patients with fungal keratitis. METHODS: Experimental study. Infected corneal tissue from 4 patients with fungal keratitis (group 1) scheduled to undergo therapeutic keratoplasty accounted for the histopathologic and cytospin smear analysis. Ten patients with fungal keratitis from group 2 served for the quantification of MMPs and TIMPs. Five patients with keratoconus undergoing penetrating keratoplasty and 5 cadaver corneas were chosen as controls for group 2. Corneal buttons obtained during keratoplasty, 15 to 20 microL of tears collected using the capillary flow method, and 3 mL of blood was obtained from patients with fungal keratitis and patients with keratoconus. Corneal button sections from group 1 were stained with hematoxylin and eosin and Grocott methenamine silver nitrate for the histopathologic studies and Giemsa staining for the cytospin smear analysis. Enzyme-linked immunosorbent assay was used for the quantification of total MMP-8, MMP-9, TIMP-1, and TIMP-2 in the corneal homogenates, tear, and serum samples of group 2. RESULTS: Corneal sections from group 1 revealed dense fungal filaments and a large proportion (91.4% +/- 38%) of polymorphonuclear leukocytes (PMNs). Significant elevation in the levels of MMP-8 and MMP-9 (P < 0.05) in the fungal keratitis corneas was observed in group 2 compared with the cadaver and keratoconus corneas. The ratio of MMP/TIMP was also higher in the fungal keratitis corneas. CONCLUSIONS: Infiltrating PMNs in the cornea of patients with fungal keratitis contributed to the increased activities of MMP-8 and MMP-9, thereby enhancing tissue destruction and derangement.     

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.     

Matrix metalloproteinase (MMP-2, -9) and tissue inhibitor (TIMP-1, -2) activity in tear samples of pediatric type 1 diabetic patients

Background

The presence of matrix metalloproteinase (MMP-2, -9) and tissue inhibitor (TIMP-1, -2) activity in tear samples of pediatric type 1 diabetes mellitus (DM) patients and potential correlations with clinical parameters (Schirmer testing, glycosylated hemoglobin-HB_A1C) were investigated.

Methods

Tear samples from the right eyes of 27 type 1 DM patients and 17 healthy control subjects were included in this study. The MMP gelatinolytic activity was determined by gelatin zymography analysis using sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE), while MMP and TIMP concentrations (in ng/ml) were quantified in tears of type 1 diabetic patients and healthy controls, with the use of enzyme-linked immunosorbent assay (ELISA).

Results

MMP-9, TIMP-1, -2 levels, MMP-9/TIMP-1, and MMP-9/TIMP-2 ratios in the patient group were significantly elevated. There was a significant correlation between TIMP-2 and HB_A1C values, as well as between MMP-2 and MMP-9.

Conclusions

Significant correlations between TIMP-2 and HB_A1C and between Schirmer test results and HB_A1C were revealed. Significant increase in tear MMP and TIMP levels in pediatric type 1 diabetic patients may be suggestive of disease progression and localized pathologic remodelling. Further studies are required in order to ascertain whether MMPs and TIMPs could be employed as indicators of early disease progression.     

Effects of Th2 cytokines on expression of collagen,MMP-1, and TIMP-1 in conjunctival fibroblasts

PURPOSE: To determine whether cytokines involved in chronic allergic conjunctival disorders may affect formation of giant papillae and tissue remodeling. METHODS: Conjunctival fibroblast cultures were challenged with different concentrations of human recombinant interleukin (IL)-4, IL-13, interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha. Procollagens I (PIP) and III (PIIIP), matrix metalloproteinase (MMP)-1 and -9, and tissue inhibitor of metalloproteinase (TIMP)-1 were measured in supernatants, and their respective mRNAs were evaluated by RT-PCR. RESULTS: IL-4 and -13 (10 ng/mL) significantly increased production and expression of PIP compared with nonstimulated cells, whereas IFN-gamma elicited the opposite effect, at both the protein and mRNA levels. Both IL-4 and -13 significantly decreased production of MMP-1 and increased that of TIMP-1, whereas TNF-alpha increased production of MMP-1 and -9. Expression of MMP-1 was reduced by IL-4 and increased by the other tested cytokines, whereas expression of TIMP-1 was increased by all tested cytokines. CONCLUSIONS: IL-4 and -13 increased production of collagen and modified the equilibrium between MMP-1 and its inhibitor, TIMP-1. These effects were partially opposed by IFN-gamma and TNF-alpha.     

外伤性PVR大鼠视网膜中MMP-9、TIMP-1的表达及意义

目的:为研究外伤性PVR和外伤后应用GM6001干预大鼠视网膜组织MMP-9及TIMP-1、在不同病程中的表达变化。方法:360只SD大鼠随机分为正常对照组、外伤性PVR组和外伤后应用GM6001组。正常对照组玻璃体腔内注射生理盐水;外伤性PVR组玻璃体腔内注射PRP血浆制成外伤性PVR大鼠动物模型;外伤后应用GM6001组在外伤后12h玻璃体腔内注射GM6001。应用免疫组化染色方法分别于1,3,7,14,21,28d对各组大鼠视网膜组织MMP-9及TIMP-1的表达检测。结果:免疫组化结果示MMP-9、TIMP-1蛋白均主要表达于视锥视杆层、视网膜内外网状层、神经纤维层。MMP-9在正常对照组、外伤后应用GM6001组的各个亚组微弱表达。MMP-9在外伤性PVR组1,3,7d显著表达,与正常对照组和外伤后应用GM6001组的差异有显著性(P(0.01),随着病程的延长,MMP-9的表达呈进行性减弱的趋势;TIMP-1在外伤性PVR组与外伤后应用GM6001组的各个亚组均有明显表达,与正常对照组的差异均有显著性(P(0.01)。MMP-9/TIMP-1比率在外伤性PVR组1,3,7d增高,与正常对照组和外伤后应用GM6001组的差异均有显著性(P(0.05)。结论:MMP-9、TIMP-1参与了PVR发生发展的病理过程,MMP-9/TIMP-1比率增高促进PVR发生发展的进程。人工合成基质金属蛋白酶抑制剂GM6001可促进MMP-9/TIMP-1动态平衡的重新建立,从而在外伤性PVR的防治中起重要作用。     

近视鸡眼巩膜纤维层MMP-2和TIMP-2的表达

目的　检测鸡眼巩膜纤维层MMP 2和TIMP 2的表达 ,进一步阐明近视鸡眼巩膜重塑的机制。方法　采用形觉剥夺的方法建立鸡眼近视模型。提取鸡眼巩膜纤维层的总RNA和蛋白质。利用RT PCR和Westernblot技术检测MMP 2和TIMP 2的mRNA和蛋白质表达水平的改变。结果　近视组鸡眼巩膜纤维层MMP 2mRNA和蛋白质表达水平明显升高 ,分别是正常组的 2 14和 2 11倍 ,差异显著 (P <0 0 1,P <0 0 1) ,而TIMP 2的表达水平较正常组分别下降5 5 0 5 %和 5 3 73 % ,差异亦显著 (P <0 0 1,P <0 0 5 )。结论　近视鸡眼巩膜纤维层变薄是组织主动重塑的结果 ,至少部分是由于MMP 2和TIMP 2表达改变所致。     

Corneal stimulation of MMP-1, -9 and uPA by platelet-activating factor is mediated by cyclooxygenase-2 metabolites

PURPOSE: This study was undertaken to evaluate the significance of cyclooxygenase-2 (COX-2) activity on urokinase plasminogen activator (uPA) and matrix metalloproteinases (MMPs)-1 and -9 induction in cornea following platelet-activating factor (PAF) treatment. METHODS: Corneal organ cultures were pre-treated with increasing concentrations of COX-2-specific inhibitors NS398 or nimesulide prior to PAF stimulation. To determine the effect of exogenous prostaglandins (PGs) on uPA, MMP-1 and MMP-9 levels, corneas were pre-treated with COX-2 inhibitors followed by the addition of 2.5 microM PGD2, PGE2 or PGF2alpha. The levels of uPA and MMP-9 were assayed by casein and gelatin zymography, respectively. MMP-1 levels were determined by Western Blot analysis. RESULTS: The increase in uPA, MMP-9 and MMP-1 levels detected in corneal organ cultures treated with 100 nM cPAF was blocked by 5 microM NS398 and 10 microM nimesulide, concentrations at which these inhibitors selectively inhibit COX-2 activity. Furthermore, pre-incubation with COX-2 inhibitors, followed by supplementation with PGD2, PGE2 or PGF 2alpha, increases uPA, MMP-9 and MMP-1 levels in corneas similar to and in some cases greater than that produced by cPAF treatment alone. CONCLUSIONS: During corneal injury and inflamation, PAF is an important factor in the activation of proteolytic cascades, which could lead to corneal epithelial defects and ultimately ulceration. One important goal in treating these defects is to modulate the activity of enzymes that destroy the extracellular matrix. Our results suggest that COX-2 induction following PAF stimulation and subsequent eicosanoid release may play a crucial role in the induction of uPA, MMP-1 and MMP-9 enzymes. Specific COX-2 inhibition could therefore block the actions of PAF when inflammation is sustained.     

透镜诱导豚鼠近视眼巩膜基质中基质金属蛋白酶2及其组织抑制剂和生长因子的变化

目的观察透镜诱导豚鼠近视眼巩膜基质中基质金属蛋白酶2（matrix metalloproteinases 2，MMP-2）、金属蛋白酶2组织抑制剂ftissue inhibitor of metaloproteinase 2，TIMP-2）和转化生长因子B2（transforming growth factorβ2，TGF—β2）和成纤维细胞生长因子（fibroblast growth factor，bFGF）的变化．探讨透镜诱导与形觉剥夺在近视眼发生机制上的异同点。方法 11只豚鼠于诱导前后分别验光、测量眼轴长度．用-6D的透镜诱导右眼作为诱导眼，左眼作为对照眼，诱导成功后，将豚鼠处死，摘除眼球，取后极部巩膜分别进行MMP-2和TIMP-2、TGF—β2和bFGF的免疫组织化学染色．观察它们在巩膜组织成纤维细胞胞浆中表达的阳性率和阳性细胞所占面积的百分比，并与对侧眼进行比较。结果11只豚鼠诱导眼成功诱导出（-3．31±1．04）D近视，诱导眼巩膜中MMP-2的阳性率为81．8％，对照眼为45．5％，差异有显著性；TIMP-2的阳性率36．4％，对照眼为72．7％；MMP-2染色阳性细胞面积百分比为0．22±0．15．对照眼为0．06±0．08（P〈0．05）；TIMP-2为0．05±0．08，对照眼为0．13±0．10。巩膜中TGF—β2阳性率为81．8％，对照眼为36．4％（P〈0．05），bFGF的阳性率为45．5％，明显低于对照眼（63．6％）。诱导眼TGF—β2染色阳性细胞面积百分比为0．21±0．14．对照眼为0．06±0．10，差异有显著性：bFGF为0．07±0．09，对照眼为0．15±0．14。结论透镜可以成功地诱导出豚鼠近视眼．透镜诱导近视眼巩膜基质中MMP-2活性增强，TIMP-2活性降低，TGF—β2表达增加，bFGF表达减少，MMP-2和TIMP-2、TGF—β2和bFGF之间存在平衡失调．它们的改变与形觉剥夺性近视的变化相似，表明透镜诱导近视眼与形觉剥夺性近视眼在巩膜基质的改变和生长因子的变化上存在相似的机制。     

NGF联合甲钴胺对青光眼视神经的保护作用及对MMP-2、TIMP-2的影响

目的：探讨神经生长因子(nerve growth factor,NGF)联合甲钴胺对青光眼术后视神经的保护作用及对基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)、金属蛋白酶组织抑制因子-2(tissue inhibitor of matrix metalloprotease-2,TIMP-2)的影响。

方法：选取行小梁切除术治疗的急性闭角型青光眼患者54例73眼。按照数字列表法随机分为对照组(30例35眼)与联合组(24例38眼)。对照组术后应用甲钴胺治疗,联合组术后应用NGF和甲钴胺联合治疗。观察两组患者术后用药前后视力、眼压、平均视敏度(mean light sensitivity,MS)、平均缺损(mean visual field defect,MD)、视神经纤维层(retinal nerve fibre layer,RNFL)厚度、视乳头杯/盘比、P100波潜伏期、P100波振幅、神经特异性烯醇化酶(neuron-specific enolase,NSE)、一氧化氮(nitric oxide, NO)、一氧化氮合酶(nitric oxide synthase,NOS)活性、MMP-2和 TIMP-2的情况。监测用药期间不良反应。

结果：与用药前比较,两组患者用药后视力、MS、P100波振幅明显提高,MD、P100波潜伏期明显下降,差异有统计学意义(P<0.05),但联合组的变化幅度明显大于对照组,差异有统计学意义(P<0.05)。两组患者用药前后眼压、RNFL、视乳头杯/盘比无明显变化,差异无统计学意义(P>0.05)。两组患者用药后NO、NOS、MMP-2水平升高,NSE、TIMP-2、TIMP-2/MMP-2下降,差异有统计学意义(P<0.05),但联合组的变化程度大于对照组,差异有统计学意义(P<0.05)。

结论：NGF联合甲钴胺能有效改善青光眼术后视功能,对MMP-2、TIMP-2有一定的调节作用。     

MMP-9、TIMP-1在大鼠视网膜缺血-再灌注损伤中的表达

目的 探讨高眼压视网膜缺血-再灌注(retina ischemia-reperfusion，RIR)损伤时大鼠视网膜基质金属蛋白酶9(matrix metalloproteinase 9，MMP-9)和组织金属蛋白酶抑制剂1(tissue inhibitor of metalloproteinase 1，TIMP-1)的动态变化及意义。方法 利用前房高压灌注法造成大鼠视网膜缺血1h，再恢复其供血，并于再灌注0h、3h、12h、24h、48h和72h后处死，采用免疫组化SP法和计算机图像分析系统检测视网膜MMP-9和TIMP-1的表达和分布。结果 RIR 0h视网膜MMP-9表达极弱，MMP-9阳性染色于RIR3h出现，12h迅速增加，24h达高峰，以后逐渐下降，72h接近正常。RIR各时段TIMP-1水平无明显改变。结论 MMP-9是参与RIR损伤的重要因素．     

TIMP-2基因转染FDM豚鼠后极部巩膜MMP-2蛋白早期动态表达

目的探讨外源性基质金属蛋白酶组织抑制剂-2(tissue inhibitor of matrix metalloproteinases,TIMP)对豚鼠形觉剥夺性近视(form deprivation myopia,FDM)眼后极部巩膜基质金属蛋白酶-2(matrix metalloproteinases-2,MMP-2)蛋白表达的影响。方法单眼形觉遮盖法制备FDM豚鼠右眼模型,45只豚鼠分为TIMP-2组、空质粒组和生理盐水组,每组15只,右眼脉络膜上腔内分别注射转染TIMP-2基因的脂质体、空质粒和生理盐水,左眼暴露为自身对照;另15只豚鼠持续遮盖右眼,不作任何处理,为对照组。各组分别于注药后的第2天、第7天和第14天处死豚鼠,取眼球后极部巩膜组织,用明胶酶谱法检测MMP-2蛋白的表达。结果转染第2天、第7天和第14天TIMP-2组豚鼠后极部巩膜MMP-2酶原及活性酶的相对表达量分别为0.9012±0.0056和0.3006±0.0051、0.8876±0.0060和0.2858±0.0065、0.8915±0.0068和0.2915±0.0076,表达均降低,与自身对照组、对照组组间比较,差异均有统计学意义(均为P<0.05),而空质粒组和生理盐水组与对照组相比,转染后第2天、第7天和第14天差异均无统计学意义(均为P>0.05)。TIMP-2组豚鼠后极部巩膜MMP-2酶原及活性酶表达水平从第2天开始降低,第7天最低,第14天时略有回升,第2天与第7天、第14天之间差别均有统计学意义(均为P<0.05),第7天与第14天比较差异无统计学意义(P>0.05)。结论外源性TIMP-2基因注入FDM豚鼠后,早期即可有效抑制后极部巩膜MMP-2蛋白的表达,减缓巩膜的重塑,但随时间的延长抑制作用逐渐减弱。     

Changes in scleral MMP-2, TIMP-2 and TGFbeta-2 mRNA expression after imposed myopic and hyperopic defocus in chickens

Induction of myopia leads to a decreased glycosaminoglycan synthesis and smaller collagen fibrillar diameters, increased levels of gelatinase-A (MMP-2) and decreased amounts of tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) in the fibrous sclera of both chicks and tree shrews. Another factor found to be involved in altered eye growth is the transforming growth factor beta-2 (TGFbeta-2). The aim of the current study was to measure MMP-2, TIMP-2 and TGFbeta-2 mRNA expression changes separately in the two scleral layers of chicks, following myopic and hyperopic defocus. Chicks were treated unilaterally with positive and negative lenses for different time periods. All contralateral eyes wore plano lenses and additional controls, treated binocularly with plano lenses, were included. Real-time PCR was used to measure MMP-2, TIMP-2 and TGFbeta-2 mRNA levels. Few changes in MMP-2 and TIMP-2 mRNA levels were measured following treatment with plus and minus lenses for up to 3 days. The mRNA levels of MMP-2 and TIMP-2 were either unchanged or co-regulated in both eyes, even though only the eye with the powered lens actually displayed changes in growth. In contrast, TGFbeta-2 mRNA was significantly up-regulated in the cartilaginous layer following treatment with plus lenses after 24 hr, compared to all other groups. These changes were confined to the eyes that also displayed reduced growth, suggesting a role of TGFbeta-2 in the final steps of visual eye growth regulation.     

凉血化瘀中药与曲安奈德对脉络膜新生血管动物模型中MMP-9及TIMP-2表达的影响

目的:评价凉血化瘀中药及曲安奈德对氪激光诱导BN大鼠脉络膜新生血管(choroidal neovascularization CNV)中基质金属蛋白酶MMP-9和基质金属蛋白酶抑制剂TIMP-2表达的影响。方法:659nm激光诱导BN大鼠CNV模型,随机分成空白对照组(每日生理盐水灌胃)、中药组(每日中药灌胃)和曲安奈德组(激光后1d玻璃体腔注射曲安奈德5μL,0.2mg)各16只。分别在光凝后7,14,21,28d随机选取4只大鼠处死,摘除眼球,行病理切片及免疫组织化学检测MMP-9及TIMP-2蛋白在CNV组织中的表达。结果:空白对照组在光凝后7dMMP-9达到高峰,此后逐渐下降;中药组在光凝后14d达到高峰,但峰值低于空白对照组,此后逐渐下降;曲安奈德组在光凝后7d达到高峰,峰值介于空白对照组和中药组之间,此后逐渐下降,在14～21d出现平台期。空白对照组在光凝后14dTIMP-2达到高峰,此后逐渐下降;中药组在光凝后14d达到高峰,峰值高于空白对照组,此后逐渐下降;曲安奈德组在光凝后14d达到高峰,峰值介于空白对照组和中药组之间,此后下降。结论:在下调MMP-9、上调TIMP-2的表达进而抑制CNV形成方面,凉血化瘀中药的作用强于曲安奈德。