Myocardial fibrosis in transforming growth factor-beta(1) (TGF-beta(1)) transgenic mice is associated with inhibition of interstitial collagenase

BACKGROUND: TGF-beta(1) mediates effects on fibroblast proliferation and collagen synthesis in the myocardium. The extracellular matrix remodeling depends on the fibrillar collagen degrading matrix metalloproteinases (MMPs) and their inhibitors (TIMPs). The in vivo effects of TGF-beta(1) on the MMP/TIMP system in TGF-beta(1) overexpressing transgenic mice were studied. METHODS: Male Alb/TGF-beta(1)(cys(223,225)ser) transgenic mice (TG) and nontransgenic controls (C; 8 weeks) were examined. Protein expression of collagen type I, -III, interstitial collagenase (Int Coll), MMP-2, -9, TIMP-1, -2, -4 and TGF-beta(1) as well as enzyme activity (MMP-2, -9) were measured (Western blots, zymographic assays). mRNA expression of the interstitial collagenase and MMP-9 was studied with the Light-Cycler based real-time PCR. RESULTS: Overexpression of TGF-beta(1) resulted in a 10-fold increase in plasma and a seven-fold increase in myocardial TGF-beta(1) concentrations. Relative heart weights increased (mg g(-1): 7.8 +/- 0.4 vs. 4.8 +/- 0.6, n = 6; P < 0.01) in TG compared to C. Collagen type I and III increased in TG (1.9-fold and 1.7-fold) compared to controls. Interstitial collagenase protein activity (- 91%) and mRNA expression (-75%) in TG were reduced (P < 0.05-P < 0.001). Gelatinase (MMP-2, MMP-9) expression and activity were not significantly alterated. MMP-inhibitors were increased 2.5-fold (TIMP-1, -4) and 6-fold (TIMP-2) in TG. CONCLUSIONS: TGF-beta(1) produces myocardial fibrosis in vivo. This effect is not only produced by a stimulation of matrix protein formation: a complex regulation of MMP and TIMP interaction, namely decrease of expression and activity of interstitial collagenase and an enhanced inhibition by increased levels of TIMPs, are involved. These mechanisms are optional targets for therapeutic interventions in myocardial diseases.     

基质金属蛋白酶及其组织抑制剂与滋养细胞侵袭性生长关系的研究

目的了解基质金属蛋白酶（MMPs）和组织抑制剂（TIMPs）在滋养细胞中的定位及其在不同滋养细胞病变中的表达和相互调节作用.方法采用免疫组化方法检测MMP-2、MMP-9和TMP-1、TIMP-2在绒毛膜癌、侵袭性水泡状胎块、水泡状胎块、胎盘植入和超常反应胎盘部位等病变以及胎盘着床部位中的表达.结果MMP-2、9和TIMP-1、2主要在中间滋养细胞和合体滋养细胞表达.滋养细胞在胎盘着床部位仅表达MMP-2;超常反应胎盘部位和胎盘植入不但表达MMP-2,而且多数病例MMP-9（＋）,但阳性强度弱,TIMP-1和TIMP-2（-）;水泡状胎块较强表达MMP-2,伴持续性滋养细胞疾病MMP-9和TIMP-1、2（＋）;侵袭性水泡状胎块和绒毛膜癌MMP-2和MMP-9表达明显增强,TIMP-1和TIMP-2多数（＋）.结论在病理性妊娠中MMP-9活性增强可导致滋养细胞发生过度浸润.MMP-9的过度表达和TIMP-1、2的轻微增加,可能共同增强了肿瘤细胞的侵袭能力.     

地塞米松对高氧肺损伤大鼠肺组织基质金属蛋白酶及其组织抑制剂表达的影响

目的 观察地塞米松对高氧暴露大鼠肺组织中基质金属蛋白酶(MMPs)及其组织抑制剂(TIMPs)表达的影响,探讨地塞米松治疗高氧肺损伤的作用机制。方法 2周龄Wistar大鼠32只,随机分为空气组和高氧组(各16只)。高氧暴露7 d后,取两组大鼠各8只检测支气管肺泡灌洗液(BALF)蛋白含量和肺湿/干重比(W/D),并观察肺组织病理学改变。余16只大鼠行肺组织培养,空气组8只作为空气对照组,高氧组8只各设高氧对照组,高氧 地塞米松1×10-8、1×10-6和1×10-4mol/L组,培养24 h后用逆转录-聚合酶链反应(RT-PCR)检测肺组织中MMP-2、MMP-9、TIMP-1、TIMP-2 mRNA的表达。结果①与空气组相比,高氧组肺组织出现水肿、出血、炎性细胞浸润;BALF中蛋白含量、W/D明显增高。②高氧对照组MMPs、TIMPs mRNA表达和MMP-2/TIMP-2、MMP-9/TIMP-1比值较空气对照组明显增高。③地塞米松能剂量依赖性下调MMP-2、MMP-9 mRNA表达;对TIMP-1、TIMP-2 mRNA表达有一定程度的抑制效应;随地塞米松浓度的增加,MMP-2/TIMP-2、MMP-9/TIMP-1比值亦逐渐降低。结论 地塞米松下调MMPs mRNA表达,调节MMPs/TIMPs之间的失衡,可能是其减轻高氧肺损伤的机制之一。     

心肌肥厚大鼠细胞外基质重塑中基质金属蛋白酶的作用及氯沙坦干预的效果

背景细胞外基质中的纤维性胶原过度沉积是心肌肥厚发展过程中的重要特征.基质金属蛋白酶(matrix metalloproteinases,MMPs)及其生理性抑制剂(tissue inhibitor of metalloproteinases,TIMPs)是调节细胞外基质的重要酶类,转化生长因子β1(transforming growthfactor-β, TGF-β1)能促进组织纤维化.目的探讨MMP-9及其生理性抑制剂TIMP-1和TGF-β1在心肌肥厚大鼠细胞外基质重塑中的作用及氯沙坦干预的效果.设计随机对照实验研究.地点、材料和干预南京医科大学动物实验中心雄性SD大鼠随机用随机数字表法分为3组对照组、去甲肾上腺素组[1.06 mg/(kg·d)×15 d]、去甲肾上腺素+氯沙坦组[10 mg/(kg.d)×15 d].去甲肾上腺素腹腔注射,2次/d,连续15d,建立心肌肥厚的模型.氯沙坦灌胃给药.主要观察指标腹腔注射生理盐水、去甲肾上腺素、氯沙坦灌胃给药大鼠超声心动图检查,病理检查,心肌胶原蛋白表达,MMP-9,TIMP-1和TGF-β1mRNA和蛋白表达情况的对比结果.结果大鼠腹腔注射去甲肾上腺素后发生左心室肥厚及纤维化,胶原的含量及MMP-9(82.9±18.3),TIMP-1(43.8±7.3)和TGF-β1(49.4±7.9),mRNA的表达显著高于健康对照组.氯沙坦能降低室间隔的厚度,减少左室心肌中总体胶原、Ⅰ型、Ⅲ型胶原的合成及MMP-9,TGF-β1的表达.结论MMP-9,TIMP-1和TGF-β1与去甲肾上腺素诱导的心肌细胞外基质重塑有关.氯沙坦能有效的防治心肌肥厚及细胞外基质重塑,这一效应与其降低心肌中高表达的MMP-9和TGF-β1有关.     

Circulating matrix metalloproteinases and their inhibitors in premature coronary atherosclerosis.

To investigate the clinical significance of circulating matrix metalloproteinases (MMPs) and their tissue inhibitos (TIMPs) in patients with premature coronary atheroscrelosis, we studied 53 consecutive male patients with angiographically defined premature (<65 years) and stable coronary artery disease. Plasma levels of MMP-2, MMP-3, MMP-9, TIMP-1, and TIMP-2 were determined in peripheral blood by a sandwich enzyme immunoassay, and the results were compared with those from 133 age-matched control males. There were significant differences in all the MMPs and TIMPs (p<0.001) between patients and controls. In the patient group, the levels of MMP-9 (mean +/- SD (ng/ml) 27.2 +/- 15.2/21.8 +/- 15.2) and TIMP-1 (130.4 +/- 55.7/94.5 +/- 26.3) were significantly higher, and the levels of MMP-2 (632.5 +/- 191.6/727.6 +/- 171.4), MMP-3 (53.1 +/- 31.2/79.6 +/- 29.9), and TIMP-2 (24.7 +/- 15.2/35.4 +/- 16.4) were significantly lower than those of controls. We found significant positive correlation between plasma MMP-9 levels and low-density lipoprotein (LDL)-cholesterol levels (Rs = 0.168, p = 0.022), and significant negative correlation between plasma MMP-9 levels and high-density lipoprotein (HDL)-cholesterol levels (Rs = -0.164, p = 0.026) by Spearman rank correlation test. In contrast, plasma MMP-2 (Rs = 0.181, p = 0.014) and MMP-3 (Rs = 0.260, p = 0.0004) levels were positively correlated with HDL-cholesterol levels. TIMP-2 levels were negatively correlated with total cholesterol (Rs = -0.197, p = 0.007) and LDL-cholesterol (Rs = -0.168, p=0.022) levels. These results suggest that the circulating levels of MMPs and TIMPs are altered in patients with premature coronary atherosclerosis and that plasma lipoprotein cholesterol levels correlate with these, possibly as a result of the lipoprotein-vessel wall interactions.     

Relationship between matrix metalloproteinases/tissue inhibitors of matrix metalloproteinases systems and autoantibody patterns in systemic sclerosis

OBJECTIVE: Systemic sclerosis (SSc) is a connective tissue disease characterized by tissue fibrosis that reflects an imbalance between collagen production and degradation. Matrix metalloproteinases (MMPs) are a family of endopeptidases involved in the remodelling of extracellular matrix (ECM). This activity is controlled by tissue inhibitors of MMP (TIMPs). Aim of this study was the evaluation of MMP-9/TIMP-1 and MMP-2/TIMP-2 systems in patients with SSc. DESIGN AND METHODS: SearchLight Human MMP Array 1 was used to measure MMPs and TIMPs in 32 SSc patients and 32 matched healthy controls. RESULTS: SSc patients showed higher values of both MMP-9 and TIMP-1 in comparison with controls. The patients with anticentromere antibodies (ACA) positivity showed higher values of MMPs and TIMPs in comparison with either controls or the patients with anti-Scl70-positive antibodies. CONCLUSION: Results of this investigation suggest that SSc patients with ACA positivity, after a primary fibrogenetic noxa, react with a more abundant release of MMP/TIMP, whereas patients with anti-Scl70 antibody show a normal response.     

缬沙坦对人脐动脉平滑肌细胞MMP-9、TIMP-1表达的影响——附106份检测报告

目的：探讨缬沙坦对人脐动脉平滑肌细胞基质金属蛋白酶-9（matrix metal—loproteinase,MMP-9）、组织型金属蛋白酶抑制物-1（tissue inhibitor of metalloproteinase,TIMP-1）表达的影响。方法：对人脐动脉平滑肌细胞进行原代培养,根据平滑肌原代细胞的培养方式分为氧化低密度脂蛋白（oxidized low—density lipoprotein,ox-LDL）培养组（ox-LDL组）、AngⅡ加ox-LDL培养组（AngⅡ组）、AngⅡ加ox-LDL加缬沙坦培养组（缬沙坦组）、普通培养基培养组（对照组）,应用ELISA法及逆转录PCR法分别检测4组上清液中的MMP-9、TIMP—1含量,以及细胞内MMP-9、TIMP-1 mRNA的相对表达量,并应用统计学方法进行比较。结果：AngⅡ组上清液中的MMP-9含量均明显高于其他3组（均为P〈0．01）。ox-LDL组与AngⅡ组上清液中TIMP-1的含量均明显低于对照组（均为P〈0．01）,缬沙坦组与对照组上清液中TIMP-1的含量比较差异无统计学意义（P〉0．05）。另外,AngⅡ组细胞内MMP-9 mRNA的相对表达量均明显高于其他3组（均为P〈0．01）,其TIMP-1 mRNA的相对表达量均明显低于对照组、ox-LDL组及缬沙坦组（均为P〈0．01）,缬沙坦组细胞内TIMP-1 mRNA的相对表达量明显低于对照组（P〈0．05）,但与ox-LDL组比较差异无统计学意义（P〉0．05）。结论：AngⅡ可增加人脐动脉平滑肌细胞MMP-9的表达而降低TIMP-1的表达,缬沙坦可以抑制AngⅡ的这种作用。     

Dynamic changes of matrix metalloproteinases and their tissue inhibitors in severe sepsis

Purpose

Little is known about the dynamic changes of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) in sepsis. Our aim was therefore to investigate the time course of MMPs and their inhibitors in patients experiencing severe sepsis.

Methods

Our prospective controlled analysis included 38 patients with severe sepsis. Plasma levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 were measured daily at a 5-day-long period with enzyme-linked immunosorbent assay. Seventeen healthy volunteers were invited as controls.

Results

MMP-2 showed no difference compared to controls, whereas significantly elevated MMP-9 levels were detected on admission (P < .005). Significantly elevated but declining TIMP-1 levels were measured during the whole trial (P < .002-.004). Except for the second day, TIMP-2 levels were significantly lower than controls (P < .05-.009). MMP2/TIMP-1 ratios were significantly lower in septic patients (P < .03-.006), whereas MMP-2/TIMP-2 ratios were elevated throughout our study (P < .03-.006). MMP-9/TIMP-1 ratios were significantly lower at the first 3 days (P < .05-.008). MMP-9/TIMP-2 was significantly elevated on admission (P < .006).

Conclusions

Our research is the first follow-up study dealing with MMPs, TIMPs, and their ratios in severe sepsis. Our results indicate that MMPs and TIMPs may play a crucial role in severe sepsis, especially TIMP-1, MMP-9, and possibly TIMP-2, after an extensive study.     

MMP-3、MMP-9和TIMP-1的动态平衡可作为心肌基质重构的重要标志

目的从基质金属蛋白酶（matrix metalloproteinase, MMPs）和基质金属蛋白酶组织抑制剂（tissue inhibitor of metalloproteinase, TIMps）的动态平衡着手探寻其对心肌基质的组织修复和再生作用,为心肌损伤后基质重构造成的心肌肥大、增厚等引起的心力衰竭提供的有效的生物检测方法及治疗。方法利用广州市红十字会医院（暨南大学第四附属医院）的心脏病患者作为研究对象,按照随机区组分组的方法根据进行的心脏手术不同,将研究对象分为3个组,第1组为先天性心脏病组（CHD组）;第2组为风湿性心脏病组（PHD组）;第3组为冠心病组（COR组）,由于其房室大小,心功能正常,设其为对照组。Elisa法对患者术前血液中的mmps和timps进行定量测定,寻求它们之间的动态平衡关系;术中切取心肌组织,用免疫组化方法测定心肌基质的胶原形态和分布;rt-PCR技术对心肌组织中mmps和timps的mRNA进行检测,查看其mRNA表达情况。结果CHD组、PHD组术前血液中的MMP-3、MMP-9、TIMP-1含量均较COR组高,MMP-9增多更为显著,差异有统计学意义（P < 0.05）,而CHD组和PHD组差异不显著,无统计学意义;免疫组化结果显示,CHD组、PHD组中MMP-3、MMP-9、TIMP-1均较COR组分布广泛,PCR结果显示CHD组、PHD组患者MMP-1、MMP-9、TIMP-1的mRNA表达较COR组升高,差异有统计学意义（P < 0.05）,同样CHD组和PHD组差异不显著,无统计学意义。结论MMP-1、MMP-9和TIMP-1的动态平衡可作为心肌基质重构的重要标志,调节MMPs和TIMPs的活性有望成为心脏病治疗的新方向。     

Increase in the ratio of matrix metalloproteinase-9 to tissue inhibitor of metalloproteinase-1 in saliva from patients with primary Sjögren's syndrome

BACKGROUND: To investigate the ratio of matrix metalloproteinase (MMP) to tissue inhibitor of metalloproteinase (TIMP) in primary Sj?gren's syndrome (PSS), patients and healthy subjects MMP-2, 9 and TIMP-1, 2 levels were measured in saliva. METHODS: Stimulated whole-mixed saliva was collected from 32 patients and 26 healthy subjects. MMP-2, 9 and TIMP-1, 2 levels were measured using enzyme-linked immunosorbent assay (ELISA) and the sandwich enzyme immunoassay (sandwich EIA). Zymography and reverse zymography were used to identify MMPs and TIMPs. RESULTS: MMP-9 (gelatinase-B) level in saliva was significantly increased in the patients. MMP-9 (ng/ml): patients 231.02 +/- 151.77 (mean +/- S.D.), healthy subjects 145.87 +/- 111.65 (p < 0.05). MMP-2 levels were not detected with this system kit in either healthy subjects or patients. The differences in TIMPs were only trends and not statistically significant (p > 0.05). Accordingly, MMP-9/TIMP-1 was greatly increased in the patients (2.60 +/- 1.18) than in the healthy subjects (1.28 +/- 1.11) (p < 0.01). CONCLUSION: This study found that MMP-9/TIMP-1 and MMP-9 levels in the saliva were significantly higher in pSS patients than those in healthy subjects. Our results indicate that the increase in MMP-9/TIMP-1, rather than the increase in MMP-9, in pSS patients' saliva is strongly involved in destruction of glandular and salivary duct tissues.     

Angiotensin II stimulates extracellular matrix protein synthesis through induction of transforming growth factor-beta expression in rat glomerular mesangial cells.

Angiotensin II (Ang II) has been implicated in the development of progressive glomerulosclerosis, but the precise mechanism of this effect remains unclear. In an experimental model, we have shown previously that TGF-beta plays a key role in glomerulosclerosis by stimulating extracellular matrix protein synthesis, increasing matrix protein receptors, and altering protease/protease-inhibitor balance, thereby inhibiting matrix degradation. We hypothesized that Ang II contributes to glomerulosclerosis through induction of TGF-beta. Ang II treatment of rat mesangial cells in culture increased TGF-beta and matrix components biglycan, fibronectin, and collagen type I at both the mRNA and protein levels in a time- and dose-dependent manner. Saralasin, a competitive inhibitor of Ang II, prevented the stimulation. Ang II also promoted conversion of latent TGF-beta to the biologically active form. Coincubation of mesangial cells with Ang II and neutralizing antibody to TGF-beta blocked the Ang II-induced increases in matrix protein expression. Continuous in vivo administration of Ang II to normal rats for 7 d resulted in 70% increases in glomerular mRNA for both TGF-beta and collagen type I. These results indicate that Ang II induces mesangial cell synthesis of matrix proteins and show that these effects are mediated by Ang II induction of TGF-beta expression. This mechanism may well contribute to glomerulosclerosis in vivo.     

血清基质金属蛋白酶及其组织抑制物的变化及临床意义

目的探讨原发性高血压（EH）患者血清基质金属蛋白酶（MMPs）及其组织抑制物（TIMPs）检测的变化及临床意义。方法采用酶联免疫吸附试验法分别测定90例未经治疗的EH患者（EH组）和60例健康体检者（对照组）血清MMP-9和TIMP-1水平。经6个月降压治疗后，检测EH患者的血清MMP-9和TIMP-1水平，并与治疗前进行对比分析。结果治疗前EH组的血清MMP-9和TIMP-1水平明显低于对照组，差异有统计学意义（P〈0、01，t=8、484、8、483）。EH组患者MMP-9与TIMP-1呈显著正相关（r=0．585，P〈0、01），但其水平与收缩压和舒张压无相关性（P〉0．05）。与治疗前相比，EH组治疗后的血清MMP-9和TIMP-1水平明显增高，差异有统计学意义（P〈0．01，t=16．597、26．298）。结论EH患者MMP-9和TIMP-1降低可能反映了EH患者细胞外基质代谢异常，可将其作为EH相关血管活性异常的间接标志物。     

基质金属蛋白酶及其抑制物对葡萄胎恶变的预测价值研究

目的　探讨基质金属蛋白酶 (MMPs)及其抑制物TIMPs在葡萄胎恶变预测中的价值。方法　采用免疫组化SP法测定 39例葡萄胎 (其中 11例发生恶变 )和 17例正常相同孕周的绒毛组织中MMP 2、MMP 9、TIMP 1、TIMP 2的表达及分布。结果　正常绒毛组织、葡萄胎组织的滋养细胞胞浆中MMP 2、MMP 9、TIMP 1、TIMP 2均有表达。MMP 2、MMP 9在葡萄胎发生恶变组的表达明显强于正常绒毛组和葡萄胎未发生恶变组 ,而TIMP 1,TIMP 2在正常绒毛组和葡萄胎组之间表达无显著性差异。MMP 2 /TIMP 2、MMP 9/TIMP 1在葡萄胎恶变组织中的表达强于在正常绒毛组织的表达。结论　在葡萄胎清宫组织中进行MMP 2、MMP 9的检测 ,有望成为判断葡萄胎日后恶变的一项预测指标     

Elevated circulating levels of matrix metalloproteinase-9 in type 1 diabetic patients with and without retinopathy

OBJECTIVE: Tissue expression pattern of matrix metalloproteinases (MMPs) and their inhibitors TIMPs indicate that microvascular complications of diabetes mellitus are associated with extracellular matrix remodelling. We investigated whether circulating levels of MMP-9 and TIMP-1 are altered in diabetic retinopathy and whether they might serve as biological markers of ocular complications in type 1 diabetes. RESEARCH DESIGN AND METHODS: We recruited 47 type 1 diabetic patients free of vascular complications (n=40) or with retinopathy (n=14). Patients with macroangiopathy, neuropathy and nephropathy were excluded. A group of nondiabetic control subjects (n=35) was also constituted for comparative purposes. Peripheral blood levels of MMP-9 and TIMP-1 were determined using immunoenzymatic assays. RESULTS: Type 1 diabetic subjects exhibited significantly higher circulating levels of both MMP-9 and MMP-9/TIMP-1 ratio, as well as a tendency to increased serum TIMP-1 levels relative to nondiabetic controls (p<0.001). Diabetic patients with retinopathy also displayed elevated systemic values of MMP-9 and MMP-9/TIMP-1 ratio when compared to patients without retinopathy (p<0.05). Logistic regression analysis identified diabetes duration firstly (P<0.01), and MMP-9 serum levels secondly (P<0.01) as significant and independent variables associated with the existence of retinopathy. CONCLUSIONS: Our data suggest that peripheral blood MMP-9 levels might serve as surrogate biomarkers of retinopathy in type 1 diabetic patients free of other vascular complications.     

慢性充血性心力衰竭患者血清基质金属蛋白酶与骨桥蛋白变化的相关性研究

目的探讨慢性充血性心力衰竭（CHF）患者血清基质金属蛋白酶（MMPs）及其组织抑制因子（TIMPs）和骨桥蛋白（OPN）的变化及相关性。方法CHF患儿24例，正常对照组15例。采用ELISA法测定血清MMP-9和TIMP-1的含量,ELISA法测定血清OPN的含量，分析MMP-9／TIMP-1值与OPN和左室舒末内径（LVEDD）及射血分数（EF）的关系。结果CHF患者血清MMP-9、MMP-9／TIMP-1值和OPN含量明显增加（P均〈0．01），TIMP-1含量明显减少（P〈0．01），MMP-9／TIMP-1值与OPN含量和LVEDD呈正相关（P〈0．05），与EF呈负相关（P〈0．05）。结论CHF患者MMP-9／TIMP-1值与OPN含量呈明显的正相关；MMP-9／TIMP-1值与OPN含量的变化，可反映心衰的严重程度并加速了心功能的恶化。     

Matrix metalloproteinases and their inhibitors: promising novel biomarkers in severe sepsis?

The multicenter study conducted by Lorente and coworkers published in the previous issue of Critical Care demonstrates that matrix metalloproteinase (MMP)-9 and MMP-10 and their inhibitor tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) are promising novel biomarkers to predict severity and outcome of sepsis. In recent years MMPs have emerged as biomarkers in a variety of diseases, such as sepsis, coronary artery disease, cancer, heart failure, chronic lung disease and rheumatoid arthritis. MMPs constitute a family of proteinases that are expressed during developmental, physiological, and pathophysiological processes, for example as a response to infection. Excessive inflammation following infection may cause tissue damage, and MMPs are implicated in causing this immunopathology. The activity of MMPs is regulated by secretion of specific inhibitors (TIMPs). Studies using MMP inhibitors and MMP knockout mice indicate that MMPs play an essential role in infection and in the host response to infection. The measurement of MMP-9 and MMP-10 and their inhibitor TIMP-1 in the intensive care setting could be an attractive noninvasive tool for determination of outcome of septic patients.     

R(+)-methanandamide and other cannabinoids induce the expression of cyclooxygenase-2 and matrix metalloproteinases in human nonpigmented ciliary epithelial cells

Prostaglandins (PGs) and matrix metalloproteinases (MMP) have been implicated in lowering intraocular pressure (IOP) by facilitating aqueous humor outflow. A possible role of cyclooxygenase-2 (COX-2) in this process was emphasized by findings showing an impaired COX-2 expression in the nonpigmented ciliary epithelium (NPE) of patients with primary open-angle glaucoma. Using human NPE cells, the present study therefore investigated the effect of the IOP-lowering cannabinoid R(+)-methanandamide [R(+)-MA] on the expression of COX-2 and different MMPs and tissue inhibitors of MMPs (TIMPs). R(+)-MA led to a concentration- and time-dependent increase of COX-2 mRNA expression. R(+)-MA-induced COX-2 expression was accompanied by time-dependent phosphorylations of p38 mitogen-activated protein kinase (MAPK) and p42/44 MAPK and was abrogated by inhibitors of both pathways. Moreover, R(+)-MA increased the mRNA and protein expression of MMP-1, MMP-3, MMP-9, and TIMP-1 but not that of MMP-2 and TIMP-2. Inhibition of COX-2 activity with NS-398 [N-[2-(cyclohexyloxy)-4-nitrophenyl]-methanesulfonamide] was associated with a virtually complete suppression of R(+)-MA-induced MMP-9 and TIMP-1 expression. Consistent with these data, MMP-9 and TIMP-1 expression was also induced by PGE2, a major COX-2 product. Two other COX-2-inducing cannabinoids, anandamide and Delta9-tetrahydrocannabinol, caused the same pattern of MMP and TIMP expression as R(+)-MA both in the absence and presence of NS-398. Altogether, cannabinoids induce the production of several outflow-facilitating mediators in the human NPE. Our results further imply an involvement of COX-2-dependent PGs in MMP-9 and TIMP-1 expression. In conclusion, stimulation of intraocular COX-2 and MMP expression may represent a potential mechanism contributing to the IOP-lowering action of different cannabinoids.     

hCG对滋养细胞系侵袭性相关基因表达的影响

目的：揭示人类绒毛膜促性腺激素（hCG）对滋养细胞侵袭性的影响。 方法：以永生化的滋养细胞系JEG-3为研究对象，采用逆转录多聚酶链反应（RT-PCR）方法，观察了hCG对滋养细胞系侵袭性相关基因表达的影响。 结果：JEG-3细胞自身表达MMP-2，而不表达MMP-9或uPA，同时还表达这些蛋白酶的抑制因子TIMP-1、TIMP-2和PAI-1。JEG-3细胞还表达组织蛋白酶B，而未见组织蛋白酶D的表达。在与25 IU/ml hCG温育50h后，MMP-2的表达无显著变化，而TIMP-1和PAI-1的表达显著降低，同时TIMP-2的表达被诱导增加，其它基因的表达无明显变化。 结论：高浓度(25 IU/ml)的hCG可能通过改变蛋白水解酶及其抑制因子的表达而减弱滋养细胞的侵袭性。     

螺内酯对肝星状细胞表达基质金属蛋白酶及其抑制剂的影响

目的研究螺内酯干预肝星状细胞（HSCs）后基质金属蛋白酶2（MMP-2）、MMP-9、MMP-13和其组织抑制剂1（TIMP-1）的变化。探讨螺内酯对胶原代谢影响的机制。方法应用不同浓度螺内酯（1×10^-4）mol／L-（1×10^-7）mol／L干预HSCs 48小时,采用反转录-聚合酶链反应（RT-PCR）方法检测MMP-2、MMP-9、MMP-13 mRNA和TIMP-1 mRNA的表达。结果①螺内酯组的MMP-13基因表达强度上调,螺内酯（1×10^-4）mol／L-（1×10^-5）mol／L浓度组MMP-13基因表达强度（0．91±0．13、0．80±0．01、0．67±0．15）均明显高于对照组（0．53±0．10）（P〈0．01）。1×10^-4mol／L浓度组MMP-13基因表达强度接近对照组的2倍。②螺内酯干预HSCs48小时后,TIMP-1基因表达减少,螺内酯（1×10^-4）mol／L-（1×10^-6）mol／L浓度组（0．15±0．05、0．28±0．15、0．37±0．03）明显低于对照组（0．47±0．04）（P〈0．01或〈0．05）。③螺内酯不同浓度干预HSCs 48小时后,HSCs MMP-2、MMP-9基因表达无明显变化（P〉0．05）。结论螺内酯可抑制HSCs TIMP-1基因的表达,增加间质胶原酶MMP-13 mRNA的含量。螺内酯可能通过对MMPs及TIMP-1影响而促进胶原降解。