Matrix Metalloproteinase-2 and Tissue Inhibitor of Metalloproteinase-1 in the Retinal Pigment Epithelium and Interphotoreceptor Matrix: Vectorial Secretion and Regulation

Matrix metalloproteinases (MMPs) and the tissue inhibitors of metalloproteinases (TIMPs) play an essential role in both normal and pathological extracellular matrix degradation, and a TIMP has been associated with at least one type of retinal degeneration. We have studied expression of MMP-2 and TIMP-1 by zymography, immunocytochemistry, and immunoblotting in the retinal pigment epithelium (RPE) from normal, aged and diseased retinas. MMPs and TIMPs were found in the rat RPE, interphotoreceptor matrix (IPM), and in media conditioned by human and rat RPE in culture. In other polarized cells, MMPs and TIMP-2 are secreted vectorially towards the basal lamina. In the RPE, however, MMP-2 and TIMP-1 were secreted preferentially from the apical surface, the surface bordering the IPM. These findings provide new evidence that MMPs and TIMPs could play a role in the turnover of IPM components.Cell homogenates and conditioned media from RPE isolated from mutant Royal College of Surgeons (RCS) rats with inherited retinal dystrophy had similar amounts of MMP-2 and TIMP-1 as those from congenic control rats. The secretion of MMP-2 and TIMP-1 from RPE cell cultures isolated from young and aged human donors varied widely. However, with increasing cell passage number, secretion of MMPs and TIMPs from human RPE increased dramatically. Also, growing human RPE on bovine corneal endothelial cell-generated extracellular matrix instead of plastic reduced the secretion of both MMPs and TIMPs. These data suggest that the integrity of Bruch's membrane may serve to regulate RPE functions in MMP and TIMP secretion and that extracellular matrices contain signals that regulate MMP and TIMP synthesis and/or secretion by the RPE.     

Expression of gelatinases within the trabecular bone compartment of ovariectomized and parathyroidectomized adult female rats

Ovariectomized (ovx) and parathyroidectomized (ptx) rat models of disturbed bone metabolism have been widely used in evaluating bone changes resulting from hormonal depletion, and are characterized by elevated and depressed bone turnover, respectively. We report here the expression of gelatinases extracted from native trabecular bone in these models. Nine-month-old female Sprague-Dawley rats were sacrificed after 3 weeks following ovx or 10 days post ptx to determine the influence of these procedures on the levels of proximal tibial bone tissue gelatinases. Identification and quantitation of these enzymes were performed via gelatin gel zymography of native tissue extracts and laser densitometry of developed gels, respectively. In the ptx model, a reduction in tissue levels of pro- and active-MMP-2 and 45 kDa activated fragment was seen, whereas ovx exhibited significant increases in these enzymes. The MMPs are therefore clearly under the influence of factors known to modulate bone remodeling in vivo. The study of MMP levels directly extracted from bone using these experimental models may assist in developing management regimes for metabolic bone diseases through the use of drugs aimed at controlling turnover.     

Expression of matrix metalloproteinase matrilysin (mmp-7) was induced by activated ki-ras via ap-1 activation in sw1417 colon cancer cells

The matrix metalloproteinase matrilysin (MMP-7) is a member of the matrix metallo-proteinase gene family, which is believed to play an important role in tumor invasion and metastasis. We have previously found that matrilysin mRNA is specifically expressed in colorectal cancers and adenomas and that its message is localized in the tumor cells themselves. We examined the effects of activated Ki-ras oncogene on the expression of matrilysin in colon cancer cells. We showed that both mRNA and the enzymatic activity of matrilysin were induced by the introduction of activated Ki-ras into SW1417 colon cancer cells. To understand the mechanisms regulating this induction, we analyzed alterations of AP-1 activity induced by activated Ki-ras, using the chloramphenicol acetyltransferase assay. AP-1 activity in SW1417 cells expressing activated Ki-ras was higher than that in control cells. The gel-shift assay also showed higher levels of AP-1 binding protein in SW1417 cells expressing activated Ki-ras than those in control cells. Our results suggest that activated Ki-ras may play a role in inducing expression of matrilysin through an AP-1-dependent pathway in colon cancer cells.     

高血压大鼠心肌中MMP-2的蛋白表达及其RAS阻断后的变化

[目的]探讨高血压大鼠左室心肌中基质金属蛋白酶-2(MMP-2)蛋白表达的变化以及血管紧张素转化酶抑制剂(ACEI)、血管紧张素Ⅱ1型受体拮抗剂(AT1-ant)单独与联合治疗对高血压大鼠心肌中MMP-2蛋白表达的影响。[方法]40只雄性8周龄的易卒中自发性高血压大鼠(SHRSP)随机分成5组:SHRSP对照组、安慰剂组、缬沙坦组、苯那普利组及缬沙坦与苯那普利联用组。另外,取8只雄性8周龄的京都Wistar大鼠(WKY)作为对照。利用免疫组织化学的方法检测WKY以及SHRSP左室心肌中MMP-2蛋白的表达。[结果]SHRSP的收缩压(SBP)、左室质量指数(LVMI)、胶原容积分数(CVF)、血管周围胶原面积(PVCA)、左室心肌MMP-2蛋白表达较同龄的WKY显著增高。给予苯那普利、缬沙坦单独或联合治疗后SHRSP的LVMI、CVF、PVCA、左室心肌 MMP-2蛋白表达都显著降低。苯那普利、缬沙坦单独应用时的效果没有差异,而联合应用的效果更显著。[结论]SHRSP左室心肌中MMP-2蛋白表达增高,肾素血管紧张素系统(RAS)阻断后MMP-2蛋白表达显著减少;苯那普利、缬沙坦逆转高血压心室重塑的作用可能部分通过下调MMP-2蛋白表达而实现;苯那普利和缬沙坦都能降低SHRSP的血压以及逆转其左室重塑,而联合用药的作用更显著。     

糖尿病大鼠肺病理改变及同期肾脏病理变化对比

目的:观察糖尿病(DM)大鼠肺组织改变及与同期肾脏变化的关系.方法:链脲菌素腹腔注射制作糖尿病大鼠模型,4周后胶原、网状纤维染色及透射电镜方法观察糖尿病大鼠肺组织基底膜病理改变,同期观察肾脏改变.结果:DM大鼠4周后肺组织病理改变为毛细血管基底膜及Ⅱ型肺泡上皮细胞基底膜不同程度的增厚及肺间质胶原成分等细胞外基质的增多,与同期糖尿病肾脏病变相平行.结论:DM大鼠4周后肺组织与糖尿病肾病相似,主要表现为微血管病变.     

MMP-2和Kiss-1在膀胱癌中的表达及意义

目的:研究基质金属蛋白酶2(MMP-2)和肿瘤转移抑制因子Kiss-1在膀胱癌中的表达以及与肿瘤侵袭的关系.方法:采用免疫组织化学S-P法检测40例膀胱癌和10例正常膀胱组织标本中MMP-2和Kiss-1的表达,并与临床病理参数进行比较.结果:膀胱癌MMP-2阳性率57.5%,表达与肿瘤分期、分级密切相关(P＜0.05).Kiss1阳性率为42.5%,表达随肿瘤分期、分级升高呈下降趋势.MMP-2与Kiss-1的表达无显著相关性(P＞0.05).结论:MMP-2和Kiss-1的表达水平对于评价膀胱癌的预后具有重要意义.     

Experimental Study on Allogenic Decalcified Bone Matrix as Carrier for Bone Tissue Engineering

The biocompatibility and osteogenic activity of allogenic decalcified bone matrix (DBM) used as a carrier for bone tissue engineering were studied. Following the method described by Urist, allogenic DBM was made. In vitro, DBM and bone marrow stromal cell (BMSC) from rab-bits were co-cultured for 3-7 days and subjected to HE staining, and a series of histomorphological observations were performed under phase-contrast microscopy and scanning electron microscopy (SEM). In vivo the mixture of DBM/BMSC co-cultured for 3 days was planted into one side of muscules sacrospinalis of rabbits, and the DBM without BMSC was planted into other side as con-trol. Specimens were collected at postoperative week 1, 2 and 4, and subjected to HE staining, and observed under SEM. The results showed during culture in vitro, the BMSCs adherent to the wall of DBM grew, proliferated and had secretive activity. The in vivo experiment revealed that BMSCs and undifferentiated mesenchymal cells in the perivascular region invaded gradually and proliferated together in DBM/BMSC group, and colony-forming units of chondrocytes were found. Osteoblasts,trabecular bone and medullary cavity appeared. The inflammatory reaction around muscles almost disappeared at the second weeks. In pure DBM group, the similar changes appeared from the sur-face of the DBM to center, and the volume of total regenerate bones was less than the DBM/BMSC group at the same time. The results indicated that the mixture of DBM and BMSC had good bio-compatibility and ectopic induced osteogentic activty.     

Cell–matrix adhesion in vascular development

Summary.  Vascular development requires correct interactions among endothelial cells, pericytes and surrounding cells. These interactions involve many cell adhesion interactions, including cell–matrix interactions both with basement membranes and with surrounding extracellular matrices. Investigations of the contributions of these various interactions in vascular development and angiogenesis have been rather uneven and incomplete over the past 10–15 years. There has been considerable concentration on a few receptors, matrix proteins and proteolytic fragments with the goal of finding means to control angiogenesis. Many other potential contributors have received much less attention. Even for those molecules that have been subject to intensive investigation, our knowledge is incomplete. This review will survey the spectrum of extracellular matrix (ECM) proteins and cell–matrix adhesion receptors (particularly integrins) that are likely to contribute to angiogenesis and discuss what is known and not known about the roles of each of them.