腹主动脉瘤发病机理的实验研究 Ⅱ.基质金属蛋白酶-9在大鼠腹主动脉瘤模型中的动态表达

目的 :检测MMP 9在大鼠正常动脉及动脉瘤模型组织中的动态表达 ,以探讨其在腹主动脉瘤发病机制中的作用。方法 :Wistar大鼠 5 4只 ,随机均分为 1组正常对照组、4组灌注对照组和 4组实验组。实验组腹主动脉灌注弹力蛋白酶 (2 5U ml) 2ml,灌注对照组灌注生理盐水 2ml,分别于术前、术后即刻、2d、7d、14d测量腹主动脉直径 ,并采用免疫组织化学和分子原位杂交技术动态检测腹主动脉组织中MMP 9的表达。结果 :正常及灌注对照组腹主动脉组织中均未发现MMP 9,而实验组弹力蛋白酶灌注后2～ 14dMMP 9均有不同程度升高 ,第 7d达到高峰 ,第 14d有所回落。结论 :MMP 9分泌的增加可能与炎症反应有关 ,且为腹主动脉瘤形成中不可或缺的一步。     

Ⅱ.基质金属蛋白酶-9在大鼠腹主动脉瘤模型中的动态表达

目的：检测MMP-9在大鼠正常动脉及动脉瘤模型组织中的动态表达，以探讨其在腹主动脉瘤发病机制中的作用。方法：wistar大鼠54只，随机均分为1组正常对照组、4组灌注对照组和4组实验组。实验组腹主动脉灌注弹力蛋白酶(25U／ml)2ml，灌注对照组灌注生理盐水2ml，分别于术前、术后即刻、2d、7d、14d测量腹主动脉盲径，并采用免疫组织化学和分子原位杂交技术动态检测腹主动脉组织中MMP-9的表达。结果：正常及灌注对照组腹主动脉组织中均未发现MMP-9，而实验组弹力蛋白酶灌注后2-14d MMP-9均有不同程度升高，第7d达到高峰，第14d有所回落。结论：MMP-9分泌的增加可能与炎症反应有关，且为腹主动脉瘤形成中不可或缺的一步。     

基质金属蛋白酶9在胸主动脉瘤和胸主动脉夹层中的表达

目的 观察基质金属蛋白酶(MMP)-9在DebakeyI型胸主动脉夹层(TAD)和胸主动脉瘤(TAA)中的表达,探讨其在TAD和TAA中的作用.方法 苏木素-伊红(HE)染色、铁苏木素染色、TUNEL染色、免疫组织化学染色分别观察11例TAD和10例TAA的病理特征、弹力纤维断裂、管壁细胞凋亡、MMP-9在动脉管壁中的表达和定位,并分析MMP-9表达与各项临床参数的关系.结果 MMP-9在TAD和TAA中高表达(P<0.05),主要位于中外膜平滑肌细胞和炎性细胞,MMP-9表达与主动脉直径显著相关(P<0.05).结论 MMP-9在TAD和TAA中高表达,中外膜平滑肌细胞和炎性细胞是其主要表达细胞;MMP-9可能通过弹力板层的破坏来影响管壁重构过程.     

Cigarette smoking increases the risk of developing a cerebral aneurysm and of subarachnoid hemorrhage]

Despite recent advances in diagnostic and therapeutic techniques, subarachnoid hemorrhage (SAH) is still a serious condition associated with high mortality and morbidity. There are no effective treatments other than surgical intervention. However, another option for decreasing the occurrence of SAH may be prevention of aneurysms formation and of their rupture by controlling risk factors. Cigarette smoking has been recently shown to be one of the major risk factors for SAH. We investigated whether cigarette smoking increased the risk of developing cerebral aneurysms and of SAH. Degree of smoking was investigated in 182 patients with SAH and in 123 patients with an unruptured cerebral aneurysm incidentally detected during investigation of other diseases. Sixty-nine patients with other diseases who were shown to be free of cerebral aneurysms through MR angiography served as controls. Smoking significantly increased the risk of both aneurysm formation and SAH; The odds ratio for SAH was 2.4, and for unruptured cerebral aneurysm 1.7. Smoking especially increased the occurrence of SAH in women and in youngsters. However, smoking did not influence the occurrence of cerebral vasospasm and multiplicity of aneurysms. These data suggest the importance of avoiding smoking to prevent the occurrence of cerebral aneurysms and of SAH.     

罗格列酮对肝星状细胞基质金属蛋白酶-2和-9基因表达的影响

目的 观察过氧化物酶体增殖物活化受体γ(PPARγ)特异配体罗格列酮对肝星状细胞(HSC)表达基质金属蛋白酶(MMP)-2和MMP-9的影响.方法 在培养的HSC株中加入不同浓度的罗格列酮(终质量浓度为5、10、15、20 μmol/L),于24 h后收取细胞,利用半定量逆转录-聚合酶链反应(RT-PCR)测定MMP-2、MMP-9 mRNA表达.结果 MMP-2表达水平在罗格列酮5、10、15、20 μmol/L组分别为0.5708±0.0609、0.8900±0.0823、1.1348±0.1205、1.4490±0.0832,而空白对照组为0.3237±0.0796.MMP-9表达水平在罗格列酮5、10、15、20 μmol/L组分别为0.5487±0.0770、0.7554±0.0510、0.9497±0.0451、1.1088±0.0777,空白对照组为0.3220±0.0592.结论 罗格列酮可增强HSC对MMP-2、MMP-9的表达,并在一定范围内,呈剂量依赖性.     

Response of plasma matrix metalloproteinase-9 to conventional abdominal aortic aneurysm repair or endovascular exclusion: implications for endoleak

PURPOSE: Matrix metalloproteinases are enzymes capable of breaking down all of the components of the extracellular matrix and have been implicated in the development of aneurysm formation. Because matrix metalloproteinase-9 (MMP-9) levels are elevated in aortic aneurysmal tissue and in that patient plasma, we hypothesized that plasma MMP-9 levels should decrease significantly after conventional and endovascular infrarenal abdominal aortic aneurysm (AAA) repair but that plasma MMP-9 levels would remain elevated in patients with endoleaks. METHODS: A sandwich enzyme-linked immunosorbent assay was used to measure plasma levels of MMP-9 in patients with AAA who underwent conventional (n = 26; mean age, 71.5 years) and endovascular (n = 25; mean age, 76.4 years) AAA repair. Levels were drawn before surgery and at 1 month and 3 months after surgery. Eight patients for endovascular repair had endoleaks identified on postoperative computed axial tomographic scans. RESULTS: No correlation existed between preoperative plasma MMP-9 levels when compared with age, gender, or aneurysm diameter. No significant difference in preoperative plasma MMP-9 levels or AAA diameter was identified between patients with conventional repair compared with endovascular repair. Of the 51 patients, 33 had follow-up samples available for analysis. A significant increase in mean plasma MMP-9 levels was noted 1 month (149.5 +/- 40.1 ng/mL) after conventional AAA repair compared with preoperative levels (83.9 +/- 26.1 ng/mL; P <.05) and remained elevated 3 months after surgery (129.8 +/- 56.6 ng/mL). In those patients who underwent endovascular aneurysm exclusion without endoleak, a significant decrease in mean plasma MMP-9 levels was noted at 3 months (27.4 +/- 5.2 ng/mL) when compared with preoperative values (60.8 +/- 8.8 ng/mL; P <.01). In contrast, patients with endoleak after endovascular exclusion did not have a significant decrease in plasma MMP-9 levels at 3 months. CONCLUSION: Plasma MMP-9 levels remain elevated for as much as 3 months after conventional AAA repair, whereas successful endovascular exclusion of an AAA results in decreased plasma MMP-9 levels by 3 months. MMP-9 may have clinical value as an enzymatic marker for endoleak after endovascular AAA exclusion.     

Parathyroid hormone increases the expression level of matrix metalloproteinase-13 in vivo

Parathyroid hormone (PTH) increases serum calcium (Ca) by enhancing bone resorption and renal Ca reabsorption. However, detailed mechanisms of enhanced bone resorption by PTH remain to be elucidated. Although PTH has been shown to increase the expression level of osteoblastic matrix metalloproteinase (MMP)-13 in vitro, only limited results are available regarding the in vivo regulation of MMP expression. In the present study, we have examined expression levels of MMPs in PTH-infused rats. Infusion of 1.5 or 2.0 nmol/kg/day rat PTH(1–34) for 3 days resulted in a dose-dependent increase in serum Ca. PTH infusion also decreased serum phosphate levels and increased urinary excretion of Ca and phosphate. Infusion of PTH for 7 days resulted in less severe hypercalcemia and hypophosphatemia. Urinary Ca and phosphate excretion in rats infused for 7 days was less than that in rats infused for 3 days. Northern blot analysis showed that PTH infusion increased the expression level of MMP-13 in calvaria, although it did not affect MMP-2 expression. Furthermore, the time-course and severity of hypercalcemia and hypercalciuria correlated with the expression level of MMP-13. In situ hybridization also showed that PTH infusion increased the expression level of MMP-13 in femora. These results indicate that PTH enhances MMP-13 expression in vivo and suggest that PTH stimulates bone resorption at least partly by enhancing MMP-13 expression. Received: June 5, 2000 / Accepted: January 12, 2001     

Activity of matrix metalloproteinase-2 and -9 in abdominal aortic aneurysms. Relation to size and rupture.

OBJECTIVES: to investigate the activity of matrix metalloproteinase (MMP)-2 and -9 in asymptomatic abdominal aortic aneurysms (aAAAs) and ruptured abdominal aortic aneurysms (rAAAs). DESIGN: cross-sectional study. MATERIALS AND METHODS: MMP-2 and MMP-9 activity was estimated in biopsies from the anterior wall of 60 AAAs using gelatin zymography. There were 20 medium-sized (diameter 5<7 cm) aAAAs, 20 large (>57 cm) aAAAs and 20 rAAAs. MMP activity was quantified using a laser densitometer and expressed as arbitrary units (au). RESULTS: mean (SEM) MMP-9 activity was significantly lower in large aAAAs (1190 au +/-247) than in rAAAs (2647 au +/-498, p<0.05). There was no difference in MMP-2 activity. CONCLUSION: High MMP-9 activity in the AAA wall is associated with rupture.     

基质金属蛋白酶-9在腹主动脉瘤体组织中的表达及其临床意义

目的 通过检测基质金属蛋白酶-9（MMP-9）在人腹主动脉瘤（AAAs）组织中的定位表达，探讨MMP-9在腹主动脉瘤发病机制中的作用及临床意义。方法 应用免疫组织化学技术，检测了MMP-9在20例AAAs瘤体组织中的定位表达，并与15例动脉硬化闭塞性疾病（AODs）的病变管壁及10例正常动脉管壁组织中的MMP-9组织学表达进行对照研究。结果 AAAs动脉壁中富含大量MMP-9颗粒，MMP-9阳性表达率达95．0％（19／20）；正常动脉管壁组织未检测到MMP-9的表达；AODs病变血管壁内可见散在分布的MMP-9阳性颗粒，MMP-9阳性表达率为26．7％（4／15）。与AODs和正常动脉管壁相比较，MMP-9在AAAs瘤体组织中的表达差异具有意义（P〈0．01）。结论 腹主动脉瘤体组织中MMP-9的高表达，促进主动脉中层细胞外基质崩解进而导致动脉弹性下降、动脉扩张及进一步的瘤体形成，在AAAs的发病机制中具有重要作用。     

基质金属蛋白酶在腹主动脉瘤形成中的作用研究进展

介绍基质金属蛋白酶在腹主动脉瘤形成中的作用。腹主动脉瘤的形成是多因素共同作用的结果 ,而基质金属蛋白酶对主动脉璧结缔组织的降解 ,导致动脉璧受损 ,是形成动脉瘤的一个重要原因。对于基质金属蛋白酶的深入研究 ,有助于进一步揭示主动脉瘤的发病机制     

Plasma levels of matrix metalloproteinase-9: a possible diagnostic marker of successful endovascular aneurysm repair

ObjectiveThe aim of the study was evaluating the diagnostic value of plasma matrix metalloproteinase- (MMP)-2 and -9 and tissue inhibitor of MMP-1 (TIMP-1) for endoleak detection after endovascular aneurysm repair (EVAR).ReportConsecutive EVAR patients (n = 17) with endoleak and matched controls without endoleak (n = 20) were prospectively enrolled. Increased levels of MMP-9 were observed in patients with endoleak (P < 0.001). Regression analysis showed no significant influence of age, sex or abdominal aortic aneurysm (AAA) size. The receiver operating characteristic (ROC) curve of plasma MMP-9 levels showed that a cut-off value of 55.18 ng ml^?1 resulted in 100% sensitivity and 96% specificity with an AUC value of 0.988 (P < 0.001) to detect endoleak.ConclusionsPlasma MMP-9 levels appear to discriminate between patients with and without an endoleak with high sensitivity and specificity.     

MMP-9在结肠癌组织中表达的临床意义

目的探讨基质金属蛋白酶(MMP-9)在结肠癌组织中的表达的临床意义。方法采用链霉菌抗生物素蛋白-过氧化酶连接(SP)法检测68例结肠癌,26例结肠腺瘤及26例正常结肠组织中MMP-9的表达情况和其与各种临床病理参数的关系,同时分析三者表达的相关性。结果MMP-9在结肠癌组(75%)中的表达明显高于结肠腺瘤组(38.5%)和正常结肠组(15.4%),其在结肠癌组中的表达率也与肌层浸润深度、淋巴转移情况密切相关(P均<0.05)。结论检测MMP-9表达水平对评估结肠癌预后有重要的临床意义,MMP-9的表达可能与结肠癌的浸润、转移有关。     

Nitric oxide modulates expression of matrix metalloproteinase-9 in rat mesangial cells

Nitric oxide modulates expression of matrix metalloproteinase-9 in rat mesangial cells. BACKGROUND: High-output levels of nitric oxide (NO) are produced by rat mesangial cells (MCs) in response to proinflammatory cytokines such as interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) by the inducible isoform of NO synthase (iNOS). We tested modulatory effects of NO on the expression and activities of matrix metalloproteinases-9 and -2 (MMP-9 and MMP-2), respectively. Temporal and spatial expression of these MMPs and their specific inhibitors, the tissue inhibitors of metalloproteinases (TIMPs), seems to be critical in the extensive extracellular matrix (ECM) remodeling that accompanies sclerotic processes of the mesangium. Methods and Results. Using the NO donors S-Nitroso-N-acetyl-D,L-penicillamine (SNAP) and DETA-NONOate, we found strong inhibitory effects of NO mainly on the IL-1beta-induced MMP-9 mRNA levels. NO on its own had only weak effects on the expression of MMP-9 and MMP-2. The addition of the NOS inhibitor NG-monomethyl L-arginine (L-NMMA) dose dependently increased steady-state mRNA levels of cytokine-induced MMP-9, suggesting that endogenously produced NO exerts tonic inhibition of MMP-9 expression. MMP-9 activity in conditioned media from MCs costimulated with IL-1beta and NO donor contained less gelatinolytic activity than media of cells treated with IL-1beta alone. Exogenously added NO did not alter gelatinolytic activity of MMP-9 in cell-free zymographs. The expression levels of TIMP-1 were affected by NO similarly to the expression of MMP-9. CONCLUSION: We conclude that NO modulates cytokine-mediated expression of MMP-9 and TIMP-1 in rat MCs in culture. Our results provide evidence that NO-mediated attenuation of MMP-9 gelatinolytic activity is primarily due to a reduced expression of MMP-9 mRNA, and not the result of direct inhibition of enzymatic activity.     

Increased expression of serum matrix metalloproteinase-9 in patients with moyamoya disease

Background

Moyamoya disease is a chronic occlusive cerebrovascular disease with unknown etiology characterized by an abnormal vascular network at the base of the brain, which can manifest both as ischemic stroke and as cerebral hemorrhage. It was also reported that the patients with moyamoya disease are more vulnerable to cerebral hyperperfusion such as postoperative hemorrhagic complication after extracranial-intracranial bypass surgery despite its low flow revascularization. However, the underlying mechanisms of its pathologic angiogenesis and the occurrence of hemorrhage are undetermined. Excessive degradation of the vascular matrix by MMPs, proteolytic enzymes that degrade all the components of extracellular matrix, can lead to instability of the vascular structure and can thereby cause bleeding. The MMPs also play an important role in tissue remodeling including angiogenesis in both physiologic and pathologic condition.

Methods

We examined the serum levels of MMP-2 and MMP-9 in 16 cases with definitive moyamoya disease by enzyme-linked immunosorbent assay and compared them with those from healthy controls.

Results

The serum MMP-9 level was significantly higher in moyamoya disease (40.18 ng/mL) than in healthy controls (13.75 ng/mL, P = .0372). There was no difference in serum MMP-2 level between moyamoya disease (646.65 ng/mL) and healthy control (677.60 ng/mL). Immunohistochemistry on the surgical specimens showed significant increase in MMP-9 expression within the arachnoid membrane of moyamoya disease.

Conclusion

The increased expression of MMP-9 may contribute to pathologic angiogenesis and/or to the instability of the vascular structure and could thereby cause hemorrhage in moyamoya disease.