高血压患者循环中基质金属蛋白酶-9的研究现状

基质金属蛋白酶(MMPs)是一个家族的锌依赖性蛋白酶,基质金属蛋白酶-9(matrix metalloproteinases-9,MMP-9)又称明胶酶,是金属蛋白酶家族中的一员,它参与细胞外基质(Extracellular matrix,ECM)胶原的代谢,在许多心血管疾病的发生、发展过程中发挥重要作用,如参与心肌梗死后心室的重构和高血压引起的心室重构等.     

Lercanidipine decreases vascular matrix metalloproteinase-2 activity and protects against vascular dysfunction in diabetic rats

This study investigates the participation of the endothelial factors in the α-adrenoceptor contractile responses in mesenteric resistance arteries from 15 days ouabain-treated (25 µg/kg/day) and untreated rats. Ouabain treatment increased blood pressure and heart rate without changing the contractile response to phenylephrine (3 nM–30 µM). Endothelium removal or N^G-nitro-l-arginine methyl ester (l-NAME, 100 µM), increased the responses to phenylephrine. The endothelial modulation was similar in both rat groups, but the l-NAME effects were bigger in arteries from ouabain-treated rats. However, the endothelial NOS expression and the relaxation to acetylcholine (0.1 nM–10 µM) remained unaltered after ouabain treatment. The coincubation with l-NAME and indomethacin (100 µM) leftward shifted the concentration–response curves to phenylephrine in arteries from untreated rats similarly to the displacement after incubation only with l-NAME. However, in mesenteric arteries from treated rats, the co-incubation with indomethacin and l-NAME did not alter the response to phenylephrine. The addition of the inhibitor of calcium activated potassium channels tetraethylammonium (2 mM) further leftward shifted the phenylephrine curves only in arteries from untreated rats. Cyclooxygenase-2 (COX-2) expression was greater in vessels from ouabain-treated rats. In conclusion, the chronic ouabain treatment for 15 days modified the participation of endothelial factors in response to phenylephrine in mesenteric resistance arteries, by increasing the release of NO and prostanoids and impairment the endothelium-derived hyperpolarizing factor (EDHF) release. This was accompanied by an increased COX-2 expression. Although this balance avoids changes in the phenylephrine concentration–response curves, these vascular changes might contribute to maintain the ouabain-induced hypertension.     

The effect of bosentan on matrix metalloproteinase-9 levels in patients with systemic sclerosis-induced pulmonary hypertension

OBJECTIVE: Systemic sclerosis (SSc) is an autoimmune disease that can potentially involve all tissues and organs of the human body. Based on the extent of the disease and organ involvement, different subsets of patients and organ involvement, different subsets of patients have been identified and several classifications proposed have been identified and several classifications proposed aiming to better stratify affected patients. The occurrence aiming to better stratify affected patients. The occurrence of pulmonary arterial hypertension (PAH), characterized of pulmonary arterial hypertension (PAH), characterized by altered tissue remodelling of the entire vessel wall, is the most severe complication that influences prognosis and survival. The molecular basis underlying the vascular damage is not yet known, but a family of enzymes named matrix metalloproteinases (MMPs), with a proteolytic activity towards several extra-cellular matrix (ECM) components, is likely to be involved. Recently, a dual inhibitor of endothelin-1, bosentan, has been successfully evaluated in clinical trials in PAH patients. Research design and method: The aim of this study is to investigate the expression of MMP-2 and MMP-9 in the serum of different subsets of SSc patients, and in patients treated with bosentan. Thirty-five Caucasian patients with SSc were enrolled in the study, 12 of whom were found to have isolated PAH assessed by Doppler echocardiography. Eight patients fully met the inclusion criteria and were eligible for therapy with bosentan given at the dosage of 62.5 mg twice daily for 4 weeks followed by 125 mg twice daily for 50 weeks(15). The remaining patients (4/12) initiated bosentan therapy for a few weeks and, therefore, were considered at the baseline level only. Serum samples were analysed by gelatine zymography. RESULTS: The results suggest that MMP-9 but not MMP-2 is differently expressed according to the degree of organ involvement. In particular, MMP-9 serum levels are significantly decreased in PAH with respect to other subsets of SSc patients. Moreover, in bosentan-treated patients, after 12 months of therapy MMP-9 significantly (p < 0.05) increased and correlated with an improved clinical outcome, as measured by the '6-minutes walking' test. CONCLUSIONS: This is the first time that MMP-9 serum levels are reported to be down-regulated in PAH patients and up-regulated following bosentan treatment. Whether MMP-9 has a pathogenetic role in the vascular damage observed in PAH patients or it is a marker of bosentan effectiveness is not yet known. However, MMP-9 may be an important molecule that needs further investigation in SSc patients to better define its role.     

The effect of bosentan on matrix metalloproteinase-9 levels in patients with systemic sclerosis-induced pulmonary hypertension

Summary

Objective: Systemic sclerosis (SSc) is an autoimmune disease that can potentially involve all tissues and organs of the human body. Based on the extent of the disease and organ involvement, different subsets of patients and organ involvement, different subsets of patients have been identified and several classifications proposed have been identified and several classifications proposed aiming to better stratify affected patients. The occurrence aiming to better stratify affected patients. The occurrence of pulmonary arterial hypertension (PAH), characterized of pulmonary arterial hypertension (PAH), characterized by altered tissue remodelling of the entire vessel wall, is the most severe complication that influences prognosis and survival. The molecular basis underlying the vascular damage is not yet known, but a family of enzymes named matrix metalloproteinases (MMPs), with a proteolytic activity towards several extra-cellular matrix (ECM) components, is likely to be involved. Recently, a dual inhibitor of endothelin-1, bosentan, has been successfully evaluated in clinical trials in PAH patients.

Research design and method: The aim of this study is to investigate the expression of MMP-2 and MMP-9 in the serum of different subsets of SSc patients, and in patients treated with bosentan. Thirty-five Caucasian patients with SSc were enrolled in the study, 12 of whom were found to have isolated PAH assessed by Doppler echocardiography. Eight patients fully met the inclusion criteria and were eligible for therapy with bosentan given at the dosage of 62.5 mg twice daily for 4 weeks followed by 125 mg twice daily for 50 weeks¹⁵. The remaining patients (4/12) initiated bosentan therapy for a few weeks and, therefore, were considered at the baseline level only. Serum samples were analysed by gelatine zymography.

Results: The results suggest that MMP-9 but not MMP-2 is differently expressed according to the degree of organ involvement. In particular, MMP-9 serum levels are significantly decreased in PAH with respect to other subsets of SSc patients. Moreover, in bosentan-treated patients, after 12 months of therapy MMP-9 significantly (p < 0.05) increased and correlated with an improved clinical outcome, as measured by the '6-minutes walking' test.

Conclusions: This is the first time that MMP-9 serum levels are reported to be down-regulated in PAH patients and up-regulated following bosentan treatment. Whether MMP-9 has a pathogenetic role in the vascular damage observed in PAH patients or it is a marker of bosentan effectiveness is not yet known. However, MMP-9 may be an important molecule that needs further investigation in SSc patients to better define its role.     

Anacardic Acid inhibits the catalytic activity of matrix metalloproteinase-2 and matrix metalloproteinase-9

Cashew nut shell liquid (CNSL) has been used in traditional medicine for the treatment of a wide variety of pathophysiological conditions. To further define the mechanism of CNSL action, we investigated the effect of cashew nut shell extract (CNSE) on two matrix metalloproteinases, MMP-2/gelatinase A and MMP-9/gelatinase B, which are known to have critical roles in several disease states. We observed that the major constituent of CNSE, anacardic acid, markedly inhibited the gelatinase activity of 3T3-L1 cells. Our gelatin zymography studies on these two secreted gelatinases, present in the conditioned media from 3T3-L1 cells, established that anacardic acid directly inhibited the catalytic activities of both MMP-2 and MMP-9. Our docking studies suggested that anacardic acid binds into the MMP-2/9 active site, with the carboxylate group of anacardic acid chelating the catalytic zinc ion and forming a hydrogen bond to a key catalytic glutamate side chain and the C15 aliphatic group being accommodated within the relatively large S1' pocket of these gelatinases. In agreement with the docking results, our fluorescence-based studies on the recombinant MMP-2 catalytic core domain demonstrated that anacardic acid directly inhibits substrate peptide cleavage in a dose-dependent manner, with an IC(50) of 11.11 μM. In addition, our gelatinase zymography and fluorescence data confirmed that the cardol-cardanol mixture, salicylic acid, and aspirin, all of which lack key functional groups present in anacardic acid, are much weaker MMP-2/MMP-9 inhibitors. Our results provide the first evidence for inhibition of gelatinase catalytic activity by anacardic acid, providing a novel template for drug discovery and a molecular mechanism potentially involved in CNSL therapeutic action.     

Inhibitory profiles of captopril on matrix metalloproteinase-9 activity

To investigate the inhibitory profiles of angiotensin-converting enzyme (ACE) inhibitors on matrix metalloproteinase-9 (MMP-9) activity, the inhibitory activity and molecular interaction of captopril on human MMP-9 were studied. Plasma MMP-9 and ACE activities in samples from patient with acute myocardial infarction were similarly inhibited by captopril. Molecular models showed that captopril directly bound to the MMP-9 active center. Compared with the other ACE inhibitors, the compact structure of captopril seemed to make the inhibitory profiles on the MMP-9 active site.     

阻塞性睡眠呼吸暂停低通气综合征患者血清基质金属蛋白酶-9水平的变化

目的 观察阻塞性睡眠呼吸暂停低通气综合征(OSAHS)患者血清基质金属蛋白酶-9(MMP-9)水平的变化。方法 用酶联免疫吸附法对20例OSAHS患者进行血清MMP-9水平进行测定,与16例健康者(对照组)进行比较。结果 OSAHS患者血清MMP-19水平较正常对照组升高,有统计学意义。MMP-9升高与阻塞性睡眠呼吸暂停的严重程度相关。结论 OSAHS患者存在MMP-9代谢的异常,血清MMP-9可反映OSAHS的病情严重程度。     

颈动脉粥样硬化斑块患者血清基质金属蛋白酶9的表达及意义

目的 探讨颈动脉粥样硬化斑块与血清基质金属蛋白酶9 (MMP-9)水平的关系.方法 选取30例经彩色多普勒超声证实存在颈动脉粥样硬化斑块的患者为研究组,其中稳定斑块18例,不稳定斑块12例,30例健康查体者为对照组.均抽取空腹血应用酶联免疫吸附法检测血清MMP-9水平.结果 研究组的MMP-9表达为(295.34±126.76) μg/L,明显高于对照组的(98.38±34.57)μg/L(t =4.835,P＜0.05);研究组稳定斑块患者中MMP-9的表达为(267.45±113.86) μg/L,明显低于不稳定斑块患者的(498.57±142.68) μg/L(t =3.768,P＜0.05).结论 血清MMP-9水平与颈动脉粥样硬化斑块形成有关,可以反映颈动脉粥样硬化斑块的稳定性.     

冠状动脉粥样硬化性心脏病患者血清基质金属蛋白酶9检测中的质量控制

目的 通过检测不同类型冠状动脉粥样硬化性心脏病（冠心病）患者血清基质金属蛋白酶9（MMP-9）和质控样本,探讨＂即刻性＂质控法在科研实验室酶联免疫吸附法（ELISA）中的应用价值.方法 采用ELISA检测冠心病患者血清MMP-9,用＂即刻性＂质控法进行室内质量控制.结果 ELISA检测MMP-9板内变异系数值≤15%,血清MMP-9质控测定结果SI上限值为1.15,SI下限值为0.625,经与SI值表中第3天的2倍标准差进行比较,均在允许范围之内;与健康对照组、心绞痛患者组相比较,心肌梗死组患者血清MMP-9水平明显升高（P＜0.05）.结论 ＂即刻性＂质控法对科研实验室是一种行之有效的室内质量控制方法.     

Lercanidipine in type II diabetic patients with mild to moderate arterial hypertension

This study evaluates the effects of lercanidipine antihypertensive treatment on glucose homeostasis in patients with type II diabetes mellitus with mild to moderate hypertension. Forty patients were enrolled. After a 2-week wash-out period, they were randomly allocated to receive in double-blind manner either 10 mg or 20 mg in single daily administration for 8 weeks. Nonresponding patients after the initial 4 weeks, were titrated up to 20 mg and 30 mg lercanidipine, respectively. At the end of the double-blind treatment, all patients entered in single-blind 4 weeks placebo follow-up. Systolic and diastolic blood pressure significantly decreased in both groups of patients after 4 weeks of treatment, and decreased further during the following 4 weeks. In both groups, progressive and significant decrease in fasting blood glucose, glycosylated hemoglobin and area under the curve of the oral glucose tolerance test were detected during lercanidipine treatment. Similarly, a decrease in serum fructosamine values were also observed. All variables returned to towards baseline values during the placebo follow-up period. Adverse events (headache and mild asthenia) were limited to two patients and resolved spontaneously. These data indicate that lercanidipine is effective in lowering high blood pressure in hypertensive patients with type II diabetes mellitus and does not exert negative effects on glucose homeostasis.     

辛伐他汀对急性冠脉综合征患者血清基质金属蛋白酶-9及其组织抑制因子-1的影响

目的观察辛伐他汀对急性冠脉综合征（ACS）患者血清基质金属蛋白酶-9（MMP-9）及其组织抑制因子-1（TIMP-1）的影响。方法选择ACS患者4JD例（ACS组）、稳定性心绞痛（SAP）患者34例（SAP组）、健康体检者25例（对照组）,采用酶联免疫吸附法测定其血清可溶性TIMP-1及MMP-9水平,辛伐他汀治疗后复查TIMP-1及MMP-9水平变化。结果ACS组血清MMP-9水平（930±127）μg/L高于SAP组的（619±114）μg／L及对照组的（417±105）μg／L（均P〈0．05）,而TIMP-1水平（73±14）μg／L显著低于SAP组的（121±17）μg/L及对照组的（151±24）μg／L（均P〈0．05）;辛伐他汀治疗4周后ACS组血清MMPO水平为（641±123）μg／L、TIMP-1水平为（114±18）μg/L,与治疗前比较差异均有统计学意义（P〈0．01,P〈0．05）,SAP组治疗后MMP-9水平为（546±122）μg/L,与治疗前比较差异具有统计学意义（P〈0．05）,而TIMP-1水平无明显改变（P〉0．05）。结论MMP-9及TIMP-1可能与ACS发病密切相关,监测其血清水平对ACS的治疗有重要的指导意义。     

Inhibition of matrix metalloproteinase-9 activity by lisinopril after myocardial infarction in hamsters

We measured angiotensin-converting enzyme and matrix metalloproteinase-9 activities after myocardial infarction in hamsters and compared the effects of an angiotensin-converting enzyme inhibitor lisinopril with those of an angiotensin receptor blocker candesartan cilexetil after myocardial infarction. Angiotensin-converting enzyme activity was significantly increased 3 and 7 days, but not 1 day after myocardial infarction. Matrix metalloproteinase-9 activity was significantly increased 1 day after myocardial infarction. Lisinopril significantly inhibited both angiotensin-converting enzyme and matrix metalloproteinase-9 activities, but candesartan cilexetil did not. Angiotensin-converting enzyme inhibitors might directly inhibit matrix metalloproteinase-9 activity.     

基质金属蛋白酶9水平在脑梗死出血转化中的临床意义

目的 探讨血浆基质金属蛋白酶9(MMP-9)水平在急性脑梗死出血转化中的临床意义.方法 157例发病24 h内急性脑梗死患者,根据是否存在出血转化分为脑梗死组(梗死组,135例)和脑梗死出血转化组(转化组,22例);于起病24 h急诊行血浆MMP-9水平测定,并分析其与脑梗死出血转化、梗死体积大小和病情严重程度的相关性.结果 转化组血浆MMP-9水平高于梗死组(P＜0.01);梗死体积大者血浆MMP-9水平也明显高于中、小梗死者(P＜0.01);NIH卒中量表(NIHSS)评分≥7分者血浆MMP-9的含量亦明显上调(P＜0.01).结论 血浆MMP-9水平与脑梗死出血转化的发生及梗死体积大小、病情严重程度相关.脑梗死早期检测血浆MMP-9可作为临床预测脑梗死出血转化的参考指标.     

Expression of matrix metalloproteinase-9 in thyroid tumors tissue

目的 探讨基质金属蛋白酶-9(MMP-9)在甲状腺肿瘤组织的表达情况.方法 采用免疫组织化学SP法检测20例瘤旁甲状腺组织、20例甲状腺腺瘤和53例甲状腺癌的石蜡标本MMP-9表达水平.结果 MMP-9在甲状腺腺癌的表达率显著高于甲状腺腺瘤(96.41% vs 55%,P＜0.01),瘤旁甲状腺组织未见表达.结论 甲状腺肿瘤存在MMP-9的过表达.     

基质金属蛋白酶-9在动脉粥样硬化血管中的异常表达

目的了解基质金属蛋白酶-9在Apo-E基因敲除小鼠的动脉粥样硬化斑块血管中的表达。方法 8只5周龄,雄性SPF级C57BL/6J小鼠为对照组(NS组)。8只相同周龄,雄性SPF级载脂蛋白E基因敲除(C57BL/6J-ApoE-/-)小鼠为实验组(FS组)。高脂(1%胆固醇+15%猪油)喂食15周,20周龄取材检测。结果 FS组和NS组体重分别为(29.10±2.53)g和(33.20±5.18)g,差异无统计学有意义(P>0.05)。FS组较NS组血清总胆固醇显著增高,分别为(13.52±1.58)mEq/L和(2.12±0.35)mEq/L,差异有统计学有意义(P<0.01)。NS组主动脉管壁正常,无AS斑块;FS组主动脉出现典型的AS斑块,胶原纤维明显减少。免疫组化显示NS组可见主动脉血管内、中膜MMP-9呈散在弱阳性表达;而其在FS组则是内膜表达明显增高,呈强阳性表达。FS组主动脉弓MMP-9在mRNA及蛋白水平的表达较NS组均明显增高。mRNA平均光密度值为(11.61±1.92)和(1.52±0.40),差异有统计学意义(P<0.05);蛋白平均光密度值为(190.40±102.71)和(40.19±26.83),差异有统计学有意义(P<0.01)。结论在高脂喂食的载脂蛋白E基因敲除小鼠动脉粥样硬化模型中,MMP-9高表达可能与动脉粥样硬化斑块的形成和胶原纤维明显减少有关。     

氨基葡萄糖对EAE大鼠BBB和MMP-9的作用(英文)

本研究旨在探讨氨基葡萄糖(GS)对实验性自身免疫性脑脊髓炎(EAE)大鼠血脑屏障(BBB)功能及基质金属蛋白酶-9(MMP-9)的影响。雌性Wistar大鼠随机分为EAE组、GS组和佐剂组。EAE组和GS组用完全抗原和减毒百日咳原液免疫后腹腔分别注射4.5 mL/(kg·d)的磷酸盐缓冲液(PBS)和180 mg/(kg·d)的GS;佐剂组用完全弗氏佐剂和减毒百日咳原液免疫后腹腔注射4.5 mL/(kg·d)自PBS;免疫后(p.i.)6、8、10、12、14、16、18天检测大鼠血清和脑脊液(CSF)中白蛋白(ALB)含量,其CSF与血清ALB比值(QA)作为评价BBB功能及动态变化的指标。同时取大鼠脑和脊髓制成石蜡切片,行MMP-9免疫组化染色。结果显示,EAE组QA值和MMP-9在免疫后初期开始升高,p.i.10天达高峰,二者相关性分析显示呈显著正相关,并且其大小同临床症状的严重程度相关联。GS能明显改善EAE临床症状(P<0.05),降低EAE发病时QA值水平和中枢神经系统内MMP-9的表达(P<0.05)。我们认为GS可抑制MMP-9的生成,减轻BBB的破坏,对EAE发病起一定程度的干预作用。     

基质金属蛋白酶-9在甲状腺肿瘤组织中的表达

目的探讨基质金属蛋白酶-9(MMP-9)在甲状腺肿瘤组织的表达情况。方法采用免疫组织化学SP法检测20例瘤旁甲状腺组织、20例甲状腺腺瘤和53例甲状腺癌的石蜡标本MMP-9表达水平。结果 MMP-9在甲状腺腺癌的表达率显著高于甲状腺腺瘤(96.41%vs 55%,P<0.01),瘤旁甲状腺组织未见表达。结论甲状腺肿瘤存在MMP-9的过表达。     

基质金属蛋白酶-9与糖尿病肾病的相关性

目的观察大鼠糖尿病肾病(DN)发展过程中肾脏局部基质金属蛋白酶-9(MMP-9)表达的改变,分析其与DN的相关性。方法对16只链脲佐菌素(STZ)糖尿病大鼠模型分别于造模后第4、8周通过免疫组织化学染色法观察肾脏局部MMP-9的表达,以MMP-9平均积分吸光度(IDP)为观察指标。结果正常对照组第4、8周大鼠肾脏MMP-9表达(IDP分别为15.6±73.4,16.2±3.1)呈强阳性;实验组第4、8周大鼠肾脏MMP-9的表达(IDP分别为8.2±0.9,3.9±0.6)呈递减(P<0.01)。结论MMP-9表达减少是DN早期病理改变的重要原因之一。     

脂多糖对人类风湿性关节炎滑膜细胞基质金属蛋白酶-9表达的影响

目的 研究脂多糖（LPS）对人类风湿性关节炎（RA）成纤维状滑膜细胞(FLS)基质金属蛋白酶-9（MMP-9）表达的影响。方法 明胶酶谱法测定MMP-9酶活性；Western blot法测定MMP-9蛋白的表达；RT-PCR法测定MMP-9 mRNA的表达。结果 LPS处理对FLS中MMP-9表达无显著影响；LPS刺激的U937细胞培养上清液可明显增强FLS 中MMP-9酶活性、蛋白分泌及mRNA表达；地塞米松可显著抑制上述变化，且其抑制作用随浓度的增加而增强。结论 LPS对FLS中MMP-9表达无直接影响，LPS刺激的U937细胞上清液使FLS中MMP-9表达增加，而地塞米松能抑制MMP-9的变化。     

高压氧治疗对脑梗死患者血清基质金属蛋白酶-9的影响

目的探讨高压氧（HBO）治疗后急性脑梗死患者血清基质金属蛋白酶-9（MMP-9）水平的动态变化及其与临床疗效的关系。方法采用酶联免疫吸附双抗体夹心法测定脑梗死患者入院时、发病后第3、5、10天以及正常对照组血清MMP-9浓度。对所有患者于入院时和治疗后10d、1个月进行神经功能缺损评分,评价HBO组和常规组疗效差异。结果与正常对照组比较,HBO组和常规组入院时血清MMP-9的浓度均明显增高（P〈0.01）。与治疗前比较,常规治疗组和HBO治疗组血清MMP-9水平均呈现先增高后降低的动态变化趋势,3d时达高峰,于治疗后10d时降低至入院时的水平,但仍高于正常对照组（P〈0.01）,比较2组患者各时间点的MMP-9的浓度显示,在治疗后3、5d时HBO治疗组的血清MMP-9浓度低于同期常规治疗组（P〈0.05）;治疗1个月后神经功能缺失评分HBO组明显低于常规组;HBO治疗组疗效优于常规治疗组（P〈0.05）;脑梗死患者神经功能缺损评分减少值与患者发病后第3天血清MMP-9水平呈负相关（r=-0.625,P〈0.01）。结论 HBO治疗能降低脑梗死患者升高的血清MMP-9浓度,减少神经功能缺失程度,其临床疗效可能与降低患者血清MMP-9的浓度有关。