血浆中性粒细胞弹性蛋白酶与冠心病的关系

目的 :探讨血浆中性粒细胞弹性蛋白酶 (neutrophilelastase,NE)与冠心病 (CHD)的关系。方法 :对85例观察对象进行分组 ,根据冠状动脉造影结果分为 :简单病变组 2 4例 ,复杂病变组 5 1例 ,正常对照组 10例。根据CHD病情分为 :急性心肌梗死 (AMI)组 9例 ,不稳定型心绞痛 (UAP)组 4 5例 ,稳定型心绞痛 (SAP)组 2 1例 ,正常对照组 10例。用酶联免疫吸附法 (ELASA)检测血浆NE含量 ,比较不同组间NE含量。结果 :血浆NE含量 :复杂病变组 [(6 3.4 2± 13.6 2 ) μg/L]高于简单病变组 [(4 4 .5 0± 12 .73) μg/L],差异有统计学意义 (P <0 .0 1)。AMI组 [(71.4 8± 10 .16 ) μg/L]高于UAP组 [(6 1.2 7± 14 .5 7) μg/L],但差异无统计学意义 (P >0 .0 5 ) ,明显高于SAP组 [(4 0 .95± 10 .0 9) μg/L]及正常对照组 [(36 .6 3± 12 .35 ) μg/L],差异均有统计学意义 (P <0 .0 1) ,后两组间比较差异无统计学意义 (P >0 .0 5 )。UAP组明显高于SAP组及正常对照组 ,差异有统计学意义 (P <0 .0 1)。结论 :血浆NE含量与冠状动脉病变稳定性及CHD病情严重性相关 ,NE可能是冠状动脉病变活动性的标志物。     

金属蛋白酶抑制剂对大鼠结肠炎的作用研究

炎症性肠病（IBD）包括溃疡性结肠炎（UC）和克罗恩病（CD）,为非特异性肠道炎症,其病程迁延,治疗困难.最近研究提示,IBD炎症组织中基质金属蛋白酶（MMP）表达增高,且MMP抑制剂（MMPI）治疗一些胃肠道疾病有一定疗效[1],但对慢性肠道黏膜炎症的治疗尚鲜见报道.本研究拟采用三硝基苯磺酸（TNBS）二次致炎法,建立急、慢性结肠炎共存的动物模型,检测MMPI--二氮杂菲防治TNBS肠炎的作用,并与传统的柳氮磺胺吡啶（SASP）比较,希望能为IBD的治疗提供新的思路和途径.     

双歧杆菌对小鼠急性实验性结肠炎作用的研究

目的 研究双歧杆菌在葡聚糖硫酸钠(DSS)诱导的急性溃疡性结肠炎小鼠模型中的作用.方法 将80只BALB/C小鼠均分为8组,每组10只.除正常对照组、单纯0501菌株组、单纯c122菌株组外,其他5组动物均给予5%DSS造模7 d.在造模开始前2 d,阴性对照组用0.9%氯化钠液灌肠、阳性对照组用柳氮磺胺吡啶(SASP)20 mg/ml灌肠、DSS+0501菌株组用1×109 CFU/ml 0501菌液灌肠、DSS+c122菌株组用1×109 CFU/ml c122液菌灌肠.9 d后处死动物取结肠标本,行H-E染色,观察镜下结肠病理变化,并用免疫组化染色、RT-PCR技术分析结肠黏膜中白细胞介素(IL)-10 mRNA及蛋白表达.结果 DSS造模过程中给予双歧杆菌灌肠小鼠(0501菌株组和c122菌株组)的结肠炎性反应程度较模型组加重,结肠黏膜IL-10表达减少.而单纯给予双歧杆菌灌肠小鼠(单纯0501菌株组和单纯c122菌株组)未见结肠炎表现.结论 双歧杆菌的某些菌株在结肠黏膜屏障功能受损情况下,可加重溃疡性结肠炎小鼠的结肠黏膜损伤.     

中性粒细胞弹性蛋白酶及其抑制剂与肺癌关系的研究进展

肿瘤转移作为一系列复杂事件的结果,这一过程需要很多酶的参与.中性粒细胞弹性蛋白酶在肺癌侵袭和转移中发挥重要作用,在生理条件下,中性粒细胞弹性蛋白酶有很多特殊的作用底物,过量的中性粒细胞弹性蛋白酶可导致弹力蛋白的降解,还可导致细胞外基质的降解.肺癌组织中的中性粒细胞弹性蛋白酶的量不仅可作为肺癌预后的独立指标,而且在重度联合免疫缺陷小鼠的肺癌种植模型中,一种特殊的中性粒细胞弹性蛋白酶抑制剂ONO-5046完全抑制了肺癌细胞的生长.中性粒细胞弹性蛋白酶免疫抑制剂的应用有望成为阻止肺癌侵袭和转移的有效方法.     

中性粒细胞性弹性蛋白酶与炎症性肺部疾病

中性粒细胞弹性蛋白酶(NE)是由多形核中性粒细胞释放的弹性蛋白酶,在肺部炎症性疾病发病机制中,既有有利作用,又有不利作用,与体内NE-抗NE平衡有关。本文又综述了通过维持NE-抗NE平衡来治疗几种炎症性疾病的研究进展及目前存在的问题。     

中性粒细胞弹性蛋白酶抑制剂对颅脑损伤大鼠脑组织的保护作用

目的探讨中性粒细胞弹性蛋白酶(NE)抑制剂西维来司钠对颅脑损伤(TBI)大鼠脑组织的保护作用及机制。方法 SD大鼠70只,随机分为对照组(n=10)、TBI组(n=30)和干预组(n=30),TBI组和干预组又分为24、72、168 h亚组。用液压打击方法制备TBI模型,TBI组术后立即腹腔注射生理盐水,干预组术后立即腹腔注射西维来司钠。于术后24、72和168 h取挫伤侧脑组织标本,测定NE、超氧化物歧化酶(SOD)、丙二醛(MDA)和髓过氧化物酶(MPO)活性和含量。结果与对照组相比,TBI组脑组织NE、MDA含量和MPO活性明显增高(P<0.05),SOD活性明显降低(P<0.05);与TBI组相比,干预组脑组织NE、MDA含量和MPO活性明显降低(P<0.05),SOD活性明显增高(P<0.05)。结论西维来司钠可以通过抑制NE活性,抑制中性粒细胞活化和浸润,减少自由基的产生及脂质过氧化,保护神经细胞和内皮细胞的膜性结构和功能的完整,对TBI大鼠脑组织具有保护作用。     

中性粒细胞弹性蛋白酶与慢性阻塞性肺疾病

中性粒细胞弹性蛋白酶是丝氨酸蛋白酶超家族成员之一,它能够消化、降解细胞外基质和上皮连接结构造成气道上皮完整性破坏和肺组织损伤,并通过促进炎性细胞聚集,引起黏液高分泌等机制促进慢性阻塞性肺疾病气道炎症与重构的发生。     

中性粒细胞性弹性蛋白酶与炎症性肺部疾病

中性粒细胞弹性蛋白酶（NE）是由多形核中性粒细胞释放的弹性蛋白酶，在肺部炎症性疾病发病机制中，既有有利作用，又有不利作用，与体内NE－抗NE平衡有关。本文又综述了通过维持NE－抗NE平衡来治疗几种炎症性疾病的研究进展及目前存在的问题。     

中性粒细胞弹性蛋白酶在急性肺损伤发病中的作用

急性肺损伤的发病机制尚不清楚，但中性粒细胞在肺部聚集并释弹性蛋白酶具有重要作用。中性粒细胞弹性蛋白酶能分解内皮细胞外基质，增加血管通透性，促进血浆蛋白和活性物质的外漏，引起非心源性肺水肿，为对抗其活性，可采取抗蛋白酶，抗氧化剂以及表面活性物质替代治疗等措施。     

川芎嗪对实验性溃疡性结肠炎治疗作用的研究

目的：研究川芎嗪对小鼠溃疡性的炎的治疗作用及其机制。方法：在小鼠溃疡性结肠炎模型上观察川芎嗪的治疗作用,并用免疫组化LSAB（labelled streptavidin biotin)法检测肠组织中P选择素的表达,用ELISA法检测血浆P选择素含量。结果：溃疡性结肠炎小鼠结肠粘膜充血、水肿、糜烂或溃疡性形成;粘膜炎症细胞浸润（主要为中性粒细胞）及隐窝脓肿形成;川芎嗪治疗组小鼠未见明显病理变化,疾病活动指数（DAI）和病理学评分明显改善。免疫组化和ELISA检测发现,溃疡性结肠炎时肠组织P选择素表达和血中P选择素水平显著升高,经川芎嗪治疗的小鼠P选择素明显下降。结论：P选择素与溃疡性结肠炎密切相关,川芎嗪对溃疡性结肠炎具有治疗作用。     

Inhibition of neutrophil elastase prevents the development of murine dextran sulfate sodium-induced colitis

Background Neutrophil elastase (NE) is a major secretory product from activated neutrophils and a major contributor to tissue destruction. However, little is known about the pathogenic contribution of NE to ulcerative colitis (UC). This study was designed to investigate the contribution of NE by measuring NE activity in plasma and colonic mucosal tissue from UC patients and a murine acute colitis model, and to elucidate the therapeutic effect of the NE-specific inhibitor ONO-5046. Methods The NE enzyme activities in plasma and colonic mucosal tissue from UC patients were directly measured using an enzyme–substrate reaction. Acute colitis was induced in mice by administration of 1.5% dextran sulfate sodium (DSS) for 5 days. DSS-induced colitis mice were then treated with ONO-5046 (50 mg/kg body weight) intraperitoneally twice a day. Results In UC patients, the NE enzyme activity was significantly elevated in both the plasma and colonic mucosal tissue compared with healthy controls. In DSS-induced colitis mice, the NE enzyme activity increased in parallel with the disease development. ONO-5046 showed therapeutic effects in DSS-treated mice by significantly reducing weight loss and histological score. ONO-5046 suppressed the NE enzyme activities in both plasma and culture supernatant of colonic mucosa from DSS-induced colitis mice. Conclusions ONO-5046, a specific NE inhibitor, prevented the development of DSS-induced colitis in mice. NE therefore represents a promising target for the treatment of UC patients.     

基因芯片技术分析川芎嗪对小鼠实验性结肠炎的影响

目的 用基因芯片技术研究川芎嗪对实验性结肠炎小鼠基因表达谱的影响.方法 健康昆明小鼠30只,均分为对照组、0.9%氯化钠组及川芎嗪组.除对照组外,其余小鼠均以恶唑酮灌肠造模.提取0.9%氯化钠组及川芎嗪组结肠组织mRNA,制备cDNA探针,分别用Cy3和Cy5两种荧光染料标记,与基因表达谱芯片进行杂交,经扫描、分析,得出药物作用后表达有差异的基因,并应用荧光定量RT-PCR技术对其中2个表达差异明显的基因(白细胞介素-10和白细胞介素-4)进行验证.结果 川芎嗪组差异表达基因432个,占芯片基因总数的2.86%.其中表达上调的基因307个,表达下调的基因125个,其中部分基因已知其功能.结论 川芎嗪通过抑制肠道炎性反应、抗粘附分子及改善免疫功能等多种作用途径为治疗溃疡性结肠炎提供了理论上的可能,但更为精细的治疗靶点还有待进一步探索.     

c-src/NADPH氧化酶1/活性氧对中性粒细胞弹力蛋白酶诱导气道黏蛋白5 AC表达的影响

目的 探讨中性粒细胞弹力蛋白酶(NE)诱导气道黏液高分泌的上游信号调节机制.方法 体外培养人气道BEAS-2B上皮细胞,将细胞分为空白对照组(不加任何刺激)、NE组(NE刺激)、NE+PP2组(NE刺激合并c-src抑制剂PP2)、NE+c-src siRNA组(NE刺激合并c-src siRNA)、NE+Noxl siRNA组[NE刺激合并NADPH氧化酶1(Nox1)siRNA]、阴件siRNA对照组(加入阴性对照siRNA)及NE+阴性siRNA组(NE刺激合并阴性对照siRNA)为干预条件.检测干预前后各组细胞巾活性氧含量、NoxI蛋白含量、黏蛋白5AC的蛋白及mRNA水平.用四甲基偶氮唑盐法测定各组细胞活性;活性氧试剂盒测定活性氧相对含量;逆转录PCR法检测各组黏蛋白5AC mRNA水平;ELISA法分析细胞黏蛋白5AC的蛋白相对含量;Western blot法检测Noxl蛋白及c-src蛋白的相对含量.结果 NE组细胞中Nox1蛋白相对含量为0.88±0.12,高于空白对照组的0.32±0.09(t=9.12.P=0.003);活性氧相对含量为0.76±0.09,高于空白对照组的0.18±0.02(t=9.44,P=0.003);黏蛋白5AC蛋白相对含量为0.82±0.09,基因转录水平为0.77±0.05,均高于空白对照组(分别为0.21±0.11和0.18±0.08,t值分别为7.75和6.13,P值分别为0.004和0.006).NE+c-src siRNA组细胞的Nox1蛋白相对含最(0.39±0.08)、活性氧相对含量(0.29±0.05)均低于NE组(t值分别为5.43和5.60,均P=0.007);黏蛋白5AC蛋白相对含量(0.38±0.09)及mRNA水平(0.41±0.04)低于NE组(t值分别为5.28和4.09,P值分别为0.008和0.034).NE+PP2组活性氧相对含量为0.41±0.11,Nox1蛋白相对含量为0.44±0.05,黏蛋白5AC蛋白及mRNA水平分别为0.48±0.08和0.46±0.07,均低于NE组(均P<0.05).NE+Noxl siRNA组中活性氧相对含量(0.19±0.06)、黏蛋白5AC蛋白相对含量(0.31±0.05)及mRNA水平(0.32±0.06)也低于NE组(均P<0.05).结论 c-src/Noxl参与了NE诱导的活性氧活化,是气道上皮细胞黏蛋白5AC合成及分泌的上游信号调节因子.     

Effect of a neutrophil elastase inhibitor on ventilator-induced lung injury in rats

Objective

We hypothesized that pretreatment with sivelestat therapy could attenuate ventilator-induced lung injury (VILI) and lung inflammation in a rat model.

Methods

The neutrophil elastase inhibitor was administered intraperitoneally 30 min before and at the initiation of ventilation. The rats were categorized as (I) sham group; (II) VILI group; (III) sivelestat group; (IV) early sivelestat group. Wet-to-dry weight ratio, bronchoalveolar lavage fluid (BALF) neutrophil and protein, tissue malondialdehyde (MDA) and histologic VILI scores were investigated.

Results

The ratio of wet-to-dry weight, BALF neutrophil and protein, tissue MDA and VILI scores were significantly increased in the VILI group compared to the sham group [3.85±0.32 vs. 9.05±1.02, P<0.001; (0.89±0.93)×10⁴ vs. (7.67±1.41)×10⁴ cells/mL, P<0.001; 2.34±0.47 vs. 23.01±3.96 mg/mL, P<0.001; 14.43±1.01 vs. 36.56±5.45 nmol/mg protein, P<0.001; 3.78±0.67 vs. 7.00±1.41, P<0.001]. This increase was attenuated in the early sivelestat group compared with the sivelestat group [wet-to-dry ratio: 6.76±2.01 vs. 7.39±0.32, P=0.032; BALF neutrophil: (5.56±1.13)×10⁴ vs. (3.89±1.05)×10⁴ cells/mL, P=0.021; BALF protein: 15.57±2.32 vs. 18.38±2.00 mg/mL, P=0.024; tissue MDA: 29.16±3.01 vs. 26.31±2.58, P=0.049; VILI scores: 6.33±1.41 vs. 5.00±0.50, P=0.024].

Conclusions

Pretreatment with a neutrophil elastase inhibitor attenuates VILI in a rat model.     

葡聚糖硫酸钠致大鼠急性溃疡性结肠炎模型建立与评价

目的探讨葡聚糖硫酸钠（dextran sulfate sodium,DSS）诱导大鼠溃疡性结肠炎（ulcerative colitis,UC）急性期模型的建立。 方法60只SD大鼠,雌雄各半,随机分成空白组（n=12）、模型组（n=48）,空白组给予自由饮食,模型组给予3%DSS溶液自由饮用7天。应用疾病活动指数（DAI）及组织学评分（HI）对各组进行对比。 结果模型组DAI及HI评分与空白组比较均有统计学意义（P<0.01）。模型组均不同程度的出现腹泻、隐血试验阳性、脓血便,病理可见炎症主要累及黏膜和黏膜下层,上皮内杯状细胞减少,隐窝破坏。 结论DSS诱导的UC急性期模型的临床和病理表现与人类UC相似,是理想的UC模型之一。     

Expression of neutrophil elastase and myeloperoxidase mRNA in patients with newly diagnosed type 2 diabetes mellitus

Background and aimsNeutrophil elastase and myeloperoxidase enzymes protect us from infection by killing pathogens. However, exaggerated activities of these enzymes can induce tissue damage, inflammation and oxidative stress. The present study was aimed to explore the expressions of neutrophil elastase and myeloperoxidase mRNA in the peripheral blood leukocytes (PBL) in patients with newly diagnosed type 2 diabetes mellitus.MethodsIn this cross-sectional study, 104 participants including 65 normoglycemic control subjects and 39 newly diagnosed type 2 diabetes patients were recruited. Glycemic and metabolic markers were evaluated from fasting blood samples. The mRNA levels of neutrophil elastase and myeloperoxidase genes in the PBL were quantified by real-time quantitative PCR.ResultsCompared to control subjects, diabetes patients showed a significant down regulation of both neutrophil elastase (p = 0.039) and myeloperoxidase (p = 0.023) mRNA expressions in the PBL. The neutrophil elastase and myeloperoxidase mRNA levels showed a negative trend with fasting glucose levels but did not show any significant correlations with HbA1c, insulin level, insulin resistance or sensitivity status.ConclusionsIt was concluded that type 2 diabetes mellitus is associated with a decrease in neutrophil elastase and myeloperoxidase gene expression in the PBL.     

Effects of neutrophil elastase inhibitor in patients undergoing esophagectomy:A systematic review and meta-analysis

AIM: To evaluate the benefit and safety of sivelestat(a neutrophil elastase inhibitor) administration in patients undergoing esophagectomy. METHODS: Online databases including Pub Med, EMBASE, the Cochrane Library, Web of Knowledge, and Chinese databases(Wanfang database, VIP and CNKI) were searched systematically up to November 2013. Randomized controlled trials and high-qualitycomparative studies were considered eligible for inclusion. Three reviewers evaluated the methodological quality of the included studies, and Stata 12.0 software was used to analyze the extracted data. The risk ratio(RR) was used to express the effect size of dichotomous outcomes, and mean difference(MD) or standardized mean difference was used to express the effect size of continuous outcomes.RESULTS: Thirteen studies were included in this systematic review and nine studies were included in the meta-analysis. The duration of mechanical ventilation was significantly decreased in the sivelestat group on postoperative day 5 [I2 = 76.3%, SMD =-1.41, 95%CI:-2.63-(-0.19)]. Sivelestat greatly lowered the incidence of acute lung injury in patients after surgery(I2 = 0%, RR = 0.27, 95%CI: 0.08-0.93). However, it did not decrease the incidence of pneumonia, intensive care unit stay or postoperative hospital stay, and did not increase the incidence of complications such as anastomotic leakage, recurrent nerve palsy, wound infection, sepsis and catheter-related fever. CONCLUSION: A neutrophil elastase inhibitor is beneficial in patients undergoing esophagectomy. More high quality, large sample, multi-center and randomized controlled trials are needed to validate this effect.     

中性粒细胞弹性蛋白酶抑制剂对慢性炎症气道高分泌的干预作用

目的 了解慢性炎症气道黏液高分泌发生的分子机制。方法 利用大鼠慢支炎模型，对其进行中性粒细胞弹性蛋白酶(NE)抑制剂-Sivelestat干预后，采用酶联免疫吸附试验(ELISA)及原位杂交方法分别检测支气管肺泡灌洗液(BALF)中黏蛋白(MUC)含量以及肺组织粘蛋白基因MUC5AC转录水平的表达。结果 Sivelestat明显降低慢性气道炎症大鼠气道黏蛋白含量，并从转录水平降低MUC的分泌。结论 NE是炎症气道黏液高分泌发生的重要始动因素。NE抑制剂在慢性阻塞性肺病的治疗中表现出潜在的临床应用价值。     

华法林钠治疗大鼠实验性结肠炎的作用

目的:探讨炎症性肠病中凝血异常与炎性反应的关系.明确华法林钠抗凝治疗在2,4,6-三硝基苯磺酸钠(TNBS)诱导的实验性大鼠结肠炎模型中的作用.方法:40只大鼠均分为正常对照组、结肠炎组、华法林钠组和柳氮磺胺吡啶(SASP)组,正常对照组予以0.9%氯化钠溶液灌肠,其余三组予以含有20 mg TNBS的50%乙醇溶液0.5 ml灌肠.造模后第7天,华法林钠组给予华法林钠溶液240 ng·kg-1·d-1灌胃;SASP组给予SASP溶液100 mg·kg-1·d<-1>灌胃;正常对照组和结肠炎组予以每天1次0.9%氯化钠溶液灌胃.第14天取大鼠血液和结肠后处死.比较各组大鼠疾病活动指数(DAI)、大鼠结肠大体评分、结肠组织病理评分、血清肿瘤坏死因子(TNF)浓度、凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)、血小板计数(PLT)和抗凝血酶活性(AT)值.结果:第14天华法林钠组和SASP组DAI值分别为1.20±0.45和1.78±0.90,均较结肠炎组(2.25±0.89)降低,但差异均无统计学意义(P值均>0.05).华法林钠组和SASP组大体评分分别为1.40±0.55和3.14±1.46,均低于结肠炎组(4.75±1.66,P值均<0.01);而华法林钠组更低于SASP组(P<0.01).病理评分华法林钠组和SASP组分别为4.00±1.41和4.28±1.49,与结肠炎组比较差异有统计学意义(7.75±1. 04,P值均<0:01).血清TNF-α正常对照组较其余三组低(P<0.01),而结肠炎组较其余三组高(P<0.01).结肠炎组PT、APTT较华法林钠组和正常对照组缩短,AT活性也较后两组低(P值均<0.01).结肠炎组血小板计数明显高于正常对照组、SASP组和华法林钠组(P值均<0.01).结论:TNBS诱导的大鼠结肠炎模型中存在凝血异常.华法林钠治疗可减轻实验性大鼠结肠炎的炎症和损伤水平.