复方银黄微型灌肠剂对发热大鼠下丘脑中COX-1、COX-2 mRNA表达的影响

目的 研究复方银黄微型灌肠剂(YHCE)对发热大鼠下丘脑中环氧化酶-1(COX-1)、环氧化酶-2(COX-2)mRNA表达的影响,探讨其作用机制.方法 用酵母混悬液复制发热大鼠模型,采用实时荧光定量聚合酶链反应检测大鼠下丘脑中COX-1、COX-2的mRNA表达的变化.结果 YHCE能明显抑制发热大鼠的体温上升,抑制下丘脑中COX-2的mRNA表达,而对下丘脑中COX-1的mRNA无明显影响.结论 YHCE有明显的解热作用,其机制可能与抑制发热大鼠下丘脑中COX-2的mRNA表达有关.     

环氧化酶-2抑制剂体外筛选模型在药物抗炎机制研究中的应用

目的考察化合物ZA-76和中药单体LM、XR和HQ对环氧化酶-2(COX-2)的影响。方法利用作者建立的COX-2抑制剂体外筛选模型。结果发现化合物ZA-76对COX-2有较强抑制作用，中药单体LM、XR对COX-2有一定的抑制作用，HQ对COX-2有较弱抑制作用。结论可以应用模型进行COX-2抑制剂的筛选，并能阐明部分抗炎药物的作用机制。     

豨莶草对黄嘌呤氧化酶和环氧合酶-2双重抑制作用的虚拟筛选

目的 探究豨莶草对黄嘌呤氧化酶（XOD）和环氧合酶-2（COX-2）具有双抑制活性成分及结合机制。方法 由TCMSP数据库筛选得出豨莶草所含化合物成分,以XOD、COX-2为目标,使用Autodock Tools软件对化合物进行虚拟筛选,并使用Pymol软件针对代表性活性成分与黄嘌呤氧化酶、环氧合酶作用模式进行分析。结果 豨莶草所含的活性成分与XOD的结合能,在-5.0 kcal·moL^-1以下的有5种化合物,与COX-2的结合能,在-5.0 kcal·moL^-1以下的有9种化合物。根据分子间作用力来看,豨莶草中的豆甾醇、豨莶精醇对这两种蛋白的活性位点有氢键相互作用和疏水相互作用。结论 豨莶草中的豆甾醇、豨莶精醇成分可能对XOD和COX-2有双重抑制的效果。     

环氧化酶-2抑制剂防治肝癌的研究进展

目的:介绍环氧化酶-2(COX-2)抑制剂预防和治疗肝癌的研究进展及其作用机制.方法:查阅近年来国内外相关文献.结果:COX-2抑制剂在体内和体外试验中均能显著抑制多种肝癌细胞的增殖,诱导其凋亡,并对肿瘤血管生成有抑制作用.结论:COX-2可能成为肿瘤治疗中的一个新靶点,COX-2抑制剂在肝癌的防治中有较好的应用前景,但其有效性和安全性还需进一步研究.     

COX-2在变应性鼻炎患者血清中的表达

目的 检测变应性鼻炎患者与正常人群血清环氧化酶-2的水平,探讨血清环氧化酶-2与变应性鼻炎的关系.方法 采用酶联免疫吸附试验(ELISA法)检测变应性鼻炎患者与正常人的血清COX-2水平,比较AR患者治疗前后与健康人群中血清COX-2浓度的差异.结果 AR组患者治疗前后血清COX-2水平均高于正常对照组血清COX-2水平,有统计学意义(P＜0.05);AR患者治疗后血清COX-2水平与对照组比较,无统计学意义(P＞0.05).结论 血清COX-2可能与变应性鼻炎发生有关,有望成为AR患者疾病发生、发展、治疗及预后的动态观察指标.     

补阳还五汤对家兔血管内皮损伤后细胞间黏附分子-1及环氧化酶-1表达的影响

目的 探讨中药补阳还五汤对家兔血管内皮损伤细胞间黏附分子-1( ICAM-1)和环氧化酶-1( COX-1)表达的影响.方法 将健康新西兰大耳白兔20只,随机分为模型组、中药组各10只,模型组、中药组高脂饲料喂养4周后行腹主动脉内膜剥脱术造模,术后各组予正常饲料喂养,中药组予补阳还五汤灌胃4周,模型组予0.9％氯化钠注射液灌胃4周,造模术4周后处死家兔取腹主动脉,用免疫组化法检测各组对ICAM-1和COX-1表达的影响.结果 免疫组化染色显示,中药组ICAM-1表达(3.062±0.056)(A值)较模型组的(6.285±0.242)(A值)差异有统计学意义(t=14.41,P＜0.01);模型组COX-1蛋白表达(0.125±0.021)(A值)显著高于中药组的(0.068±0.013)(A值)(t=12.14,P ＜0.01).结论 补阳还五汤对血管内膜增殖早期ICAM-1和COX-1有拮抗作用.     

环氧化酶-2在大肠癌中的表达及其与Bcl-2的相关性研究

目的 通过观察环氧化酶-2(COX-2)和Bcl-2在大肠癌中的表达及其与大肠癌生物学特性的关系,初步探讨二者在大肠癌发生、发展过程中的作用机制.方法 应用免疫组织化学染色法对78例大肠癌组织、21例大肠腺瘤组织和13例正常大肠粘膜组织进行免疫组化染色.结果 (1)COX-2与Bcl-2均在大肠腺瘤及大肠癌中高表达.(2)COX-2表达与肿瘤的Duke's分期、分化程度、淋巴结转移有相关性,而Bcl-2的表达与肿瘤的Duke's分期、淋巴结转移有相关性. (3)COX-2的表达与Bcl-2的表达呈正相关性.结论 COX-2在大肠癌组织中的表达与Bcl-2呈正相关,提示COX-2可能通过上调Bcl-2途径抑制细胞凋亡而产生致癌作用.     

基质金属蛋白酶-2和环氧化酶-2在大肠腺癌中的表达水平及临床意义

目的 探讨基质金属蛋白酶-2(MMP-2)和环氧化酶-2(COX-2)在大肠腺癌中的表达水平及临床意义.方法 选取2012年10月-2013年11月符合研究标准的大肠腺癌患者55例,收集患者的临床资料、病理学资料等进行回顾性分析.另选同期正常大肠黏膜病理标本11例,使用免疫组织化学SP法检测各标本中MMP-2和COX-2的阳性表达率,分析MMP-2和COX-2的表达水平与病理特征的关系及临床意义.结果 大肠腺癌COX-2和MMP-2阳性表达率高于正常大肠黏膜(P ＜0.001).COX-2阳性表达与大肠腺癌患者的性别、年龄、肿瘤大小、大体类型、分化程度、浸润深度、淋巴结转移和TNM分期无相关性(P＞0.05);MMP-2阳性表达与大肠腺癌患者的分化程度具有相关性(P＜0.05).结论 大肠腺癌组织的MMP-2和COX-2表达均上调,MMP-2分子表达水平可提示患者病情进展程度.     

二氢青蒿素联合环氧化酶2抑制剂抗S180肉瘤作用机制研究

目的探讨二氢青蒿素联合环氧化酶2抑制剂对S180肉瘤的生长抑制作用及其可能的机制。方法采用ICR小鼠皮下移植瘤模型,观察二氢青蒿素联合环氧化酶2抑制剂对小鼠S180肉瘤生长抑制作用,以及对肿瘤COX-2、VEGF、CD31蛋白表达与白细胞数目的影响。结果二氢青蒿素分别以50、100、150mg·kg-1剂量与环氧化酶2抑制剂美洛昔康10mg·kg-1剂量合用能明显抑制S180肉瘤的生长,抑瘤率最高达55.38%,优于两药单用的效果;免疫组织化学检测表明,3个剂量的二氢青蒿素合用环氧化酶2抑制剂的肿瘤组织COX-2、VEGF及CD31蛋白的表达量均下降;Westernblot方法检测得在高剂量二氢青蒿素(150mg.kg-1.d-1)联合应用美洛昔康(10mg.kg-1.d-1)组VEGF和COX-2蛋白的表达量均有下降,与模型对照组差异有显著性(P<0.05);RT-PCR检测得各合用药物组的肿瘤组织VEGFmRNA表达量均减少(P<0.05)。结论二氢青蒿素联合环氧化酶2抑制剂能明显抑制S180肉瘤的生长,其机制可能与下调肿瘤组织中COX-2和VEGF的表达有关。     

环氧化酶-2和Fas在宫颈鳞癌中的表达及意义

目的 探讨环氧化酶-2(COX-2)和Fas在宫颈鳞癌组织中的表达及临床意义.方法 采用免疫组织化学方法,检测COX-2和Fas在45例宫颈癌和25例正常宫颈组织中的表达水平.结果 宫颈鳞癌COX-2的表达率(64.4%,29/45)明显高于正常宫颈组织,差异有统计学意义(P<0.01);Fas的表达率(42.2%,19/45)明显低于正常宫颈组织(84.0%,21/25),差异有统计学意义(P<0.01).COX-2、Fas的表达与宫颈鳞癌的病理分级、国际妇产科联盟(FIGO)分期无关(P>0.05),有淋巴结转移COX-2的表达明显高于无淋巴结转移(P<0.05);有淋巴结转移Fas的表达明显低于无淋巴结转移癌(P<0.05).COX-2与Fas的表达呈负相关(rs=-0.401,P<0.05).结论 COX-2和Fas的过表达可能参与了宫颈鳞癌的发生和发展.     

Quantitative structure–activity relationship studies of cyclooxygenase inhibitors: a comprehensive analysis

A quantitative structure‐activity relationship (QSAR) analysis of 10 structurally diverse set of compounds recently reported as cyclooxygenase (COX) inhibitors has been performed using ClogP, CMR, aromatic substituent constants, and suitable indicator variables. These revealed several important physicochemical and structural requirements for COX‐1, COX‐2 inhibitory activity, and selective inhibition of COX‐2 versus COX‐1 among these novel ligands. Seventeen QSAR models reported herein provide interesting insights in understanding the hydrophobic, steric, electronic, and structural requirements of COX inhibition among these individual set of compounds. These results may be used to further the design and development of selective COX‐2 inhibitors among these newly reported COX inhibitors. Drug Dev Res 64:220–231, 2005. © 2005 Wiley‐Liss, Inc.     

Trimethyl‐4‐oxo‐4,5,6,7‐tetrahydroindazole‐1‐acetic Acid: A New Lead Compound with Selective COX‐2 Inhibitory Activity

A novel series of 3,6,6‐trimethyl‐4‐oxo‐4,5,6,7‐tetrahydroindazole‐1‐acetic acid derivatives was designed and synthesized by a new one‐step pathway. Structure elucidation of the synthesized compounds was confirmed by various spectral and elemental analyses. The prepared compounds were evaluated for their ability to inhibit cyclooxygenase‐2 (COX‐2) and cyclooxygenase‐1 (COX‐1) enzymes in vitro. Among the synthesized compounds, the 2‐(3,6,6‐trimethyl‐4‐oxo‐4,5,6,7‐tetrahydroindazol‐1‐yl)acetic acid 4 emerged as the most potent COX‐2 inhibitor (IC₅₀ value: 150 nM) with the highest selectivity index (COX‐1/COX‐2 inhibition ratio: 570.6). Docking studies of compound 4 in the active site of COX‐2 recognized its potential binding mode to the enzyme. Based on the preliminary results, compound 4 was considered as a lead compound for further optimization.     

环氧合酶结合抑制剂的分子图形学研究

环氧合酶具有两种结构亚型,即环氧合酶－１和环氧合酶－２,环氧合酶－２选择性抑制剂的毒副作用低,是较好的非甾体抗炎药物。本文通过选择性抑制剂ＳＣ－５５８和非选择性抑制剂吲哚美辛分别与环氧合酶－１和２－进行对接,利用分子图形学,从定量和定性的角度阐明了抑制剂选择结合环氧合酶－１和２－的原因。     

Amide derivatives of [6-(5-methyl-3-phenylpyrazole-1-yl)-3(2H)-pyridazinone-2-yl]acetic acids as potential analgesic and anti-inflammatory compounds

In this study, amide derivatives of [6-(5-methyl-3-phenyl-pyrazole-1-yl)-3(2H)-pyridazinone-2-yl]acetic acid were synthesized and tested for their in vivo analgesic and anti-inflammatory activity by using the p-benzoquinone-induced writhing test and carrageenan-induced hind paw edema model, respectively. The analgesic and anti-inflammatory activity of the compounds 6a, 6d, 6e, 6g, 6h and 6m were more potent than that of aspirin as an analgesic and indomethacin as an anti-inflammatory drug, respectively. The other derivatives generally resulted in comparable activity to reference compounds. Inhibitor activity of the active compounds on cyclooxygenase isoforms was also investigated by using in vitro human whole blood assay and found that these derivatives did not exert their analgesic and anti-inflammatory activities through COX inhibition and other mechanisms might be involved.     

Comparison of cyclooxygenase inhibitory activity and ocular anti-inflammatory effects of ketorolac tromethamine and bromfenac sodium

ABSTRACT

Objective: To compare the cyclooxygenase (COX) activity and anti-inflammatory effects of the nonsteroidal anti-inflammatory drugs (NSAIDs) ketorolac tromethamine (ketorolac) and bromfenac sodium (bromfenac).

Methods: Cyclooxygenase activity and selectivity was determined in vitro by measuring prostaglandin E₂ (PGE₂) production following incubation of varying concentrations of NSAID with human recombinant COX‐1 or COX‐2 and arachidonic acid. Anti-inflammatory effects were evaluated in a rabbit model in which an ocular inflammatory response was induced by intravenous injection of 10?µg/kg lipopolysaccharide (LPS). In study animals, one eye was treated with 50?µL (+/–) ketorolac 0.4% (Acular LS) or bromfenac 0.09% (Xibrom) and the other eye with 50?µL buffered saline. In control animals, both eyes were treated with vehicle. All animals were treated twice: 2 hours and 1 hour before LPS.

Main outcome measures: PGE₂ production in vitro, measured by enzyme immunoassay; fluorescein isothiocyanate (FITC)-dextran leakage into the anterior chamber, measured by fluorophotometry; aqueous PGE₂ levels in vivo, measured by ELISA immunoassay.

Results: Ketorolac was six times more active against COX‐1 (?IC₅₀ = 0.02?µM) than COX‐2 (?IC₅₀ = 0.12?µM) while bromfenac was ≈ 32 times more active against COX‐2 (?IC₅₀ = 0.0066?µM) than COX‐1 (?IC₅₀ = 0.210?µM). In the animal model, both drugs resulted in nearly complete inhibition of FITC-dextran leakage and PGE₂ production in the anterior chamber of treated eyes. There was also a 79% inhibition (?p < 0.001) of FITC-dextran leakage in the contralateral eyes of bromfenac-treated rabbits, and a 22.5% inhibition (not statistically significant) in the contralateral eyes of ketorolac-treated rabbits.

Conclusions: Ketorolac is relatively COX‐1 selective while bromfenac is potently selective for COX‐2 over COX‐1. In the animal model, both ketorolac 0.4% and bromfenac 0.09% demonstrated maximal anti-inflammatory activity in treated eyes. Only bromfenac 0.09% had a significant effect on the contralateral eye, suggesting possible systemic absorption of this drug.     

Update on cyclooxygenase inhibitors: has a third COX isoform entered the fray?

ABSTRACT

It has been more than 30 years since Sir John Vane first reported that the pharmacological actions of aspirin-like drugs could be explained by their ability to inhibit cyclooxygenase (COX). Since then, a second isoform of COX, named COX‐2, has been discovered and highly selective inhibitors of this isoform have been marketed. Most recently, a splice variant of COX‐1 mRNA, retaining intron 1, and given the names COX‐3, COX‐1b or COX‐1v, has been described.

Non‐selective NSAIDs such as ibuprofen and naproxen, which inhibit both COX‐1 and COX‐2, have proven highly effective and safe in the short-term management of acute pain. Highly selective COX‐2 inhibitors including celecoxib, rofecoxib, valdecoxib, lumiracoxib, and etoricoxib were developed with the hope of significantly reducing the serious gastrointestinal toxicities associated with chronic high‐dose NSAID use. While long-term studies demonstrated that rofecoxib and lumiracoxib reduced the incidence of GI perforations, ulcerations and bleeds by approximately 60% compared to non-selective NSAIDs, recent reports also demonstrated that the chronic use of rofecoxib and celecoxib in arthritis and colorectal polyp patients, and the short-term use of parecoxib and valdecoxib in patients who had undergone coronary artery bypass surgery, resulted in a significant increase in serious cardiovascular events, including myocardial infarction and stroke compared to naproxen or placebo.

COX‐3 mRNA has been isolated in many tissues including canine and human cerebral cortex, human aorta, and rodent cerebral endothelium, heart, kidney and neuronal tissues. In transfected insect cells, canine COX‐3 protein is expressed and was selectively inhibited by acetaminophen. However, in humans and rodents an acetaminophen sensitive COX‐3 protein is not expressed because the retention of intron‐1 adds 94 and 98 nucleotides to the COX‐3 mRNA structure respectively. Since the genetic code is a triplicate code (3 nucleotides to form one amino acid), the retention of the intron in both species results in a frame shift in the RNA message and the production of a truncated protein with a completely different amino acid sequence than COX‐1 or COX‐2 lacking acetaminophen sensitivity.

Advances made through a combination of basic molecular biological and pharmacological techniques, and well designed randomized controlled clinical trials have demonstrated that the apparent gastrointestinal advantage of selective COX‐2 inhibitors appears to be outweighed by their potential for cardiovascular toxicity and that acetaminophen's analgesic and antipyretic effects do not involve the inhibition of the COX-1 splice variant protein, putative COX‐3.     

非甾体抗炎药物及其作用机理进展

通过对大量文献的调研 ,总结了花生四稀酸代谢的主要途径 ,以及代谢产物与炎症发生和发展的关系。综述了新结构类型抗炎镇痛药或化合物的主要特点。介绍了环氧化酶公知的两种亚型COX 1与COX 2。指出了COX 2是非甾体抗炎药物治疗作用的靶酶。预见了COX 2抑制剂 ,CO/ 5 LO双重抑制剂 ,特异性 5 LO抑制剂应是未来抗炎药研究的重点方向     

Phthalate plasticizer di(2‐ethyl‐hexyl) phthalate induces cyclooxygenase‐2 expression in gastric adenocarcinoma cells

The phthalate plasticizer, di(2‐ethyl‐hexyl) phthalate (DEHP), and its derived metabolites are common anthropogenic environmental toxins, which are known to act as endocrine disruptors. Numerous studies have associated DEHP with disruption of sex hormones, abnormal development of reproductive organs, allergies, and inflammation. Its role in promoting inflammation has been reported by both human epidemiological and animal studies. In stomach tissue, chronic inflammation is known to accompany mucosal damage, and pave the way to gastritis, stomach ulcers, and ultimately gastric cancer. Eastern Asian populations possess the highest gastric cancer incidences in the world. Coincidentally, East Asia is one of the world's major sites for plastics manufacture and export. Thus, possible correlations between DEHP, a common plasticizer, and gastric cancer are of great interest. Our study revealed several critical findings. First, even at very low dosage, mimicking the residual plasticizer exposure, detrimental effects of DEHP on gastric cells can be detected. Second, gastric cells treated with DEHP increased cyclooxygenase‐2 (COX‐2) in a time‐dependent manner. Third, promoter deletion studies revealed a critical role of nuclear factor‐kappa B (NF‐κB) for COX‐2 gene responses. Finally, our results indicated that a low concentration of DEHP is able to trigger COX‐2 activation via the extracellular signal–regulated kinase (ERK1/2) and NF‐κB signaling pathway. Taken together, we demonstrate that very low doses of DEHP enhance the expression of the prototypical inflammatory gene, COX‐2, in gastric cancer cells via ERK1/2 and NF‐κB activation. This study provides important insights into the inflammatory process and damages associated with phthalate plasticizers exposure.     

Paracetamol – An old drug with new mechanisms of action

Paracetamol (acetaminophen) is the most commonly used over‐the‐counter (OTC) drug in the world. Despite its popularity and use for many years, the safety of its application and its mechanism of action are still unclear. Currently, it is believed that paracetamol is a multidirectional drug and at least several metabolic pathways are involved in its analgesic and antipyretic action. The mechanism of paracetamol action consists in inhibition of cyclooxygenases (COX‐1, COX‐2, and COX‐3) and involvement in the endocannabinoid system and serotonergic pathways. Additionally, paracetamol influences transient receptor potential (TRP) channels and voltage‐gated Kv7 potassium channels and inhibits T‐type Cav3.2 calcium channels. It also exerts an impact on L‐arginine in the nitric oxide (NO) synthesis pathway. However, not all of these effects have been clearly confirmed. Therefore, the aim of our paper was to summarize the current state of knowledge of the mechanism of paracetamol action with special attention to its safety concerns.     

Synthesis of Novel Substituted Thiourea and Benzimidazole Derivatives Containing a Pyrazolone Ring as Anti‐Inflammatory Agents

Two series of new 1‐(alkyl/aryl)‐3‐{2‐[(5‐oxo‐4,5‐dihydro‐1H‐pyrazol‐3‐yl)amino]phenyl}thioureas 2a – h and 5‐[2‐(substituted amino)‐1H‐benzimidazol‐1‐yl]‐4H‐pyrazol‐3‐ols 3a – i were designed and synthesized as anti‐inflammatory agents. The cyclooxygenase inhibitory activity of the newly synthesized compounds was investigated. All the compounds showed non‐selective inhibition of COX‐1 and COX‐2 enzymes which was consistent with their docking results. Compounds 2c , 2f , 2g , 3b , and 3g that showed the highest COX‐2 inhibitory activity were selected for further in vivo testing as anti‐inflammatory agents using diclofenac as a reference drug. Two of the test compounds ( 2c and 3b ) showed potent anti‐inflammatory activity comparable to that of diclofenac with lower ulcerogenic effect relative to indomethacin. SAR study of the two series as cyclooxygenase inhibitors and anti‐inflammatory agents was also provided.