Treatment of Chronic Obstructive Pulmonary Disease with Roflumilast,a New Phosphodiesterase 4 Inhibitor

ABSTRACT

Recent advances in chronic obstructive pulmonary disease (COPD) treatment offer symptom relief, but disease modification remains an unmet goal of pharmacotherapy. Reducing the frequency and severity of COPD exacerbations may help slow disease progression and reduce the morbidity, mortality, and costs associated with these major events. Other desirable characteristics for a COPD treatment include a once-daily dosing schedule, an oral formulation, and a low frequency of systemic side effects. Phosphodiesterase 4 inhibitors have been in clinical development for some years and roflumilast is currently the most advanced of these agents. In this review, the preclinical evidence, clinical safety, and efficacy of roflumilast available in published reports are considered. The data reviewed here suggest that the clinical efficacy of roflumilast occurs through a mechanism unrelated to bronchodilation and may be due to the suppression of lung inflammation. Lung function improved with roflumilast treatment and in some studies, the reduction in exacerbations was substantial and statistically significant. Notably, this effect appeared to be greatest in the subgroup of patients with more severe disease and more severe exacerbations. The evaluation of roflumilast safety largely centers on gastrointestinal adverse events, with diarrhea, nausea, and weight loss occurring more frequently with the drug than placebo. If approved for general use, we expect roflumilast to find its role initially as a substitute for inhaled corticosteroids in the maintenance treatment of severe and very severe disease, particularly in patients who have frequent acute exacerbations, and perhaps as a supplementary drug when symptoms are not adequately controlled by current conventional COPD therapy.     

Alterations of Adhesion Molecule Expression and Inflammatory Mediators in Acute Lung Injury Induced by Septic and Non-septic Challenges

The lung is frequently the first failing organ during the sequential development of multiple organ dysfunction under both septic or non-septic conditions. The present study compared polymorphisms of tumor necrosis factor (TNFα), monocyte chemoattractant protein-1 (MCP-1), and adhesion molecule (AM) expression on circulating, recruited, and migrating leukocytes in the development of lung injury after induction of acute pancreatitis (AP) or abdominal sepsis by cecal ligation and puncture (CLP). Pulmonary alveolar barrier and endothelial barrier permeability dysfunction were measured. The expression of AMs (CD11b, CD11b/c, CD31, CD54 and CD62L) on leukocytes isolated from blood, lung tissue, and bronchoalveolar space were measured by flowcytometry. Plasma exudation to the interstitial tissue and the bronchoalveolar space significantly increased 1 and 3 hours after induction of pancreatitis and to the bronchoalveolar space from 6 hours after sepsis. Bronchoalveolar levels of MCP-1 significantly increased earlier than plasma exudation to the alveoli in both pancreatitis and sepsis. Alterations in expression of adhesion molecules on bronchoalveolar lavage (BAL) leukocytes can represent a marker reflecting leukocyte activation in the lung tissue, since both BAL and lung tissue leukocytes showed similar patterns of changes. Expression of adhesion molecules on circulating leukocytes increased 1 hour after induction of pancreatitis. Activating phenotypes of circulating, lung tissue and bronchoalveolar leukocytes may thus be responsible for the-development and severity of secondary lung injury.Supported by grants from the Swedish Research Council (grant no 11236), the Crafoord Foundation, Ake Wiberg Foundation, Maj Bergvall Foundation, Golje Foundation and Clas Groschinsky Foundation.     

Nitro-Arginine Methyl Ester,a Non-Selective Inhibitor of Nitric Oxide Synthase Reduces Ibuprofen-Induced Gastric Mucosal Injury in the Rat

Ibuprofen is a commonly used non-steroidal anti-inflammatory drug. Gastrointestinal adverse drug reactions from ibuprofen usage include gastric mucosal ulcers and bleeding. The mechanism by which ibuprofen induces gastric mucosal damage is not clear. The present study is an attempt to examine the role of nitric oxide in the pathogenesis of ibuprofen-induced gastric mucosal damage. Ibuprofen administered orally at the dose of 100 mg/kg body weight for 6 days to the rats resulted in gastric mucosal injury. Serum nitrite and nitrosothiol were increased significantly as compared with the controls, which were treated with the vehicle alone. In the gastric mucosa, lipid peroxidation and protein thiols were increased, and the activity of glyceraldehyde 3-phosphate dehydrogenase, a nitric oxide sensitive enzyme was decreased significantly. Pretreatment of the rats daily with nitric oxide synthase inhibitor, nitro-arginine methyl ester (30 mg/kg body weight) 1 hr before treatment with ibuprofen reduced the gastric mucosal injury. Biochemically, it prevented the rise in serum nitrite levels and the increase in lipid peroxidation and protein thiol levels and the loss of glyceraldehyde 3-phosphate dehydrogenase activity in the gastric mucosa. The results of the present study suggest that increased nitric oxide production may be one of the mechanisms by which ibuprofen produces gastric mucosal injury and that inhibition of nitric oxide synthase reduces gastric mucosal injury.     

SOCS3在实验性急性胰腺炎急性肺损伤大鼠肺组织中的表达和作用

背景：细胞因子信号转导抑制分子3（SOCS3）在多种疾病的各种器官损伤中起重要的调节作用,可通过对JAK/STAT信号通路的负反馈作用调节炎症因子的释放,从而起一定的抑炎作用。目前关于SOCS3在重症急性胰腺炎（SAP）急性肺损伤中的表达和作用尚未见报道。目的：探讨SOCS3在实验性急性胰腺炎（AP）合并急性肺损伤大鼠肺组织中的表达变化及其可能的作用。方法：32只Sprague-Dawley大鼠随机分为对照组和AP 6 h、12 h、18 h组。以4%牛磺胆酸钠胰胆管逆行注射诱导AP模型。动态测定各组血清淀粉酶（AMY）水平、肺湿/干重比;光学显微镜下观察肺组织学表现;ELISA法检测血清白细胞介素（IL）-6、IL-18含量;免疫组化法和蛋白质印迹法检测肺组织中SOCS3的定位和表达。结果：与对照组相比,各AP模型组血清AMY水平、肺湿/干重比均明显升高（P〈0.05）;肺组织损伤随病情进展而逐渐加重;血清IL-6、IL-18水平显著上调（P〈0.05）;肺组织SOCS3表达逐渐增强（P〈0.05）,于18 h时达高峰。结论：SAP急性肺损伤导致的炎症反应可诱导SOCS3在肺组织中表达,并随着肺组织损伤和炎症反应严重程度的增加而逐渐增高,提示可能与其负反馈调节JAK/STAT信号通路介导的炎症反应的作用存在一定的联系。     

Dobutamine Enhances Alveolar Fluid Clearance in a Rat Model of Acute Lung Injury

Acute lung injury (ALI) and its most severe form, acute respiratory distress syndrome (ARDS), remain leading factors for morbidity and mortality in critically ill patients. A significant aspect of ALI and ARDS is impaired alveolar fluid clearance (AFC). Improvements in therapies for these types of respiratory illnesses will require an understanding of the mechanisms that control AFC. The present study was designed to determine whether the administration of dobutamine decreases pulmonary edema and stimulates AFC in a rat model of lipopolysaccharide-induced lung injury. Adult male Sprague–Dawley rats were randomly divided into three groups: control, lipopolysaccharide, and lipopolysaccharide + dobutamine. The effect of dobutamine on AFC and the expression of aquaporin-1 and aquaporin-5 were examined. Lipopolysaccharide administration results in significant lung injury with impaired AFC, while dobutamine improves alveolar fluid reabsorption with elevation of aquaporin-1 and aquaporin-5. Our study indicates that dobutamine may enhance alveolar fluid reabsorption by increasing the expression of aquaporin-1 and aquaporin-5.     

A Synthetic Selective Inhibitor of Factor Xa, DX-9065a, Reduces Monocyte Chemoattractant Protein-1 Expression After Ischemia-Reperfusion Injury in Rat Liver

Activated factor X (FXa) is a trypsinlike serineprotease involved in the cascade of blood coagulation.The monocyte chemoattractant protein-1 (MCP-1) may beimportant in the pathophysiology of liverischemia-reperfusion injury. We investigated the effects of aselective FXa inhibitor, DX-9065a, on MCP-1 expressionafter ischemia-reperfusion in the rat liver. Liverischemia was induced in rats by occluding the portalvein for 30 min. DX-9065a was injected intravenously5 min before vascular clamping. Serum concentrations ofMCP-1 were measured by enzyme-linked immunosorbentassay. The levels of MCP-1 mRNA in the liver after reperfusion were determined by northern blotanalysis. In vitro MCP-1 production by peritonealmacrophages in response to alpha-thrombin was examined.Serum concentrations of MCP-1 increased and peaked at 6 hr after reperfusion. However,pretreatment of animals with DX-9065a resulted insignificantly smaller increases in the serumconcentration of MCP-1 after reperfusion in adose-dependent manner. Pretreatment with DX-9065a significantly reduced MCP-1 mRNAlevels in the liver after ischemia-reperfusion. In vitroMCP-1 production by peritoneal macrophages was enhancedby alpha-thrombin. In addition, DX-9065a significantly reduced tissue factor mRNA levels in peripheralmonocytes after ischemiareperfusion, compared tountreated animals. In conclusion, a selective inhibitorof FXa, DX-9065a, limited MCP-1 production after ischemia-reperfusion of the ratliver.     

急性冠脉综合征患者pentraxin-3与凝血酶活化的纤溶抑制物的变化

目的了解急性冠脉综合征(Acute Coronary Syndrome,ACS)患者炎性指标pentraxin-3与纤溶指标凝血酶活化的纤溶抑制物(Thrombin activatable finolysis inhibitor,TAFI)的变化。方法检测比较102例急性心肌梗死(AMI组)、81例不稳定心绞痛(UAP组)及23例健康体检者(对照组)的血浆pentraxin-3、TAFI水平。结果急性冠脉综合征患者血浆pentraxin-3、TAFI明显高于对照组,差异有统计学意义(P<0.01);AMI组血浆pentraxin-3及TAFI水平高于UAP组,差异有统计学意义(P<0.05)。pentraxin-3浓度与TAFI浓度呈正相关(r=0.17,P<0.05)。结论 pentraxin-3与TAFI共同在ACS的发生发展中起重要作用。     

Antithrombotic Effects of BCH 2763, a New Direct Thrombin Inhibitor,in a Canine Model of Venous Thrombosis

Deep venous thrombosis (DVT) is a common cardiovascular disease, resulting in significant mortality each year in the United States. Direct thrombin inhibitors represent a new class of drugs that could potentially be better than conventional antithrombotic therapy based on indirect inhibition of coagulation factors with heparin and warfarin. BCH 2763 is a potent, selective bifunctional thrombin inhibitor that blocks both the active catalytic site and the anion binding exosite. The objective of this study is to test the antithrombotic efficacy of BCH 2763 in a canine model of DVT induced through electrolytic injury to the femoral vein. BCH 2763 was administered at three dose levels: 0.125 mg/kg bolus followed by 10 µg/kg/min IV infusion (low-dose; n = 5), 0.25 mg/kg bolus followed by 20 µg/kg/min infusion (mid-dose; n = 5), and 0.75 mg/kg bolus followed by 60 µg/kg/min (high-dose; n = 5). The control group (n = 5) received a 5-ml intravenous bolus of saline followed by a 1 mL/kg/h infusion. The parameters evaluated were changes in activated partial thromboplastin time (aPTT), thrombin time (TT), prothrombin time (PT), time to formation of an occlusive thrombus in the femoral vein, and the amount of venous blood flow delivered over the course of the experiment. There were significant dose-dependent increases in aPTT, TT, and PT in the BCH 2763-treated animals compared with the control group. The time to formation of an occlusive thrombus in the control group averaged 69.6 ± 9 minutes. Treatment with BCH 2763 prolonged the time to occlusion to 126.4 ± 13 minutes in the low-dose group, 155.4 ± 17 minutes in the mid-dose group, and 229 ± 7 minutes in the high-dose group (80% remained patent for the duration of the study), which were all significantly greater than the controls. Femoral venous blood flow was significantly greater in the mid-dose (51 ± 8%) and the high-dose (70 ± 6%) groups compared with the control vessels (22 ± 3%). In conclusion, the results of this study indicate that BCH 2763 is an effective intravenous antithrombotic agent in the canine electrolytic injury model of venous thrombosis.     

溃疡性结肠炎患者外周血单个核细胞中Foxp3 mRNA的表达水平

目的:探讨溃疡性结肠炎(UC)患者外周血单个核细胞中Foxp3 mRNA表达及其与疾病活动性的关系．方法:应用逆转录-聚合酶链反应(RT-PCR) 检测142例UC患者(活动期22例、缓解期20例) 和30例正常对照组外周血单个核细胞Foxp3 mRNA的表达水平．结果:UC患者急性期Foxp3 mRNA水平低于正常人,差异有显著性(1．58±0．31 vs 3．27± 0．40,P<0．05);缓解期Foxp3 mRNA水平与正常人差异无显著性(P=0．104);PBMC中Foxp3 mRNA表达水平与Walmsley评分标准有相关性,且呈负相关．结论:UC患者外周血单个核细胞中Foxp3 mRNA表达水平显著低于正常人,其参与了 UC的发病过程并与疾病的活动性有关．     

加味温胆汤对抑郁模型大鼠海马NMDA-NR1mRNA/蛋白表达的影响

目的探讨加味温胆汤对抑郁模型大鼠海马NMDA-NR1mRNA/蛋白表达的影响作用。方法以孤养结合慢性不可预见性应激抑郁模型大鼠为研究对象,在实验7d、14d、21d、28d时,检测各组大鼠旷场实验中水平运动、垂直运动得分,并采用QRT-PCR、Western-Blot技术检测各组大鼠海马NMDA-NR1mRNA/蛋白表达变化。结果 21d、28d时,与空白组比较,模型组水平运动、垂直运动得分降低(P<0.01);与模型组比较,中、西药组水平运动、垂直运动得分增加(P<0.05或P<0.01)。21d、28d时,与空白组比较,模型组大鼠海马NMDA-NR1mRNA/蛋白表达增多(P<0.01);与模型组比较,中、西药组大鼠海马NMDA-NR1mRNA/蛋白表达减少(P<0.05或P<0.01)。结论加味温胆汤的抗抑郁效应可通过阻抑海马NMDA-NR1mRNA/蛋白表达而实现。     

大鼠局灶性脑缺血再灌注后TNF-a、SOCS-3动态表达的实验研究

目的探讨肿瘤坏死因子-a（TNF—a）、细胞因子信号转导抑制因子-3（SOCS-3）在大鼠脑缺血再灌注后的动态表达及特点,阐明TNF-a、SOCS-3在脑缺血再灌注中的作用。方法将雄性sD大鼠48只随机分为假手术组和缺血3h再灌注3h、6h、12h、24h、48h、72h、7d组,6只／组。运用改良线栓法制作大鼠局灶性脑缺血再灌注模型。用放射免疫法测定TNF-a含量及用RT—PCR法测定SOCS-3的含量。结果TNF-a含量变化：与假手术组比较,缺血再灌注纽脑组织TNF—a含量在6h明显增加,12h达到高峰,随后各时间点表达开始下降。在缺血半暗带区,TNF—a含量变化与中心区相同,但含量较低。SOCS-3mRNA含量变化：缺血再灌注组缺血中心区SOCS-3表达在6h开始有明显升高与假手术组相比有显著性差异,再灌注24h时达到高峰,随后备时间点表达相对稳定,至再灌注7d时仍过分表达。在缺血半暗带区,SOCS～3mRNA变化与中心区相同,但含量较低。结论TNF—α在大鼠局灶性脑缺血再灌注损伤中具有重要作用,可诱导SOCS-3基因的产生和表达。而SOCS-3基因是一个脑缺血后上调基因,它参与了脑缺血再灌注损伤的全过程,可能起到了内源性神经保护剂的作用。     

系统性红斑狼疮外周血单个核细胞DcR3 mRNA表达研究

目的研究凋亡相关基因诱骗受体3(DcR3)在系统性红斑狼疮患者(SLE)外周血单个核细胞(PBMCs)中的mRNA表达水平。方法应用半定量逆转录-聚合酶链反应(RT-PCR)检测38例SLE患者(21例活动期,17例缓解期)和15例正常人PBMCs中DcR3 mRNA的表达水平,分析其与狼疮活动指数(SLEDAI)的相关性。结果SLE患者PBMCs中,DcR3mRNA表达水平明显高于正常人(P<0.01),但疾病不同病期(活动期与缓解期)其表达水平无统计学差异(P>0.05);DcR3 mRNA表达与SLEDAI不相关(r=-0.024,P>0.05)。结论SLE患者PBMCs DcR3 mRNA表达增加,提示DcR3可能与SLE免疫细胞凋亡异常和免疫调节的紊乱有关,参与了SLE的发病过程。     

Reduction of Plasma Cholesterol Levels and Induction of Hepatic LDL Receptor by Cerivastatin Sodium (CAS 143201-11-0, BAY w 6228), a New Inhibitor of 3-Hydroxy-3-methylglutaryl Coenzyme A Reductase, in Dogs

The effects of cerivastatin sodium (BAY w 6228), a new type of inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, on plasma cholesterol concentrations and the induction of hepatic LDL receptors were investigated with beagle dogs and Hep G2 cells. Oral administration of cerivastatin (0.01, 0.03, and 0.1 mg/kg per day) for 3 weeks reduced plasma total and very low-density lipoprotein plus low-density lipoprotein (VLDL + LDL) cholesterol concentrations and increased hepatic LDL receptor binding activity in dogs. Scatchard plot analysis revealed a 1.9-fold increase in the maximum binding capacity of hepatic LDL receptors in cerivastatin-treated animals. Similar results were obtained by administration of pravastatin (1.0 and 5.0 mg/kg/day) for 3 weeks. Binding activity of the LDL receptor, as well as receptor mRNA and protein concentrations, were increased in a dose-dependent manner (0.01–1.0 μM) by exposure of Hep G2 cells to cerivastatin. The results suggest that cerivastatin reduces plasma cholesterol concentrations by increasing hepatic LDL receptor expression. The mechanism of lowering cholesterol concentration by cerivastatin was the same as with the other previously examined HMG-CoA reductase inhibitors, but the effects with cerivastatin were apparent at doses much lower than the effective doses of the other drugs. Cerivastatin, therefore, shows potential for clinical use as a potent and efficacious plasma cholesterol-lowering drug. This revised version was published online in July 2006 with corrections to the Cover Date.     

压力性尿失禁患者阴道壁组织中MMP-10、TIMP-3和LN的表达及意义

目的观察女性压力性尿失禁（SUI）患者阴道壁组织中基质金属蛋白酶-10（MMP-10）及其抑制剂-3（TIMP-3）和层粘连蛋白（LN）的表达变化,并探讨其意义。方法随机选取40例有完整随访资料的女性SUI患者为SUI组,选取同期30例无压力性尿失禁和盆腔脏器脱垂（POP）的良性妇科病手术患者为对照组,均于术中取患者阴道壁结缔组织,采用免疫组化法检测其中的MMP-10、TIMP-3及LN。结果SUI组MMP-10的表达较对照组明显升高,TIMP-3和LN的表达较对照组明显下降（P〈0．05）。SUI组MMP-10与TIMP-3、LN的表达分别呈负相关（r分别为-0．494、-0．459,P均〈0．05）;TIMP-3与LN的表达呈正相关（r=0．946,P〈0．05）。结论SUI中MMP-10升高及TIMP-3和LN的下降,使胶原代谢活跃,对胶原纤维的降解明显增强。联合检测阴道壁组织中的MMP-10、TIMP-3和LN有助于SUI的诊疗。     

二烯丙基三硫化物包被支架术后冠状动脉管壁基质金属蛋白酶2及其抑制物表达的变化

目的探讨二烯丙基三硫化物包被支架对犬冠状动脉基质金属蛋白酶2及基质金属蛋白酶组织抑制物2表达的影响。方法利用犬冠状动脉置入过大支架致内膜损伤,建立血管狭窄的动物模型。28个支架平分为基础对照组(单纯蛋白包被支架)和二烯丙基三硫化物治疗组(二烯丙基三硫化物包被支架),分别置入14只杂种犬的冠状动脉前降支或回旋支,未置入支架的另一支冠状动脉作为正常对照组(n=14)。在不同时点(5 h,1、7、14天和28天)处死动物,术后4周的血管标本做病理切片,部分组织抽提总RNA,应用RT-PCR方法测定不同时点冠状动脉壁内基质金属蛋白酶2和基质金属蛋白酶组织抑制物2的表达。结果与基础对照组相比,4周后二烯丙基三硫化物包被支架组显著抑制新生内膜的形成(P<0.01),面积狭窄率显著降低(P<0.01);与基础对照组相比,在5 h、1、14天和28天二烯丙基三硫化物包被支架显著抑制冠状动脉壁内基质金属蛋白酶2 mRNA的表达(P<0.05)。在5 h、1天和14天3个时点,二烯丙基三硫化物组与基础对照组基质金属蛋白酶组织抑制物2的表达差异无显著性;在7天和28天,二烯丙基三硫化物组基质金属蛋白酶组织抑制物2的表达明显高于对照...     

Small-Angle X-ray Scattering Models of APOBEC3B Catalytic Domain in a Complex with a Single-Stranded DNA Inhibitor

In normal cells APOBEC3 (A3A-A3H) enzymes as part of the innate immune system deaminate cytosine to uracil on single-stranded DNA (ssDNA) to scramble DNA in order to give protection against a range of exogenous retroviruses, DNA-based parasites, and endogenous retroelements. However, some viruses and cancer cells use these enzymes, especially A3A and A3B, to escape the adaptive immune response and thereby lead to the evolution of drug resistance. We have synthesized first-in-class inhibitors featuring modified ssDNA. We present models based on small-angle X-ray scattering (SAXS) data that (1) confirm that the mode of binding of inhibitor to an active A3B C-terminal domain construct in the solution state is the same as the mode of binding substrate to inactive mutants of A3A and A3B revealed in X-ray crystal structures and (2) give insight into the disulfide-linked inactive dimer formed under the oxidizing conditions of purification.     

肺炎支原体感染小鼠Th1／Th2平衡状况及Tim-3mRNA的表达

目的通过检测Tim-3mRNA的表达水平及IFN-7、IL-4的含量,初步探讨肺炎支原体感染鼠Th1／Th2细胞的动态变化及Tim～3的免疫调控作用。方法通过滴鼻感染建立肺炎支原体肺炎小鼠模型,观察小鼠的一般状况,分别于感染后3d、5d、7d、14d、21d取材,应用RT—PCR方法检测肺组织和脾脏Tim-3mRNA的表达;采用ELISA法检测肺泡灌洗液中细胞因子IFN～γ、IL-4的含量。结果实验组IFN-γ于3d出现升高（t=2．309,P〈0．05）,5dIFNY开始下降,而IL-4于3d开始升高并一直持续到7d（t-6．436,P〈0．05）,之后逐渐恢复正常,肺组织和脾脏Tim-3mRNA的表达均于5d达到峰值（t=4．727,P〈0．05;t=7．962,P〈0．05）,且肺组织和脾脏Tim-3mRNA表达量与L-4呈正相关（r=0．561,P〈0．05;r=0．462,P〈0．05）,从5d开始与IFN-γ呈负相关（r=0．379,P〈0．05;r=0．431,P〈0．05）。结论小鼠感染肺炎支原体后Th1／Th2失衡,以Th2介导的免疫反应为主;Tim3可能参与了感染后对Th1细胞的负调控。