在患有急性肺损伤的小猪的肺泡巨噬细胞中一氧化氮，表面活性剂和糖皮质激素对核因子-κB和激活蛋白-1的活性的调控作用

AIM： To investigate whether acute lung injury (ALI) in ventilated piglets with bacterial infection affects NF-κB and AP-1 expression in alveolar macrophages (AM) and whether nitric oxide (NO), surfactant (Surf), glucocorticoids (GC) affect NF-κB and AP-1 activation in AM in vivo and in vitro. METHODS: The animals were intraperitoneally injected Escherichia coli, which caused ALI. Nuclear extracts of AM were analyzed by electrophoretic mobility shift assay (EMSA) for the nuclear factor-kappa B (NF-κB) and activation protein-1 (AP-1) expression. Detection of IκB-α protein was from cytoplasmic extract by Western blotting. Immunocytochemistry staining was used for intracellular location of p65 subunits of NF-κB. RESULTS: In ex vivo experiments, strikingly higher expression of NF-κB and AP-1 by EMSA was found 6h after bacterial injection in contrast to the Normal group. In the NO, SNO,and GC groups, markedly attenuated NF-κB and AP-1 activation was observed. The NF-κB and AP-1 activation in Surf group showed lower levels of the expression. Immunoblotting of AM cytoplasmic extract showed low expression of IκB-α protein in the Control and Surf groups. The stronger expression was observed in the NO, GC,and SNO groups. AM of the Control and Surf groups showed intense nuclear staining, with decreased nuclear staining in the NO, GC and SNO groups. In in vitro experiment, it caused a significant increase in NF-κB and AP1 activity in AM 1h after exposure to lipopolysaccharides (LPS). In AM treated by LPS SNP and LPS GC, all showed decrease of DNA binding activity of NF-κB and AP-1 compared to those exposed to LPS Surf. Immunoblotting of AM cytoplasmic extract showed that LPS stimulation of AM resulted in the low expression of IκB-α protein, which was not observed in the presence of SNP and methylprednisolone. However, the surfact antdid not show such effect. LPS Surf-exposed AM had intense nuclear staining, whereas decreased nuclear staining in the LPS NO and LPS GC-treated cultures was found, confirming a decrease in NF-rd3 activity. CONCLUSION:Activation of NF-κB was found in AM of ventilated piglets with bacterial ALI. NO and GS could prevent NF-κB and AP-1 activation in vivo and in vitro. Surfactant has limited effects on NF-κB and AP-1 activity.     

肺泡巨噬细胞ADAR1基因在急性肺损伤中表达的变化

目的:观察肺泡巨噬细胞(alveolar macrophag-es,AMs)RNA腺苷脱氨酶1(adenosine deaminase act-ing on RNA,ADAR1)基因(ADAR1基因)在急性肺损伤中的变化,探讨该基因在急性肺损伤病理变化中可能发挥的作用。方法:采用气管内滴注脂多糖(li-popolysaccharides,LPS)的方法制备急性肺损伤模型,经瑞氏染色计数急性肺损伤大鼠肺泡灌洗液中AMs、中性粒细胞(polymorphonuclear leukocytes,PMNs)并计算各自所占比例;以RT-PCR方法观察ADAR1基因在急性肺损伤中表达的变化。结果:气管内滴注脂多糖(2.5 mg.kg-1)成功制备的急性肺损伤模型中,肺泡灌洗液中细胞总数随时间变化而变化,由对照组的1.74×106增加至炎症反应6 h时17.13×106,9 h时17.00×106。AMs所占细胞总数的比例则由98%下降为21%,而其绝对数量随时间的推移不断的上升,尤其以6 h和9 h为著。ADAR1基因在急性肺损伤肺泡巨噬细胞中呈现时间依赖性的升高。结论:内毒素诱导的急性肺损伤模型中,AMs绝对数量显著上升,且其中ADAR1基因表达呈时间依赖性的升高,提示ADAR1基因可能在急性肺损伤发病过程及病理发展中发挥了重要的作用。     

Inhibition of inducible nitric oxide synthase attenuates acute endotoxin-induced lung injury in rats

1. In the present study, we investigated the effects of the inducible nitric oxide (iNOS) inhibitors S-methylisothiourea (SMT) and l-N(6)-(1-iminoethyl)-lysine (l-Nil) on endotoxin-induced acute lung injury (ALI), as well as the associated physiological, biomedical and pathological changes, in anaesthetized Sprague-Dawley rats and in rat isolated perfused lungs. 2. Endotoxaemia was induced by an intravenous (i.v.) infusion of lipopolysaccharide (LPS; Escherichia coli 10 mg/kg). Lipopolysaccharide produced systemic hypotension and tachycardia. It also increased the lung weight/bodyweight ratio, lung weight gain, exhaled nitric oxide (NO), the protein concentration in bronchoalveolar lavage and microvascular permeability. 3. Following infusion of LPS, plasma nitrate/nitrite, methyl guanidine, pro-inflammatory cytokines (tumour necrosis factor-alpha and interleukin-1beta) were markedly elevated. Pathological examination revealed severe pulmonary oedema and inflammatory cell infiltration. Pretreatment with SMT (3 mg/kg, i.v.) or l-Nil (3 mg/kg, i.v.) significantly attenuated the LPS-induced changes and ALI. 4. The results suggest that the inflammatory responses and ALI following infusion of LPS are due to the production of NO, free radicals and pro-inflammatory cytokines through the iNOS system. Inhibition of iNOS is effective in mitigating the endotoxaemic changes and lung pathology. Inhibitors of iNOS may be potential therapeutic agents for clinical application in patients with acute respiratory distress syndrome.     

吸入一氧化氮及静注异丙酚治疗兔油酸型急性肺损伤

目的观察吸入NO、静注异丙酚改善兔油酸型急性肺损伤(ALI)的氧合效果.方法用60mg·kg-1油酸引发兔ALI后(0h),随机分组.模型组(n=5)单纯行机械通气;NO组(n=5)持续吸入20ppmNO;异丙酚组(n=5)静注异丙酚2mg·kg-1·h-1.观察基础状态、0h、2h的平均动脉压(SAP)、HR、动脉血气及肺功能变化.治疗结束后摄胸片,测定肺组织湿/干(W/D)比值.结果治疗2h后与模型组相比,NO组PaO2>95mmHg,(P＜0.01)、PaO2/FiO2>300,明显提高(P＜0.05)、Qs/Qt、W/D显著下降(P＜0.05),异丙酚组无改变.结论急性肺损伤早期吸入NO可以安全有效地改善肺循环,提高氧合,减轻肺水肿,延缓肺损伤进程;异丙酚改善ALI氧合效果不显著.     

酵母多糖对小鼠腹腔巨噬细胞产生一氧化氮和白细胞介素-1的影响

目的探讨酵母多糖对小鼠腹腔巨噬细胞产生一氧化氮 (NO)和白细胞介素 1(IL 1)的影响。方法将不同剂量的酵母多糖加入体外培养的小鼠腹腔巨噬细胞中 ,取细胞培养上清液根据Griess反应检测NO-2 的量 ,间接反映巨噬细胞产生NO的生成量 ,并用溴化四唑蓝 (MTT)比色法检测上清液中IL 1的生成量。结果酵母多糖可明显促进小鼠腹腔巨细胞产生NO和IL 1,NO的生成量呈现剂量依赖关系。结论酵母多糖可诱导小鼠腹腔巨噬细胞产生NO和IL 1,可能是酵母多糖调节机体免疫功能、杀伤病原微生物和抗肿瘤的重要途径     

氨基胍对脂多糖诱导的大鼠肺表面活性物质和肺泡巨噬细胞的影响

目的观察氨基胍(aminogunidine,AG)对脂多糖(1ipopolysaccharide,LPS)诱导的大鼠肺表面活性物质(pulmonary surfactant,PS)时程性变化的影响和对肺泡巨噬细胞(alveolar macrophage,AM)的影响,探讨AG对肺损伤组织的保护作用及其机制。方法试验一:健康♂SD大鼠随机分为对照组、模型组和AG治疗组。模型组、AG治疗组静脉注射LPS(5mg·kg-1)复制内毒素性肺损伤模型。LPS3h和6h后腹腔注射AG(100mg·kg-1,AG治疗组)和生理盐水(对照组及LPS组)。原位杂交法(ISH)和Western blot法分别测定肺组织肺表面活性物质蛋白A(SP-A)mRNA及蛋白水平;留取肺泡灌洗液(BALF),测定BALF中的总磷脂(TPL)和总蛋白(TP)。试验二:体外分离培养大鼠肺泡巨噬细胞,以LPS(终浓度10mg·L-1)损伤巨噬细胞,观察AG对肺泡巨噬细胞的影响。结果与对照组比较,大鼠肺损伤后SP-A mRNA及蛋白表达在LPS6h、9h后明显下降;BALF中TPL明显减少,TP增多。肺损伤3h用AG治疗3h后,SP-A mRNA阳性细胞表达及蛋白明显增强,BALF中TPL较LPS组相同时间点明显上升,TP降低,病理显示肺损伤减轻。体外实验显示,与正常对照组相比,LPS处理巨噬细胞后,细胞培养上清中NO、TNF-α、IL-6和LDH浓度明显增高,AG明显减少LPS所致的LDH和NO的释放,降低TNF-α和IL-6浓度。结论肺损伤后早期给予AG可通过增强肺表面活性物质表达减轻内毒素性肺损伤。AG可抑制LPS对巨噬细胞的作用。     

Nitric oxide modulates air embolism-induced lung injury in rats with normotension and hypertension

1. Air embolism the in lungs induces microvascular obstruction, mediator release and acute lung injury (ALI). Nitrite oxide (NO) plays protective and pathological roles in ALI produced by various causes, but its role in air embolism-induced ALI has not been fully investigated. 2. The purpose of the present investigation was to elucidate the involvement of NO and pro-inflammatory cytokines in the pathogenesis of ALI following air infusion into isolated perfused lungs from spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto (WKY) rats. 3. The extent of ALI was evaluated by changes in lung weight, Evans blue dye leakage, the protein concentration in the bronchoalveolar lavage and pathological examination. We also measured nitrite/nitrate (NO(x)), tumour necrosis factor (TNF)-alpha and interleukin (IL)-1beta concentrations in lung perfusate and determined cGMP in lung tissue. 4. The NO synthase (NOS) inhibitors N(G)-nitro-l-arginine methyl ester (l-NAME) and l-N(6)-(1-iminoethyl)-lysine (l-Nil), as well as the NO donors sodium nitroprusside (SNP) and s-nitroso-N-acetylpenicillamine (SNAP), were administered 30 min before air embolism at a concentration of 10(-3) mol/L in the lung perfusate. 5. Air embolism-induced ALI was enhanced by pretreatment with l-NAME or l-Nil, but was alleviated by SNP or SNAP pretreatment, in both SHR and WKY rats. In both SHR and WKY rats, AE elevated levels of NO(x) (2.6 and 28.7%, respectively), TNF-alpha (52.7 and 158.6%, respectively) and IL-1beta (108.4 and 224.1%, respectively) in the lung perfusate and cGMP levels in lung tissues (35.8 and 111.2%, respectively). Pretreatment with l-LAME or l-Nil exacerbated, whereas SNP or SNAP abrogated, the increases in these factors, except in the case of NO(x) (levels were decreased by l-LAME or l-Nil pretreatment and increased by SNP or SNAP pretreatment). 6. Air embolism caused increases in the lung weight (LW)/bodyweight ratio, LW gain, protein concentration in bronchoalveolar lavage and Evans blue dye leakage. These AE-induced changes were less in lungs isolated from SHR compared with normotensive WKY rats. 7. The results suggest that ALI and associated changes following air embolism in lungs isolated from SHR are less than those in WKY rats. Nitric oxide production through inducible NOS isoforms reduces air embolism-induced lung injury and associated changes. Spontaneously hypertensive rats appear to be more resistant than WKY rats to air embolism challenge.     

Cyclic helix B peptide alleviates sepsis-induced acute lung injury by downregulating NLRP3 inflammasome activation in alveolar macrophages

Acute lung injury (ALI) exhibits high clinical morbidity and mortality rates. Our previous study has indicated that the novel proteolysis-resistant cyclic helix B peptide (CHBP) exerts an anti-inflammatory effect in mice with AKI. In the present study, we evaluated the effect of CHBP in an in vivo sepsis-induced ALI model and in vitro using lipopolysaccharide (LPS) and ATP stimulated bone marrow-derived macrophages (BMDMs). For in vivo experiments, mice were randomly divided into three groups: 1) sham; 2) LPS; and 3) LPS + CHBP (n = 6). All relevant data were collected after 18 h. Following CHBP treatment, the lung function of the mice was significantly improved compared to the LPS group. CHBP administration inhibited interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α production at both the protein and mRNA levels. Additionally, following CHBP treatment, the population of pulmonary macrophages decreased. Simultaneously, the proportion of caspase-1-activated alveolar macrophages was also decreased after CHBP treatment. The protein levels of NLRP3 and cleaved caspase-1 were attenuated in the lung tissue following CHBP treatment. In in vitro experiments, CHBP treatment decreased NLRP3 inflammasome expression and downstream IL-1β secretion, consistent with the in vivo results. In addition, CHBP reversed nuclear factor (NF)-κB and I-κB phosphorylation with a significant dose-dependent effect. Therefore, these findings suggest the potential of CHBP as a therapeutic agent in sepsis-induced ALI owing to inhibition of the NLRP3 inflammasome via the NF-κB pathway in macrophages.     

大鼠 paralemmin-3基因重组慢病毒载体构建及其在肺泡巨噬细胞中的表达

目的 构建大鼠 paralemmin-3（PALM3）基因重组慢病毒载体,探讨转染大鼠肺泡巨噬细胞系 NR8383细胞的最佳感染复数（MOI）值及目的基因 PALM3的表达情况。方法 采用聚合酶链反应技术（PCR）扩增鼠 PALM3基因片段,并将其克隆到 pCV186-Cherry载体上;经 PCR及 DNA测序鉴定后,将重组质粒 pCV186-Cherry-PALM3、辅助包装质粒 pHelper 1.0与 pHelper 2.0共转染至 293T细胞进行重组慢病毒 lenti-CV186-Cherry-PALM3的包装,并用荧光标记法行滴度的测定。将不同 MOI值（0、1、10、20、50）的 lenti-CV186-Cherry-PALM3转染 NR8383细胞,依据转染效率得到最佳 MOI 值,以此 MOI 值感染 NR8383细胞,采用 Western blot 法检测目的基因的表达情况。结果 通过 PCR扩增获得了大小约 2 246 bp的目的基因片段,并成功连接到 pCV186-Cherry重组质粒上。PCR及 DNA测序结果证实重组质粒 pCV186-cherry-PALM3 携带正确的鼠 PALM3 基因并成功包装重组慢病毒颗粒。重组慢病毒 lentiCV186-Cherry-PALM3 的滴度测定为 3×108 TU/mL,感染 NR8383 的最佳 MOI 为 20,慢病毒 lenti-CV186-CherryPALM3 感染后 NR8383 细胞中 PALM3 的蛋白表达水平显著上调（P＜0.05）。结论 本实验成功构建了含大鼠PALM3基因的重组慢病毒载体 lenti-CV186-Cherry-PALM3,并能在 NR8383细胞中高效表达,为进一步研究 PALM3对肺泡巨噬细胞极化的影响奠定了基础。     

Distribution characteristics of clarithromycin and azithromycin, macrolide antimicrobial agents used for treatment of respiratory infections, in lung epithelial lining fluid and alveolar macrophages

The distribution characteristics of clarithromycin (CAM) and azithromycin (AZM), macrolide antimicrobial agents, in lung epithelial lining fluid (ELF) and alveolar macrophages (AMs) were evaluated. In the in vivo animal experiments, the time‐courses of the concentrations of CAM and AZM in ELF and AMs following oral administration (50 mg/kg) to rats were markedly higher than those in plasma, and the area under the drug concentration–time curve (AUC) ratios of ELF/plasma of CAM and AZM were 12 and 2.2, and the AUC ratios of AMs/ELF were 37 and 291, respectively. In the in vitro transport experiments, the basolateral‐to‐apical transport of CAM and AZM through model lung epithelial cell (Calu‐3) monolayers were greater than the apical‐to‐basolateral transport. MDR1 substrates reduced the basolateral‐to‐apical transport of CAM and AZM. In the in vitro uptake experiments, the intracellular concentrations of CAM and AZM in cultured AMs (NR8383) were greater than the extracellular concentrations. The uptake of CAM and AZM by NR8383 was inhibited by ATP depletors. These data suggest that the high distribution of CAM and AZM to AMs is due to the sustained distribution to ELF via MDR1 as well as the high uptake by the AMs themselves via active transport mechanisms. Copyright © 2011 John Wiley & Sons, Ltd.     

Effects of an endogenous nitric oxide synthase inhibitor on phorbol myristate acetate-induced acute lung injury in rats

1. In the present study, we determined whether the endogenous nitric oxide (NO) synthase (NOS) inhibitor Nomega-nitro-l-arginine methyl ester (l-NAME) could ameliorate the acute lung injury (ALI) induced by phorbol myristate acetate (PMA) in rat isolated lung. 2. Typical ALI was induced successfully by PMA during 60 min of observation. At 2 micro g/kg, PMA elicited a significant increase in microvascular permeability (measured using the capillary filtration coefficient Kfc), lung weight gain, lung weight/bodyweight ratio, pulmonary arterial pressure (PAP) and protein concentration of bronchoalveolar lavage fluid. 3. Pretreatment with the NOS inhibitor l-NAME (5 mmol/L) significantly attenuated ALI. None of the parameters reflective of lung injury showed significant increase, except for PAP (P < 0.001). The addition of l-arginine (4 mmol/L) blocked the protective effective of l-NAME. Pretreatment with l-arginine exacerbated PMA-induced lung injury. 4. These data suggest that l-NAME significantly ameliorates ALI induced by PMA in rats, indicating that endogenous NO plays a key role in the development of lung oedema in PMA-induced lung injury.     

Emodin ameliorates acute lung injury induced by severe acute pancreatitis through the up‐regulated expressions of AQP1 and AQP5 in lung

The present study investigates the ameliorating effects of emodin on acute lung injury (ALI) induced by severe acute pancreatitis (SAP). An ALI rat model was constructed by sodium ursodeoxycholate and they were divided into four groups: SHAM, ALI, emodin and dexamethasone (DEX) (n=24 per group). Blood samples and lung tissues were collected 6, 12 and 24 hours after the induction of SAP‐associated ALI. Lung wet/dry ratio, blood gases, serum amylase and tumor necrosis factor‐α (TNF‐α) were measured at each time point. The expressions of AQP1 and AQP5 in lung tissue were detected by immunohistochemical staining, western blotting and real‐time PCR. As the results show, there were no statistical differences in the levels of serum amylase, lung wet/dry ratio, blood gases indexes, serum TNF‐α and pathological changes between emodin and DEX groups. However, significant differences were observed when compared with the ALI group. AQP1 and AQP5 expressions were significantly increased and lung oedemas were alleviated with the treatment of emodin and DEX. The expressions of AQP1 and AQP5 were significantly decreased in SAP‐associated ALI rats. Emodin up‐regulated the expression of AQP1 and AQP5, it could reduce pulmonary oedema and ameliorate SAP‐induced ALI. Regulations on AQP1 and AQP5 expression had a great value in clinical application.     

Regulation of endothelial nitric oxide synthase and asymmetric dimethylarginine by matrine attenuates isoproterenol-induced acute myocardial injury in rats

Objectives This study was designed to investigate the cardioprotective effects of matrine on regulation of endothelial nitric oxide synthase (eNOS) and asymmetric dimethylarginine (ADMA) in isoproterenol‐induced acute myocardial ischaemic rats. Methods Male Sprague–Dawley rats were pretreated with matrine (200, 100 and 50 mg/kg) orally for 10 days. Acute myocardial injury was induced in rats by subcutaneous injection of isoproterenol. Serum and haemodynamic parameters, histopathological variables and expression of protein levels were analysed. Key findings Oral administration of matrine (200, 100 and 50 mg/kg) significantly attenuated isoproterenol‐induced cardiac necrosis and left ventricular dysfunction. Matrine treatment restored impaired ventricular Akt and eNOS protein expression with concomitant increased phosphorylation of Akt (Ser473) and eNOS (Ser1177), and also restored glycogen synthase kinase 3β activity, as indicated by increased phosphorylation at Ser 9. Moreover, treatment with matrine had no effect on the isoproterenol‐induced elevated protein arginine methyltransferase 1 protein expression, but could significantly normalize the reduced dimethylarginine dimethylaminohydrolase 2 expression and attenuate the increased serum level of ADMA. The expression of catechol‐o‐methyltransferase and monoamine oxidase did not differ among all groups (all P > 0.05). Conclusions Our results suggested that matrine protects against isoproterenol‐induced myocardial ischaemia via eNOS and ADMA pathway.     

姜黄素抑制HMGB1-NF-κB信号通路减轻脂多糖诱导新生大鼠急性肺损伤实验研究

目的 分析姜黄素抑制高迁移率族蛋白B1（HMGB1）-核转录因子κB（NF-κB）信号通路减轻脂多糖（LPS）诱导新生大鼠急性肺损伤（ALI）的作用。方法 将60只新生SD雄性大鼠随机分为对照组、模型组、地塞米松（阳性药,2 mg·kg^-1）组和姜黄素低、中、高剂量（1.5、3.0、6.0 mg·kg^-1）组,每组10只。除对照组外,所有大鼠采用腹膜内注射LPS （3 mg·kg^-1）建立ALI模型。注射LPS约6 h后开始ip给药,每天1次,连续7 d,模型组和对照组大鼠ip等体积的0.1% DMSO。通过血氧分压（PaO₂）和肺干湿质量比（W/D）评估新生大鼠肺水肿情况;HE染色检测各组大鼠肺组织损伤;ELISA法检测新生大鼠支气管肺泡灌洗液（BALF）氧化应激指标超氧化物歧化酶（SOD）、髓过氧化物酶（MPO）、丙二醛（MDA）和谷胱甘肽（GSH）水平,BALF中白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和HMGB1水平;Western blotting法检测大鼠肺组织胞核NF-κB、胞浆NF-κB和磷酸化核因子κB抑制因子α（p-IκBα）蛋白表达。结果 与对照组相比,模型组新生大鼠肺泡腔有渗出、肺组织结构紊乱、细胞核固缩深染、伴随大量的炎性细胞浸润,病理评分显著升高（P<0.01）; PaO₂、BALF中SOD和GSH水平显著降低（P<0.01）;肺W/D,BALF中MPO和MDA水平,BALF中IL-6、TNF-α和HMGB1水平,NF-κB胞核/胞浆比例和胞浆p-IκBα蛋白表达水平显著升高（P<0.01）。与模型组相比,姜黄素高、中剂量组和地塞米松组大鼠肺组织病理损伤减轻,肺泡腔渗出、炎性细胞浸润明显减少,病理评分显著降低（P<0.05、0.01）; PaO₂、BALF中SOD和GSH水平显著升高（P<0.05、0.01）;肺W/D,BALF中MPO、MDA水平,BALF中IL-6、TNF-α和HMGB1水平,NF-κB胞核/胞浆比例和胞浆p-IκBα蛋白表达水平显著降低（P<0.05、0.01）。结论 姜黄素可以通过抑制HMGB1-NF-κB信号通路减轻LPS诱导的新生大鼠ALI。     

YL-I-108, a synthetic chalcone derivative,inhibits lipopolysaccharide-stimulated nitric oxide production in RAW 264.7 murine macrophages: Involvement of heme oxygenase-1 induction and blockade of activator protein-1

Chalcones, a group of phenolic compounds, exhibit potent anti-inflammatory properties. In the present study, we synthesized chalcone derivative, YL-I-108 ((E)-1-(2-methoxy-4,6-bis(methoxymethoxy)phenyl)-3-(3-nitrophenyl)prop-2-en-1-one), and examined its effect on the production of pro-inflammatory mediators. Treatment of RAW 264.7 macrophages with YL-I-108 potently inhibited nitrite production stimulated by LPS. YL-I-108 treatment also markedly inhibited expressions of inducible nitric oxide synthase (iNOS) and tumor necrosis factor-α (TNF-α). Treatment of cells with YL-I-108 significantly inhibited LPS-stimulated activator protein-1 (AP-1)-dependent reporter gene expression, whereas nuclear factor-κB (NF-κB) activity was not affected, indicating that down-regulation of iNOS expression by YL-I-108 is attributed by blockade of AP-1. In addition, YL-I-108 treatment led to an increase in heme oxygenase-1 (HO-1) mRNA and protein expression, accompanied with the increased expression of nuclear factor-erythroid 2-related factor 2 (Nrf2). Treatment with SnPP, a selective HO-1 inhibitor, reversed YL-I-108-mediated suppression of nitrite production, suggesting that HO-1 induction is implicated in the suppression of NO production by YL-I-108. In contrast, SnPP treatment did not reverse YL-I-108-mediated suppression of AP-1 activation, suggesting that AP-1 inhibition by YL-I-108 is independent of HO-1 induction. Together, these results indicate that YL-I-108 suppresses NO production in LPS-stimulated macrophages via simultaneous induction of HO-1 expression and blockade of AP-1 activation.     

右美托咪啶预处理对下肢缺血再灌注性肺损伤及一氧化氮/内皮素-1失衡的影响

摘要： 目的 观察右美托咪啶预处理对止血带引起的肢体缺血/再灌注 （LIR） 致肺损伤的保护作用和对一氧化氮（NO） /内皮素 （ET） -1 失衡的影响。方法 选取 60 例 ASA 评分Ⅰ～Ⅱ级拟行单侧下肢手术的患者。随机分为对照组 （R 组, n=30） 和右美托咪啶预处理组 （PD 组, n=30）。所有患者均在神经刺激仪引导下行腰丛-坐骨神经阻滞麻醉后手术。PD 组于使用止血带前 10 min 静脉泵注右美托咪啶 0.125 mL/kg （4 mg/L）; R 组在相应时间点给予等量生理盐水 0.125 mL/kg。分别于使用止血带前 10 min（T0）和松止血带后 15 min（T1）、 2 h（T2）、 6 h（T3）及 24 h（T4）行动脉血气分析, 计算呼吸指数（RI）和氧和指数（OI）, 测定 NO、 ET-1、 白细胞介素（IL） -8、 丙二醛（MDA）水平。结果 与 T0 比较, R 组 T3时 RI 升高, T2~T4时 OI 降低 （P＜0.01）, PD 组各时点 RI、 OI 值比较差异无统计学意义; R 组和 PD 组在松止血带后 ET-1、 IL-8、 MDA 浓度升高, NO 浓度、 NO/ET-1 比值显著降低 （P＜0.05）。与 R 组比较, PD 组在 T3时 RI 降低、 OI 升高 （P＜0.01）; PD 组在松止血带后 ET-1、 IL-8、 MDA、 NO 浓度和 NO/ET-1 比值与 R 组比较差异有统计学意义（P＜0.05）。R 组患者血浆 ET-1、 IL-8 水平与 RI 值呈正相关, 与 OI 值呈负相关（P < 0.01）; 血浆 NO 水平、 NO/ ET-1 比值与 RI 值呈负相关, 与 OI 值呈正相关 （P < 0.01）。结论 右美托咪啶预处理通过保护内皮细胞和减轻脂质过氧化损伤, 改善患者使用止血带造成的 LIR 致肺换气功能的障碍。     

Altered expression of cyclooxygenase-2, 12-lipoxygenase,inducible nitric oxide synthase-2 and surfactant protein D in lungs of patients with pulmonary injury caused by sulfur mustard

Context: Sulfur mustard (SM) is a strong alkylating toxicant that targets different organs, particularly human lung tissue. Change in genes expression is one of the molecular mechanisms of SM toxicity in damaged tissue.

Objective: The purpose of this investigation is to characterize the expression of cyclooxygenase-2 (COX-2), 12-lipoxygenase (12-LO), inducible nitric oxide synthase 2 (iNOS2), and surfactant protein D (SFTPD) in lungs of patients who exposed to SM.

Methods: Lung biopsies were provided from SM-exposed patients (n?=?6) and controls (n?=?5). Total RNA were extracted from all specimens and then cDNA was synthesized for each sample. Changes in gene expression were measured using RT² Profiler ?PCR Array.

Results: Pulmonary function tests revealed more obstructive and restrictive spirometric patterns among patients compared to the control group. Expression of COX-2 and 12-LO in the lung of patients was increased by 6.2555 (p?=?0.004) and 6.2379-folds (p?=?0.002), respectively. In contrast, expression of SF-D and iNOS genes was reduced by 8.5869-fold (p?=?0.005) and 2.4466-folds (p?=?0.011), respectively.

Conclusions: Mustard lungs were associated with overexpression of COX-2 and 12-LO, which are responsible for inflammation, overproduction of free radicals and oxidative stress. Downregulation of iNOS2 and SF-D are probably the reason for lung disease and dysfunction among these patients. Therefore, the expression of these genes could be an important, routine part of the management of such patients.