Wortmannin对急性肺损伤小鼠肺组织IL-1β和TNF-α表达的影响

目的研究Wortmannin对急性肺损伤模型小鼠肺组织白介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)表达的影响。方法 30只昆明小鼠随机分为正常对照组、急性肺损伤组和Wortmannin处理组。采用腹腔注射LPS(10 mg/kg)建立小鼠急性肺损伤模型,对照组腹腔注射同体积的生理盐水,Wortmannin处理组则于造模前2 h腹腔注射Wortmannin(1.4 mg/kg)。LPS注射后6 h处死大鼠,计算肺组织湿/干重(W/D)比值,Western blot方法检测三组小鼠肺组织内IL-1β和TNF-α蛋白的表达变化,RT-PCR方法检测三组小鼠肺组织内IL-1βmRNA和TNF-αmRNA的表达变化。结果急性肺损伤组小鼠肺组织IL-1β和TNF-α蛋白及mRNA表达水平显著上升,显著高于正常对照组(0.05);相比于急性肺损伤组小鼠,Wortmannin处理组小鼠肺组织IL-1β和TNF-α蛋白及mRNA表达水平显著降低(0.05)。结论 Wortmannin能抑制急性肺损伤小鼠肺组织IL-1β和TNF-α表达。     

IL-18在脓毒症肺损伤发生发展过程中的作用及机制研究北大核心CSCD

目的:研究IL-18在脓毒症肺损伤发展过程中的作用及调控机制。方法:成年雄性野生型C57BL/6(WT)和IL-18基因敲除(IL-18-/-)小鼠分为野生型小鼠对照组(WT组)、脂多糖(LPS)处理的野生型小鼠组(WT LPS组)、IL-18基因敲除的小鼠对照组(IL-18-/-组)、LPS处理的IL-18基因敲除小鼠组(IL-18-/-LPS组)。腹腔注射LPS(15 mg/kg)建立小鼠脓毒症模型,对照组注射等量生理盐水。观察各组小鼠72 h生存率并处死小鼠,HE染色观察肺部病理组织变化,RT-PCR及Western blot检测各组小鼠肺组织IL-18 mRNA及蛋白表达,免疫荧光检测肺组织IL-18表达及定位,TUNEL染色检测肺组织细胞凋亡,流式细胞术检测肺组织Treg/Th17比例,ELISA检测炎症因子TNF-α、IL-17A、TGF-1β、IL-10表达。Western blot检测各组小鼠肺组织RORγt、FoxP3蛋白表达及STAT3磷酸化水平。结果:LPS诱导后,小鼠肺组织高表达IL-18。与WT LPS组相比,IL-18-/-LPS组小鼠生存率显著提高,肺组织病理损伤减轻,凋亡细胞显著减少,Treg/Th17比例提高,抑炎因子TGF-1β、IL-10表达增加,而促炎因子TNF-α、IL-17A表达明显减少,肺组织RORγt蛋白表达增加,而STAT3磷酸化水平降低。结论:IL-18可通过上调STAT3磷酸化水平,促进Treg/Th17免疫失衡及炎症因子分泌,从而加剧脓毒症急性肺损伤。     

N-乙酰半胱氨酸通过抑制ROS-NLRP3信号通路诱导巨噬细胞的M2型分化改善脓毒症肺损伤

目的探讨N-乙酰半胱氨酸(NAC)对脓毒症肺损伤的作用及机制。方法 48只Balb/c小鼠随机分为对照组、模型组、NAC低剂量组、中剂量组、高剂量组和阳性药组(n=8)。给予LPS前1 h,腹腔注射药物干预研究。给予LPS 6 h后麻醉,收集肺泡灌洗液(BALF)和血清。苏木精-伊红染色检测肺损伤病理变化;ELISA检测BALF和血清中TNF-α、IL-6和IL-1β含量;Western blot检测组织中NLRP3、Caspase1和ASC蛋白水平;流式细胞术分析M2型巨噬细胞表型;商用试剂盒测量组织中MDA、MPO、SOD、GSH和ROS含量。结果与模型组比较,NAC组小鼠肺损伤显著缓解,且BALF和血清中TNF-α、IL-6和IL-1β含量减少。NAC组较模型组小鼠BALF中M2型巨噬细胞表型增强,且NLRP3、Caspase1和ASC蛋白下调。NAC组小鼠肺组织中MDA、MPO和ROS含量减少,同时,SOD和GSH含量增加。结论 N-乙酰半胱氨酸对急性肺损伤具有保护作用,且可能与抑制ROS-NLRP3信号通路诱导巨噬细胞的M2型分化有关。     

藤黄酸减轻脂多糖诱导的小鼠急性肺损伤

目的探讨藤黄酸(GA)对脂多糖(LPS)所致小鼠急性肺损伤的保护作用及其机制。方法采用尾静脉注射LPS(4 mg/kg)建立小鼠急性肺损伤模型。实验将小鼠随机分为对照组(control组)、模型组(model组)、藤黄酸组(GA组)和藤黄酸预处理组(GA+LPS组),6 h后测定肺湿/干重比值(W/D);检测髓过氧化物酶(MPO)活性;检测肺泡灌洗液(BALF)中蛋白含量和白细胞计数;ELISA检测肺匀浆中白介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)含量。结果模型组小鼠肺W/D、MPO活性、BALF中蛋白含量和白细胞数量均增加,肺组织IL-1β和TNF-α水平升高(均P0.01);藤黄酸预处理可减轻LPS引起的以上指标变化(均P0.05)。结论 GA可减轻LPS诱导的急性肺损伤,其机制可能与降低肺组织IL-1β和TNF-α的含量、抑制中性粒细胞在肺部的聚集和减轻肺部水肿相关。     

右美托咪啶减轻急性肺损伤的机制研究

目的探讨右美托咪啶(Dex)对小鼠急性肺损伤(ALI)的影响及潜在作用机制。方法将32只C57BL/6雄性小鼠随机分为空白对照组(Sham组)、右美托咪啶组(Dex组)、脂多糖组(LPS组)和药物干预组(LPS+Dex组)。LPS组和LPS+Dex组小鼠通过腹腔注射LPS(10 mg/kg)构建小鼠ALI模型,Dex组及LPS+Dex组小鼠腹腔注射Dex(40μg/kg),Sham组和LPS组注射等剂量生理盐水。LPS注射12 h后,采用HE染色比较各组小鼠肺损伤状况;qRT-PCR检测各组小鼠肺组织中促炎性细胞因子IL-1β、TNF-α、IL-6和MCP-1 mRNA的表达;Western blot检测各组小鼠肺组织中GPX4、COX2和转录因子红系2相关因子2(Nrf2)蛋白的表达。结果与Sham组相比,LPS组小鼠肺损伤评分明显升高(P0.05),促炎性细胞因子IL-1β、TNF-α、IL-6和MCP-1mRNA表达水平均明显升高(P0.05),肺组织铁死亡标志物GPX4蛋白表达水平明显降低(P0.05),COX2蛋白表达水平则明显升高(P0.05),肺组织中Nrf2蛋白表达水平明显降低(P0.05)。与LPS组相比,LPS+Dex组小鼠肺损伤评分明显降低(P0.05),IL-1β、TNF-α、IL-6和MCP-1 mRNA表达水平明显降低(P0.05),GPX4的蛋白表达水平明显升高(P0.05),COX2蛋白表达水平则明显降低(P0.05),Nrf2的蛋白表达水平也明显升高(P0.05)。结论 Dex可能通过激活Nrf2抑制小鼠肺组织铁死亡,进而发挥肺保护作用。     

钩藤碱降低内毒素血症小鼠死亡率的机制研究

目的： 观察钩藤碱对内毒素血症小鼠死亡率及器宫损伤的影响，并探讨其作用机制。方法: 雄性小鼠随机分为对照组、脂多糖（LPS）组、钩藤碱（Rhy）防治组和Rhy对照组，分别予以生理盐水、Rhy皮下注射，1 time/d，连续3 d，第3 d皮下注射后1 h，腹腔注射生理盐水或LPS（20 mg/kg）。观察各组小鼠的死亡率，肺、小肠组织病理改变；测定注射LPS后12 h各组肺湿/干重比值 (W/D)及血清丙氨酸氨基转移酶（ALT）、门冬氨酸氨基转移酶（AST）、尿素氮（BUN）和肌酐（Cr）的水平；用酶联免疫吸附法（ELISA）测定血清肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）及白细胞介素-10（IL-10）的含量，用硝酸还原酶法试剂盒测定血清一氧化氮（NO）的含量。进一步复制盲肠结扎穿孔的脓毒症模型， 观察钩藤碱对脓毒症小鼠生存率的影响。结果: LPS攻击后24 h小鼠的生存率明显低于对照组，8、16 mg/kg的钩藤碱防治组小鼠生存率高于LPS组。但钩藤碱并不能降低CLP小鼠的死亡率，而且，8 mg/kg钩藤碱治疗组小鼠的死亡率反而高于CLP小鼠的死亡率。LPS攻击后12 h病理检查发现LPS组小鼠肺及小肠组织均有严重的炎症表现；肺W/D、血清ALT、AST、BUN、Cr水平显著高于对照组；LPS攻击后2 h血清TNF-α、IL-1β及IL-10含量及8 h血清NO水平显著高于对照组。LPS攻击后12 h， Rhy防治组肺及小肠组织损伤无明显改善；肺W/D、血清ALT、AST、BUN、Cr水平与LPS组比较无显著差异；LPS攻击后2 h血清TNF-α水平显著低于LPS组，但2 h血清IL-1β及IL-10含量及8 h血清NO水平与LPS组比较无显著差异。结论: 钩藤碱能降低内毒素血症小鼠的死亡率，但不能降低脓毒症小鼠的死亡率，抑制TNF-α的生成可能是钩藤碱降低内毒素血症小鼠死亡率的机制之一。     

右美托咪定减轻脂多糖致大鼠急性肺损伤

目的探讨右美托咪定在减轻脂多糖(LPS)致大鼠急性肺损伤(ALI)中的作用及相关机制。方法将SD大鼠随机分为:对照组、脂多糖组(LPS组,腹腔注射LPS 5 mg/kg)和右美托咪定+脂多糖组(LD组,腹腔注射右美托咪定25μg/kg+LPS 5 mg/kg)。6 h后,收集左肺支气管肺泡灌洗液(BALF),ELISA检测BALF中白细胞介素-1β(IL-1β),白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的含量;取右肺组织进行HE染色观察肺组织病理学变化,并进行肺损伤评分;计算肺组织湿重/干重(W/D)的比值;Western blot检测肺组织中HGMB1、TLR4和TLR2蛋白的表达。结果与对照组相比,LPS组中肺损伤评分、W/D比值、BALF中IL-1β、IL-6、TNF-α的含量明显升高(P0.05),且肺组织中HMGB1,TLR2,TLR4的表达上调(P0.05);与LPS组相比,LD组中各指标变化显著减轻(P0.05)。结论右美托咪定能够减轻LPS致大鼠急性肺损伤,可能与HGMB1和TLRs蛋白表达的抑制有关。     

人参皂甙RB2 抑制NF-κB 的激活对LPS 诱导的新生小鼠急性肺损伤的免疫调节作用

目的:研究人参皂苷RB2对脂多糖(LPS)诱导的新生小鼠急性肺损伤的免疫功能的影响及相关分子机制。方法:将新生小鼠随机分为四组:健康组(Ctrl);人参皂苷单独处理组(GR2):健康小鼠腹腔注射GR2 50 mg/kg BW; LPS诱导组(LPS):腹腔注射0. 6 mg/kg BW的LPS,连续5 d; LPS+GR2组:模型小鼠腹腔注射人参皂苷50 mg/kg BW。HE染色观察肺组织损伤情况; TUNEL染色观察肺组织细胞凋亡; Western blot检测Caspase-3和Caspase-9的表达; ELISA检测炎症因子IL-6、IL-1β、TNF-α和IL-10的含量; Western blot检测高迁移率族蛋白B1(HMGB1)、巨噬细胞移动抑制因子(MIF)和NF-κBp 65的表达水平。结果:LPS组与对照组相比,肺组织出现明显的损伤,肺泡壁明显增厚,出现大量炎性细胞浸润;染色呈阳性的凋亡细胞比率显著提高; Caspase-3和Caspase-9的表达水平显著上调;炎症因子IL-6、IL-1β和TNF-α的含量显著上升,IL-10的含量没有明显变化; HMGB1和MIF的表达显著上调;磷酸化的NF-κB p65的表达显著上调。LPS+GR2组与LPS组相比,肺组织损伤明显得到缓解,多数肺泡结构完整,仅有少量炎性细胞浸润;凋亡细胞的比率显著下降; Caspase-3和Caspase-9的表达水平显著下调;炎症因子IL-6、IL-1β和TNF-α的含量显著下降,IL-10的含量没有明显的变化; HMGB1和MIF的表达显著下调;磷酸化的NF-κB p65的表达显著下调。结论:人参皂苷RB2抑制NF-κB的激活,进而调节LPS诱导的新生小鼠急性肺损伤的免疫反应。     

丙酮酸乙酯对LPS致急性肺损伤大鼠肺组织NF-κB p65及TNF-α和IL-lβ的影响

目的:观察丙酮酸乙酯(EP)对内毒素性急性肺损伤(ALI)大鼠肺组织核转录因子-κB(NF-κB)p65表达及肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)含量的影响,探讨EP可能的肺保护机制。方法:雄性SD大鼠30只随机分为三组(n均=10):正常对照组,静脉注射与其它二组等量生理盐水;LPS组,静脉注射脂多糖(LPS)5mg/kg复制大鼠ALI模型;EP+LPS组,于静脉注射LPS前1h腹腔内注射EP(40mg/kg)。所有动物于注射LPS或生理盐水后4h颈动脉放血处死,取肺组织用Westernblot测定其NF-κBp65的表达,用ELISA测定其TNF-α和IL-1β的含量。结果:与对照组相比,LPS组、EP+LPS组肺组织NF-κBp65表达增加,肺组织TNF-α和IL-lβ含量升高(P0.05);与LPS组相比,EP+LPS组肺组织NF-κBp65表达降低,肺组织TNF-α和IL-lβ含量降低(P0.05)。结论:EP通过下调大鼠LPS诱导的肺组织NF-κBp65表达,降低了TNF-α和IL-lβ的释放。EP可减轻ALI大鼠肺的炎症反应。     

microRNA-1285-3p在脓毒症急性肺损伤中的表达及其与炎症因子水平的相关性

目的探讨microRNA(miRNA)-1285-3p在脓毒症急性肺损伤(ALI)中的表达及其与炎症因子、肿瘤坏死因子(TNF)-α、白介素(IL)-6水平的相关性。方法对大鼠进行腹腔注射内毒素脂多糖(LPS)诱导急性肺损伤大鼠模型,选取相同数量的大鼠作为对照组;各模型组大鼠注射LPS 4 h、8 h、12 h以及24 h后,观察各组大鼠肺组织病理学变化,并检测肺组织中microRNA-1285-3p、TNF-α与IL-6的含量变化。利用LPS刺激大鼠肺泡巨噬细胞(NR8383细胞),在4 h、8 h、12 h以及24 h不同时间点检测细胞中microRNA-1285-3p、TNF-α与IL-6的含量变化,并分析细胞中microRNA-1285-3p的表达与TNF-α、IL-6表达的相关性。以生物信息学方法对microRNA-1285-3p及其靶基因进行预测,上调或下调NR8383细胞microRNA-1285-3p表达,以LPS刺激后,12 h后观察microRNA-1285-3p的表达变化,利用蛋白免疫印迹试验(Western blot)检测24 h后细胞中microRNA-1285-3p靶蛋白与NF-κB的表达变化。结果 LPS 4 h组、LPS 8 h组、LPS 12 h组、LPS 24 h组肺组织及NR8383细胞系中的microRNA-1285-3p表达量均分别低于对照组(P0.05),而TNF-α、IL-6水平均高于对照组(P0.05);microRNA-1285-3p表达与TNF-α、IL-6均呈负相关关系(均P0.05);microRNA-1285-3p mimic组Notch1蛋白(Notch信号通路受体蛋白之一)的表达低于对照组(P0.05);microRNA-1285-3p抑制剂(inhibitor)组Notch1蛋白的表达水平高于对照组(P0.05)。结论microRNA-1285-3p可能通过调控脓毒症急性肺损伤大鼠的Notch1蛋白的表达,影响炎症因子TNF-α与IL-6的表达,参与其免疫炎症调控过程。     

Cavidine Ameliorates Lipopolysaccharide-Induced Acute Lung Injury <Emphasis Type="Italic">via</Emphasis> NF-κB Signaling Pathway <Emphasis Type="Italic">in vivo</Emphasis> and <Emphasis Type="Italic">in vitro</Emphasis>

Acute lung injury (ALI) is characterized by widespread inflammation in the lungs and alveolar-capillary destruction, causing high morbidity and mortality. Cavidine, isolated from Corydalis impatiens, have been exhibited to have potent anti-inflammatory effects in previous studies. The purpose of this study was to evaluate the protective effect of cavidine on lipopolysaccharide (LPS)-induced ALI and to enunciate the underlying in vivo and in vitro mechanisms. Mice were intraperitoneally administrated with cavidine (1, 3, or 10 mg/kg) at 1 and 12 h, prior to the induction of ALI by intranasal administration of LPS (30 mg/kg). Blood samples, lung tissues, and bronchoalveolar lavage fluid (BALF) were harvested after LPS challenge. Furthermore, we used LPS-induced lung epithelial cells A549 to examine the mechanism of cavidine to lung injury. The results showed that pretreatment with cavidine significantly decreased lung wet-to-dry weight (W/D) ratio, reduced pro-inflammatory cytokine levels including TNF-α and IL-6 in BALF and serum from LPS-stimulated mice, and attenuated lung histopathological changes. In addition, western blot results showed that cavidine inhibited the phosphorylation of nuclear factor-kappaB (NF-κB) p65 and IκBα induced by LPS. In conclusion, our results demonstrate that cavidine protects against LPS-induced acute lung injury in mice via inhibiting of pro-inflammatory cytokine TNF-α and IL-6 production and NF-κB signaling pathway activation. Taken together, cavidine may be useful for the prevention and treatment of pulmonary inflammatory diseases, such as ALI.     

Treatment of Low Molecular Weight Heparin Inhibits Systemic Inflammation and Prevents Endotoxin-Induced Acute Lung Injury in Rats

To determine whether low molecular weight heparin (LMWH) is able to reduce pulmonary inflammation and improve the survival in rats with endotoxin-induced acute lung injury (ALI). Rat ALI model was reproduced by injection of lipopolysaccharide (LPS) into tail vein. Rats were divided randomly into three groups: control group, ALI group, LMWH-treated group. Blood was collected and lung tissue was harvested at the designated time points for analysis. The lung specimens were harvested for morphological studies, streptavidin-peroxidase immunohistochemistry examination. Lung tissue edema was evaluated by tissue water content. The levels of lung tissue myeloperoxidase (MPO) were determined. Meanwhile, the nuclear factor-kappa B (NF-κB) activation, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) levels and high mobility group box 1 (HMGB1) and intercellular adhesion molecule-1 (ICAM-1) protein levels in the lung were studied. In survival studies, a separate group of rats were treated with LMWH or sterile saline after LPS administration. Then, the mortality was recorded. Treatment with LMWH after ALI was associated with a reduction in the severity of LPS-induced lung injury. Treatment with LMWH significantly decreased the expression of TNF-α, IL-1β, HMGB1 and ICAM-1 in the lung of ALI rats. Similarly, treatment with LMWH dramatically diminished LPS-induced neutrophil sequestration and markedly reduced the enhanced lung permeability. In the present study, LMWH administration inhibited the nuclear translocation of NF-κB in the lung. Survival was significantly higher among the LMWH-treated group compared with the ALI group. These data suggest that LMWH attenuates inflammation and prevents lethality in endotoxemic rats.     

Anti-inflammatory Role of Pilose Antler Peptide in LPS-Induced Lung Injury

The present study was designed to investigate the effects of pilose antler peptide (PAP) on lipopolysaccharide (LPS)-induced lung injury. BalB/c mice intraperitoneally received PAP (10 and 20 mg/kg) or dexamethasone (2 mg/kg) 1 h prior to intratracheal instillation of LPS. PAP significantly decreased lung wet-to-dry weight (W/D) ratio and lung myeloperoxidase (MPO) activity and restored LPS-induced lung histopathological changes. PAP also increased super oxide dismutase (SOD) level and inhibited malondialdehyde (MDA) content and levels of pro-inflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in bronchoalveolar lavage fluid (BALF) in LPS-stimulated mice. Furthermore, we demonstrated that PAP inhibited Rho/NF-κB pathway in LPS-induced mice. Our experimental results indicated that the protective mechanism of PAP might be attributed partly to the inhibition of Rho/NF-κB pathway.     

Inhibition of PKR ameliorates lipopolysaccharide-induced acute lung injury by suppressing NF-κB pathway in mice

Acute lung injury (ALI) is characterized by dramatic lung inflammation and alveolar epithelial cell death. Although protein kinase R (PKR) (double-stranded RNA-activated serine/threonine kinase) has been implicated in inflammatory response to bacterial cell wall components, whether it plays roles in lipopolysaccharide (LPS)-induced ALI remains unclear. This study was aimed to reveal whether and how PKR was involved in LPS-induced ALI pathology and the potential effects of its specific inhibitor, C16 (C₁₃H₈N₄OS). During the experiment, mice received C16 (100 or 500 ug/kg) intraperitoneally 1?h before intratracheal LPS instillation. Then, whole lung lavage was collected for analysis of total protein levels and proinflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and IL-6. The lungs were tested for Western blot, transferase-mediated dUTP nick-end labeling (TUNEL) stain and immunohistochemistry. Results showed that PKR phosphorylation increased significantly after LPS instillation. Furthermore, PKR specific inhibition attenuated LPS-induced lung injury (hematoxylin and eosin stain), reduced lung protein permeability (total protein levels in whole lung lavage) and suppressed proinflammatory cytokines (TNF-α, IL-1β and IL-6) and lung apoptosis (TUNEL stain and caspase3 activation). Moreover, mechanism-study showed that C16 significantly suppressed I kappa B kinase (IKK)/I kappa B alpha (IκBα)/NF-κB signaling pathway after LPS challenge. These findings suggested that PKR inhibition ameliorated LPS-induced lung inflammation and apoptosis in mice by suppressing NF-κB signaling pathway.     

The Effect of Epigallocatechin Gallate on Lipopolysaccharide-Induced Acute Lung Injury in a Murine Model

This study was performed to evaluate the effects of epigallocatechin 3 gallate (EGCG) on lipopolysaccharide (LPS)-induced acute lung injury in a murine model. In the present study, production of TNF-α and MIP-2 and activation of extracellular signal-regulated kinases (ERK)1/2, c-Jun amino terminal kinases (JNK) and p38 in RAW264.7 cells were measured. EGCG inhibited the production of TNF-α and MIP-2, and attenuated phosphorylation levels of ERK1/2 and JNK, but not p38 in RAW264.7 cells stimulated with LPS. Also, EGCG attenuated the production of TNF-α and MIP-2, and the phosphorylation of ERK1/2 and JNK in the lungs of mice administered with LPS intratracheally. It reduced wet/dry weight ratio, histological severities, and neutrophil accumulation in the lungs in mice given LPS. Our results showed that EGCG attenuated LPS-induced lung injury by suppression of the MIP-2 and TNF-α production, and ERK1/2 and JNK activation in macrophage stimulated with LPS.     

罗哌卡因通过 HMGB1 / NF-κB 通路减轻 LPS 诱导的小鼠急性 肺损伤

目的 探讨罗哌卡因 ( ropivacaine, Rop) 对脂多糖 ( lipopolysaccharide, LPS) 诱导的小鼠急性 肺损伤 (acute lung injury, ALI) 的作用及其机制。 方法 气管内滴注 LPS 诱导肺损伤小鼠模型, 并将小鼠 随机分为 6 组: 对照组、 LPS 组、 罗哌卡因 0. 25、 0. 5、 1 μmol / L 组和右美托咪定 (dexmedetomidine, Dex) 100 μg / kg 组。 Hematoxylin-eosin (H&E) 染色评估肺组织的组织病理学变化; ELISA 法测定肺组织中髓过 氧化物酶 (myeloperoxidase, MPO)、 丙二醛 ( malondialdehyde, MDA)、 超氧化物歧化酶 ( superoxide dismutase, SOD) 和谷胱甘肽过氧化物酶 ( glutathione peroxidase, GSH-Px) 的活性; 检测血清中 IL-6、 IL-1β 和肿瘤坏死因子-α (tumor necrosis factor-α, TNF-α) 的表达; Western 印迹检测 HMGB1 / NF-κB 通路相关蛋 白的表达。 结果 与对照组比较, LPS 诱导肺泡外膜增厚、 出血和肺水肿; 肺损伤评分和肺含水量增加; MPO 和 MDA 活性增加, SOD 和 GSH-Px 水平降低; IL-6、 IL-1β 和 TNF-α 水平升高; HMGB1 蛋白和 NF-κB P65 磷酸化水平升高, 有显著性差异 (P< 0. 05)。 与 LPS 组比较, 罗哌卡因 0. 5、 1 μmol / L 组小鼠肺损伤 程度明显减轻; MPO 和 MDA 活性降低, SOD 和 GSH-Px 水平升高; IL-6、 IL-1β 和 TNF-α 水平降低; HMGB1 蛋白和 NF-κB P65 磷酸化水平降低, 有显著性差异 (P< 0. 05)。 结论 罗哌卡因通过抑制 HMGB1 / NF-κB 通路, 有效减弱了 LPS 引起的肺组织损伤。     

Protective effect of gossypol on lipopolysaccharide-induced acute lung injury in mice

Objective

Gossypol has been reported to have anti-inflammatory properties. The purpose of this study was to evaluate the effect of gossypol on acute lung injury (ALI) induced by lipopolysaccharide (LPS) in mice.

Methods

Male BALB/c mice were pretreated with gossypol 1 h before intranasal instillation of LPS. Then, 7 h after LPS administration, the myeloperoxidase in histology of lungs, lung wet/dry ratio and inflammatory cells in the bronchoalveolar lavage fluid (BALF) were determined. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) in the BALF were measured by ELISA. The extent of phosphorylation of IκB-α, p65 NF-κB, p46–p54 JNK, p42–p44 ERK, and p38 were detected by western blot.

Results

Gossypol markedly attenuated the LPS-induced histological alterations in the lung and inhibited the production of TNF-α, IL-1β and IL-6. Additionally, gossypol reduced the inflammatory cells in BALF, decreased the wet/dry ratio of lungs and inhibited the phosphorylation of IκB-α, p65 NF-κB, p46–p54 JNK, p42–p44 ERK, and p38 caused by LPS.

Conclusion

The data suggest that anti-inflammatory effects of gossypol against the LPS-induced ALI may be due to its ability of inhibition of the NF-κB and MAPKs signaling pathways. Gossypol may be a promising potential therapeutic reagent for ALI treatment.     

Chikusetsusaponin V attenuates lipopolysaccharide-induced liver injury in mice

Chikusetsusaponin V (CsV), a saponin from Panax japonicus, has been reported to inhibit inflammatory responses in lipopolysaccharide (LPS)-induced macrophage cells. However, whether CsV could alleviate LPS-induced liver injury in vivo and the potential mechanisms involved remain unclear. In the present study, we investigated the anti-inflammatory effects of CsV on LPS-induced acute liver injury in mice and further explored the potential mechanisms involved. Our results showed that CsV significantly attenuated elevation of alanine transaminase (ALT) and aspartate aminotransferase (AST) levels and improved liver histopathological changes in LPS-induced mice. In addition, CsV decreased serum tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) levels and inhibited mRNA expressions of inducible nitric oxide synthase (iNOS), TNF-α and IL-1β in LPS challenged mice. Furthermore, CsV inhibited nuclear factor kappa B (NF-κB) activation by downregulating phosphorylated NF-κB, IκB-α, ERK, c-Jun N-terminal kinase (JNK) and p38 levels in the liver tissue, which ultimately decreased nucleus NF-κB protein level. In conclusion, our data suggested that CsV could be a promising drug for preventing LPS challenged liver injury since it attenuated LPS-induced inflammatory responses, partly via inhibiting NF-κB and MAPK signaling pathways.     

Cepharanthine, an Alkaloid from Stephania cepharantha Hayata, Inhibits the Inflammatory Response in the RAW264.7 Cell and Mouse Models

The aim of this study was to investigate the protective effects of cepharanthine (CEP) on inflammation in lipopolysaccharide (LPS)-stimulated RAW264.7 cells in vitro and a LPS-induced lung injury model in vivo. RAW264.7 cells were treated with various concentrations of CEP for 1 h followed by incubation with or without 1 μg/ml LPS for 18 h. TNF-α, IL-6, and IL-1β in the supernatants were measured by ELISA. Nuclear factor-κB (NF-κB) and mitogen-activated protein kinase pathways were analyzed by Western blot. Mice were randomly divided into control group, LPS group, CEP?+?LPS group, and dexamethasone?+?LPS group. A male BALB/c mouse model of acute lung injury was induced by LPS. Bronchoalveolar lavage fluid was collected for inflammatory cell count and cytokine assays. Histopathologic examination was performed on mice that were not subjected to bronchoalveolar lavage fluid collection. CEP dose-dependently inhibited the release of TNF-α, IL-6, and IL-1β in LPS-stimulated RAW264.7 cells. Significantly, CEP dose-dependently suppressed NF-κB activation, IκBα degradation, and phosphorylation of ERK, JNK, and p38 induced by LPS. In vivo, it was also observed that CEP attenuated lung histopathologic changes and down-regulated the level of pro-inflammatory cytokines, including TNF-α, IL-1β, and IL-6, in the mouse acute lung injury model. These results suggest that CEP potentially decreases inflammation in vitro and in vivo and might be a therapeutic agent against inflammatory diseases.