肝素的抗炎作用与抗细胞粘附调节

肝素类药物具有抗凝以外的包括抗炎在内的多种生物学活性。炎症反应是多种因子、细胞参与的复杂的病生理过程,其物质基础是粘附分子介导的白细胞粘附及其粘附级联反应。近来研究证实肝素抗炎机制主要与抗细胞粘附调节有关,肝素通过竞争抑制L—、P—选择素与其配基sLe^X的结合,阻止白细胞粘附活化及调抑炎症级联反应,进而起到抗炎作用。对肝素抗炎机制的深入研究,将有助于进一步阐明抗粘附／抗炎的作用机制。     

H_2S与急性肺损伤研究进展

<正>急性肺损伤是一种过度炎症反应,它是在感染、创伤等过程中导致的一种过度炎性反应,严重时则被称为急性呼吸窘迫综合征。多年来,急性肺损伤一直是临床常见的严重疾病,急性肺损伤的发病机制也未完全阐明。大量研究证明,继NO和CO之后的第3种信号分子H2S是重要的信使分子,并具有血管功能调节的作用。还有研究证明,H2S具有多方面的生物学作用,如参与低氧性肺动脉高压和感染性休克以及减轻兴奋性氨基酸对神经元的毒性作用等。内源性     

抗炎多肽AF-2对LPS诱导的小鼠急性肺损伤的保护

目的研究抗炎多肽AF-2（antiflammin-2）对内毒素（LPS）诱导的小鼠急性肺损伤的保护作用。方法Balb/c雄性小鼠37只,随机分为3组,对照组（n=10）、急性肺损伤（ALI）模型组（n=14）和AF-2治疗组（n=13）,模型组和治疗组腹腔注射大肠杆菌内毒素复制小鼠肺损伤模型,治疗组同时注射抗炎多肽AF-2,对照组和模型组注射等量的生理盐水。在0h、6h和12h记录动物的呼吸频率,12h处死动物,肺组织切片观察肺病理变化,ELISA法检测血清细胞因子。结果6h和12hAF-2治疗组动物呼吸频率均低于模型组。肺组织病理显示AF-2对LPS诱导的小鼠ALI肺组织的渗出、炎细胞浸润有一定的抑制作用。AF-2治疗组与ALI模型组比较血清IL-6水平明显下降。结论AF-2对内毒素诱导的小鼠急性肺损伤有一定的保护作用。     

抗氧化治疗与急性肺损伤

急性肺损伤（acute lung injury,ALI）是指心源性以外的各种肺内外致病因素导致的急性、进行性呼吸衰竭,其主要的病理特征是肺微血管通透性增高而导致的肺泡渗出液中富含蛋白质的肺水肿及透明膜形成,可伴有肺间质纤维化。病理生理改变以肺顺应性降低、     

硫化氢在不同诱因致急性肺损伤中的作用

硫化氢（Hydrogensulfide,H2S）是一种具有臭鸡蛋气味的有毒气体,其毒理作用的主要靶器是中枢神经系统和呼吸系统,亦可伴有心脏等多器官损害。上世纪90年代中期,人们发现体内含硫氨基酸代谢生成的内源性H2S对神经系统特别是海马的功能具有调节作用,并可调节消化道和血管平滑肌的张力,从而对H2S有了新认识。如今,     

粘附蛋白与内皮细胞

RGD是粘附蛋白与细胞接合的活性部分之一。血小板精白蛋Ⅱb/Ⅲa是识别RGD的受体家族,可以与RGD结合。纤维蛋白(原)能特异地刺激内皮细胞合成和分泌PGI2及t-PA。     

双歧杆菌粘附大肠癌细胞相关因素探讨

本文以青春双歧杆菌ＤＭ９２２７粘附大肠癌细胞ｃｌ－１８７为实验模型,探讨粘附的相关因素。结果表明：粘附需要一定的菌浓度（＞１０６ＣＦＵ／ｍｌ）,一定的温度（３７℃）及一定的时间（＞１０ｈ）,且偏酸更有利于粘附。破坏菌体表面的糖结构使粘附上升,而破坏蛋白结构则使粘附下降。     

急性肺损伤保护作用机制研究进展

急性肺损伤(acute lung injury,ALI)是临床常见危重症,病死率极高,其发病机制极为复杂,临床上针对急性肺损伤缺乏有效的治疗药物及治疗手段。现从抗炎、抗氧化、内皮细胞保护及改善微循环等方面对急性肺损伤保护作用机制作一综述。     

人参皂苷Rb1对H9N2流感病毒诱导急性肺损伤小鼠抗氧化酶的影响

目的探讨人参皂苷Rb1(G-Rb1)对肺损伤小鼠抗氧化酶活力的影响。方法将195只6~8周龄BALB/c小鼠随机分为对照组、肺损伤模型组(ALI组)、人参皂苷Rb1组(G-Rb1组),每组65只。ALI与G-Rb1组采用100μL SI A/swine/HeBei/012/2008/猪流感病毒(H9N2 SIV)经鼻腔接种建立急性肺损伤模型,同时G-Rb1组腹腔注射人参皂苷Rb1液0.1 mL,剂量为10 mg/(kg·bw),连续7d;对照组鼻腔接种相同剂量生理盐水稀释的正常鸡胚尿囊液。观察临床症状、肺病理组织学变化,计算小鼠肺湿干重比、肺系数,检测小鼠肺组织T-SOD、MPO、CAT、GSH-PX活力。结果从第2天末开始ALI组大部分小鼠出现高度的精神沉郁,呼吸极度困难,采食量明显减少,体重下降。肺部明显水肿、淤血和出血,炎性细胞渗出,对照组小鼠各器官未见异常。肺系数及肺干湿重比逐渐升高,第8天开始下降,第14天趋于正常。G-Rb1组症状明显轻于攻毒组,症状出现较缓,症状较轻,死亡时间延迟,死亡率降低。在第4、6、8天,与对照组比G-Rb1组和ALI组T-SOD及CAT活力显著降低(P0.01),组间比,G-Rb1组明显高于ALI组(P0.05);在各时间点,与对照组比,ALI组GSH-PX活力显著降低(P0.01),而GRb1组则显著升高(P0.01),实验组组间差异显著(P0.01)。结论 G-Rb1在一定浓度范围内,具有提高小鼠抗氧化酶活力作用,一定程度上改善H9N2猪流感病毒对肺组织的氧化损伤。     

乌司他丁对急性肺损伤/急性呼吸窘迫综合征的治疗研究

目的研究大剂量乌司他丁在急性肺损伤/急性呼吸窘迫综合征中的治疗效果。方法回顾性分析2006年1月至2010年1月广西医科大学第一附属医院ICU收治的154例ALI/ARDS患者的临床资料,根据治疗方案分为乌司他丁组(UTI组)(n=80),对照组(n=74)。记录两组患者开始治疗、治疗第3天、治疗第7天的生命体征、动脉血气分析、血生化检查结果;记录患者在ICU治疗的转归。应用SPSS 13.0软件对结果进行统计学分析。结果经治疗3天UTI组呼吸频率低于对照组;动脉血气分析提示两组患者PaO2、PaO2/Fi O2、SaO2均有上升,UTI组PaO2/Fi O2略低于对照组(P0.01),而两组患者PaO2、SaO2比较无统计学差异。UTI组与对照组的死亡率比较(UTI组52.5%,对照组52.7%,P=0.980)无统计学差异,机械通气时间UTI组低于对照组[UTI组(14.8±3.9)天,对照组(16.7±4.2)天,P=0.020]。根据ALI/ARDS发生的病因分为肺内源性及肺外源性进行亚组分析(A组:肺内源性ALI/ARDS,使用UTI治疗;B组:肺内源性ALI/ARDS,不使用UTI治疗;C组:肺外源性ALI/ARDS,使用UTI治疗;D组:肺外源性ALI/ARDS,不使用UTI治疗),发现乌司他丁对肺外源性ALI/ARDS患者(C组)的ICU时间、ICU内死亡率及机械通气时间均低于不使用UTI的患者(D组)。结论大剂量乌司他丁用于ALI/ARDS的临床治疗可有效改善患者氧合指数,减少机械通气时间,且高血糖的发生率低,尤其是乌司他丁治疗肺外源性ALI/ARDS患者的预后优于肺内源性的ALI/ARDS。     

Effect of emodin on long non-coding RNA-mRNA networks in rats with severe acute pancreatitis-induced acute lung injury

Long non-coding RNAs (lncRNAs) contribute to disease pathogenesis and drug treatment effects. Both emodin and dexamethasone (DEX) have been used for treating severe acute pancreatitis-associated acute lung injury (SAP-ALI). However, lncRNA regulation networks related to SAP-ALI pathogenesis and drug treatment are unreported. In this study, lncRNAs and mRNAs in the lung tissue of SAP-ALI and control rats, with or without drug treatment (emodin or DEX), were assessed by RNA sequencing. Results showed both emodin and DEX were therapeutic for SAP-ALI and that mRNA and lncRNA levels differed between untreated and treated SAP-ALI rats. Gene expression profile relationships for emodin-treated and control rats were higher than DEX-treated and -untreated animals. By comparison of control and SAP-ALI animals, more up-regulated than down-regulated mRNAs and lncRNAs were observed with emodin treatment. For DEX treatment, more down-regulated than up-regulated mRNAs and lncRNAs were observed. Functional analysis demonstrated both up-regulated mRNA and co-expressed genes with up-regulated lncRNAs were enriched in inflammatory and immune response pathways. Further, emodin-associated lncRNAs and mRNAs co-expressed modules were different from those associated with DEX. Quantitative polymerase chain reaction demonstrates selected lncRNA and mRNA co-expressed modules were different in the lung tissue of emodin- and DEX-treated rats. Also, emodin had different effects compared with DEX on co-expression network of lncRNAs Rn60_7_1164.1 and AABR07062477.2 for the blue lncRNA module and Nrp1 for the green mRNA module. In conclusion, this study provides evidence that emodin may be a suitable alternative or complementary medicine for treating SAP-ALI.     

预防SARS病毒核酸疫苗的构建

本研究通过反转录-PCR获得了SARS冠状病毒辐条样蛋白、核衣壳蛋白和膜蛋白基因,将所获得的可能与免疫保护相关的基因克隆至核酸疫苗表达载体pcDNA-ThyA中,酶切及序列分析结果均表明载体构建正确,该候选DNA疫苗已用于动物免疫实验。     

Anti-inflammatory effects of novel curcumin analogs in experimental acute lung injury

Background

Acute lung injury (ALI) and its most severe form acute respiratory distress syndrome (ARDS) have been the leading cause of morbidity and mortality in intensive care units (ICU). Currently, there is no effective pharmacological treatment for acute lung injury. Curcumin, extracted from turmeric, exhibits broad anti-inflammatory properties through down-regulating inflammatory cytokines. However, the instability of curcumin limits its clinical application.

Methods

A series of new curcumin analogs were synthesized and screened for their inhibitory effects on the production of TNF-α and IL-6 in mouse peritoneal macrophages by ELISA. The evaluation of stability and mechanism of active compounds was determined using UV-assay and Western Blot, respectively. In vivo, SD rats were pretreatment with c26 for seven days and then intratracheally injected with LPS to induce ALI. Pulmonary edema, protein concentration in BALF, injury of lung tissue, inflammatory cytokines in serum and BALF, inflammatory cell infiltration, inflammatory cytokines mRNA expression, and MAPKs phosphorylation were analyzed. We also measured the inflammatory gene expression in human pulmonary epithelial cells.

Results

In the study, we synthesized 30 curcumin analogs. The bioscreeening assay showed that most compounds inhibited LPS-induced production of TNF-α and IL-6. The active compounds, a17, a18, c9 and c26, exhibited their anti-inflammatory activity in a dose-dependent manner and exhibited greater stability than curcumin in vitro. Furthermore, the active compound c26 dose-dependently inhibited ERK phosphorylation. In vivo, LPS significantly increased protein concentration and number of inflammatory cells in BALF, pulmonary edema, pathological changes of lung tissue, inflammatory cytokines in serum and BALF, macrophage infiltration, inflammatory gene expression, and MAPKs phosphorylation . However, pretreatment with c26 attenuated the LPS induced increase through ERK pathway in vivo. Meanwhile, compound c26 reduced the LPS-induced inflammatory gene expression in human pulmonary epithelial cells.

Conclusions

These results suggest that the novel curcumin analog c26 has remarkable protective effects on LPS-induced ALI in rat. These effects may be related to its ability to suppress production of inflammatory cytokines through ERK pathway. Compound c26, with improved chemical stability and bioactivity, may have the potential to be further developed into an anti-inflammatory candidate for the prevention and treatment of ALI.

Electronic supplementary material

The online version of this article (doi:10.1186/s12931-015-0199-1) contains supplementary material, which is available to authorized users.     

间充质干细胞移植治疗急性肺损伤的研究进展

急性肺损伤是一种临床常见的危重病症,临床上传统的治疗方法一般以尽早去除诱因、控制感染、机械通气及器官功能支持治疗为主。间充质干细胞属于成体干细胞的一种,能主动归巢至肺损伤部位,并通过向肺泡和支气管上皮细胞分化参与组织修复,同时间充质干细胞能够调节急性肺损伤时局部和全身炎症反应和免疫紊乱,从而发挥治疗作用,可能是治疗急性肺损伤的一个很有前景的方法。作者就间充质干细胞移植治疗急性肺损伤的研究进展进行综述。     

Anti-inflammatory effects of anethole in lipopolysaccharide-induced acute lung injury in mice

Aims

Anethole, the major component of the essential oil of star anise, has been reported to have antioxidant, antibacterial, antifungal, anti-inflammatory, and anesthetic properties. In this study, we investigated the anti-inflammatory effects of anethole in a mouse model of acute lung injury induced by lipopolysaccharide (LPS).

Main methods

BALB/C mice were intraperitoneally administered anethole (62.5, 125, 250, or 500 mg/kg) 1 h before intratracheal treatment with LPS (1.5 mg/kg) and sacrificed after 4 h. The anti-inflammatory effects of anethole were assessed by measuring total protein and cell levels and inflammatory mediator production and by histological evaluation and Western blot analysis.

Key findings

LPS significantly increased total protein levels; numbers of total cells, including macrophages and neutrophils; and the production of inflammatory mediators such as matrix metalloproteinase 9 (MMP-9), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and nitric oxide (NO) in bronchoalveolar lavage fluid. Anethole (250 mg/kg) decreased total protein concentrations; numbers of inflammatory cells, including neutrophils and macrophages; and the inflammatory mediators MMP-9, TNF-α and NO. In addition, pretreatment with anethole decreased LPS-induced histopathological changes. The anti-inflammatory mechanism of anethole in LPS-induced acute lung injury was assessed by investigating the effects of anethole on NF-κB activation. Anethole suppressed the activation of NF-κB by blocking IκB-α degradation.

Significance

These results, showing that anethole prevents LPS-induced acute lung inflammation in mice, suggest that anethole may be therapeutically effective in inflammatory conditions in humans.     

复方清下汤对脓毒症大鼠细胞因子调控及其肺损伤的作用

目的盲肠结扎穿孔导致大肠埃希菌腹膜炎进而建立脓毒症肺损伤大鼠模型,检测炎性反应时,细胞因子的调控变化,探讨肺水肿的形成机制。经复方清下汤处理后检测上述变化,以期为脓毒症肺损伤的防治提出可能的新途径。方法将健康SD大鼠随机分为4组,每组10只：假手术组（SHAM组）,只翻动盲肠,不做其他处理;脓毒症肺损伤组（模型组）,盲肠结扎穿孔诱发AL（急性肺损伤）I模型;盲肠结扎穿孔＋复方清下汤组（造模后立即灌胃给药,造模后8 h再次灌胃1次,剂量为10 m l/kg）;盲肠结扎穿孔＋头孢哌酮/舒巴坦组（抗生素舒普深）（造模后立即静脉注射1次,造模后8 h再次静脉注射1次,剂量为0.2 g/kg）造模24 h后收集标本。分别观察大鼠的一般状态,肺组织匀浆MPO的测定,留取下腔静脉血清进行TNF-α的测定。镜下观察肺组织病理形态学改变,测量肺湿/干比值的变化。结果与SHAM组比较,模型组MPO、TNF-α水平明显升高（P〈0.01）,肺间质和肺泡内水肿,伴大量红细胞渗出（出血）和纤维素沉积,肺泡间隔毛细血管内皮细胞高度肿胀。肺湿/干比值明显增加（P〈0.01）,抗生素及中药处理组与模型组比较,MPO、TNF-α水平明显降低（P〈0.01）,肺湿/干比值明显降低（P〈0.01）,肺组织镜下表现：中药处理组及抗生素组较模型组肺泡间隔变窄,毛细血管内皮细胞肿胀减轻,出血减轻,纤维素渗出明显减少。结论脓毒症大鼠肺损伤时细胞因子TNF-α过度表达,炎性介质的过度表达可能是造成脓毒症肺损伤的重要原因,而复方清下汤可以减轻脓毒症时的肺损伤和抑制TNF-α的表达,它们之间可能存在一定的联系。     

Regulation of Na,K-ATPase during acute lung injury

A hallmark of acute lung injury is the accumulation of a protein rich edema which impairs gas exchange and leads to hypoxemia. The resolution of lung edema is effected by active sodium transport, mostly contributed by apical Na⁺ channels and the basolateral located Na,K-ATPase. It has been reported that the decrease of Na,K-ATPase function seen during lung injury is due to its endocytosis from the cell plasma membrane into intracellular pools. In alveolar epithelial cells exposed to severe hypoxia, we have reported that increased production of mitochondrial reactive oxygen species leads to Na,K-ATPase endocytosis and degradation. We found that this regulated process follows what is referred as the Phosphorylation–Ubiquitination–Recognition–Endocytosis–Degradation (PURED) pathway. Cells exposed to hypoxia generate reactive oxygen species which activate PKCζ which in turn phosphorylates the Na,K-ATPase at the Ser18 residue in the N-terminus of the α1-subunit leading the ubiquitination of any of the four lysines (K16, K17, K19, K20) adjacent to the Ser18 residue. This process promotes the α1-subunit recognition by the μ2 subunit of the adaptor protein-2 and its endocytosis trough a clathrin dependent mechanism. Finally, the ubiquitinated Na,K-ATPase undergoes degradation via a lysosome/proteasome dependent mechanism.     

Silencing of Paralemmin-3 Protects Mice from lipopolysaccharide-induced acute lung injury

Excessive inflammatory response induced by lipopolysaccharide (LPS) plays a critical role in the development of acute lung injury (ALI). Paralemmin-3 (PALM3) is a novel protein that can modulate LPS-stimulated inflammatory responses in alveolar epithelial A549 cells. However, it remains unclear whether it is involved in the progression of ALI in vivo. Therefore, we studied the role of PALM3 in the pathogenesis of ALI induced by LPS. ALI was induced by LPS peritoneal injection in C57BL/6J mice. Lentivirus-mediated small interfering RNA (siRNA) targeting the mouse PALM3 gene and a negative control siRNA were intranasally administered to the mice. We found that the expression of PALM3 was up-regulated in the lung tissues obtained from the mouse model of LPS-induced ALI. The LPS-evoked inflammatory response (neutrophils and the concentrations of proinflammatory cytokines [IL-6, IL-1β, TNF-α, MIP-2] in the bronchoalveolar lavage fluid [BALF]), histologic lung injury (lung injury score), permeability of the alveolar capillary barrier (lung wet/dry weight ratio and BALF protein concentration) and mortality rates were attenuated in the PALM3 siRNA-treated mice. These results indicate that PALM3 contributes to the development of ALI in mice challenged with LPS. Inhibiting PALM3 through the intranasal application of specific siRNA protected against LPS-induced ALI.     

Urinary desmosine as a biomarker in acute lung injury

Acute lung injury (ALI) is a complex disorder associated with an acute inflammatory response thought to contribute to tissue injury. Desmosine, a cross-linking amino acid present in elastin, is released during matrix degradation and cleared by the kidney. Results from animal models and human disease studies have suggested that ALI is associated with the release of desmosine, resulting in increased urinary desmosine. A radioimmunoassay was used to monitor urinary desmosine levels over 10 days in ten patients with ALI. The concentration of desmosine was measured with and without acid hydrolysis. Baseline urinary desmosine was increased in two of ten patients. The concentration of desmosine at baseline did not appear to be related to age, gender, neutrophil elastase (NE)/α₁-antiprotease complex concentration or P_aO₂/F_iO₂ ratio. No meaningful changes in desmosine levels were noted after removal from mechanical ventilation. Baseline desmosine concentrations did not appear to correlate with the risk of death. The limited sensitivity, predictive correlations and dynamic modulation would suggest that urine desmosine has a limited role as a biomarker for ALI. Hydrolysis of urine samples appears necessary for optimal measurement of urine desmosine.