吡咯烷二硫代氨基甲酸对大鼠急性肺损伤TNF-α的影响

目的 探讨核因子-кB(NF-кB)抑制剂(吡咯烷二硫代氨基甲酸,PDTC)在脂多糖(LPS)所致大鼠急性肺损伤(ALI)中对肿瘤坏死因子-α (TNF-α)的影响.方法 雄性大鼠66只,随机分3组.对照组(N组)6只,腹腔注射生理盐水3 mL/kg; ALI组(L组)30只,腹腔注射LPS3 mg/kg; PDTC干预组(P+L组)30只,先腹腔注射PDTC120 mg/kg,半小时后再腹腔注射LPS3 mg/kg.后两组分别于腹腔注射LPS后1、2、4、8、12 h作为观测时间点.观察肺组织匀浆及血浆中TNF-α的变化.结果 L组各时相肺组织匀浆及血浆中TNF-α较N组升高(P＜0.01),且以2h组升高最为显著,但P+L组TNF-α与L组比较减少(P＜0.05).结论 LPS引起肺组织和血中TNF-α大量释放,NF-кB参与炎症的调控,在ALI中起重要作用.PDTC通过调控炎性介质的表达和释放,可有效地减轻LPS所致大鼠ALI.     

Effect of captopril on serum TNF-α level in acute lung injury rats induced by HCL

Objective:To observe the effect of captopril on the tumor necrosis factor-α(TNF- α) level and arterial blood gases in acute lung injury(All) induced by HCL in rats,and to analyze its protective mechanism.Methods:Fifty Wistar rats were selected and randomly divided into three groups,with 20 rats in Group Ⅰ and Ⅱ,respectively and 10 animals in Group Ⅲ.ALI model was constructed by intratracheal injection of diluted hydrochloric acid(pH=1.25.1.2 mL/kg).Group Ⅰrats received not any treatment after construction of AM model.Group Ⅱ rats were treated with captopril(5 mg/kg,i.p.) 5 min after induction of ALI.Group Ⅲ served as normal control without any treatment.Ninety minutes after construction of ALI model,all the rats were sacrificed.Blood was withdrawn for detection of TNF- α level and arterial blood gases index.And lung tissue slices of the three groups were prepared for observation of pathologic histology changes.Results:TNF- α level in serum of Group Ⅰ and Ⅱ rats was significantly higher than that in Group Ⅲ(P0.05),while TNF- α level in serum of Group Ⅱ was significantly lower in Group Ⅰ(P0.05).PaCO_2 level was significantly higher(P0.05),while PaO_2 was significantly lower(P0.05) in Group Ⅰ and Ⅱ rats than those in Group Ⅲ.PaCO_2 was significantly lower(P0.05) and PaO_2 was significantly higher(P0.05) in Group Ⅱ than those in Group Ⅰ.Histological observation showed diffuse congestion and severe edema of luug tissue,obvious thickening and structure damage of alveolar walls and a large amount of neutrophil infiltration in Group Ⅰ rats.Group Ⅱ rats showed mild edema of lung tissue;only a small portion of alveolar walls showed thickening and only a few of neutrophil infiltration could be observed.The degree of injury was remarkably slighter than that of Group Ⅰ rats.Group Ⅲ rats showed clear lung tissue structure and normal morphology:alveolar walls were uniform and the margin was smooth and few neutrophil could be observed.Conclusions:Captopril can significantly reduce serum TNF- α level,elevate PaO_2 and reduce PaCO_2 in rats with ALI.It has a protective effect on ALI rats.     

人脐带间充质干细胞对ALI治疗作用的研究

目的探讨输入人脐带间充质干细胞（human umbilical cord mesenchymal stem cells,HUMscs）对脂多糖（LPS）诱导的大鼠ALI的治疗作用。方法将30只Wistar大鼠随机分为3组：正常对照组、ALI模型组和HUMSCs移植组,每组10只。健康大鼠LPS气管滴入方法建立ALI模型,尾静脉输入HUMSCs。输入7h后,处死各组大鼠,取肺组织行病理学观察,测定肺湿／干重比（W／D）,ELISA法测定肺组织匀浆中肿瘤坏死因子α（TNF-α）和IL-1β的含量。结果肺组织病理学显示：ALI模型组大鼠肺组织呈现典型的炎症病理变化,包括肺泡充血、水肿、出血,肺泡腔和血管壁中性粒细胞浸润,肺泡壁增厚等肺损伤性病变;HUMSCs移植组大鼠肺组织损伤程度明显减轻。与正常对照组比较,ALI模型组W／D、TNF-α和IL-1β含量明显升高（P〈0．05）。与Au模型组比较,HUMSCs移植组W／D、TNF-α和IL-1β含量明显降低（P〈0．05）。结论输入HUMSCs能减轻ALI,并能降低肺W／D和肺组织TNF-α及IL-1β含量。     

DAPK1在急性肺损伤小鼠肺组织中的表达及其作用

目的探讨死亡相关蛋白激酶1(DAPK1)在急性肺损伤小鼠肺组织中的表达及在炎症失控中的作用。方法将30只雄性C57小鼠按随机数字表法分为5组:正常对照组、LPS诱导急性肺损伤3、6、12、24 h组。应用2 mg/kg DAPK1抑制剂TC-DAPK 6预处理小鼠后,再用10 mg/kg LPS诱导小鼠肺损伤。HE染色光镜观察小鼠肺组织病理改变;免疫组化检测肺组织中DAPK1的表达和分布;应用RT-PCR和Western blot检测肺组织DAPK1和NF-κB p65的表达;ELISA检测血清炎症因子TNF-α、IL-6水平变化;Kaplan-Meier生存分析对各组小鼠存活时间进行分析。结果 LPS致伤组肺组织可见明显病理损伤改变且随时间进展加重,而DAPK1蛋白在正常对照组小鼠肺组织仅少量表达,在急性肺损伤小鼠肺组织中表达随时间进展明显增加;与正常对照组相比,LPS组肺组织DAPK1 mRNA蛋白表达水平均明显增高(P0.05),血浆炎症因子TNF-α、IL-6均明显升高(P0.05)。抑制小鼠肺DAPK1表达可明显降低LPS诱导的肺组织中DAPK1和NF-κB p65蛋白水平、血清炎症因子TNF-α、IL-6的水平(P0.05)。此外,抑制DAPK1可延长LPS致急性肺损伤小鼠的生存期(P0.01)。结论 DAPK1通过调控NF-κB炎症通路参与了LPS诱导的小鼠急性肺损伤,其可作为潜在的治疗靶点。     

Effect of sevoflurane on tissue permeability of lung ischemia-reperfusion injury in rats

Objective:To investigate the effect of sevoflurane on tissue permeability of lung ischemiareperfusion injury(LIRI)in rats.Methods:A total of 45 wistar rats were randomly divided into3 groupsⅠ,Ⅱ,Ⅲ.Modified Eppinger method was adopted to establish the rat lung ischemiareperfusion injury model.GroupⅠserved as the control group,groupⅡas ischemia reperfusion group,groupⅢas sevoflurane ischemia-reperfusion group.Blood gas index,lung permeability index(LPI)change,lung tissue pathology change and lung water content were observed and compared between groups of rats at different time points.Results:During ischemia reperfusion,all rats kept balance of the MAP during different time points,SPO_2 of groupⅡandⅢdecreased significantly thanⅠgroup(P0.05);after reperfusion lung permeability index in GroupⅡandⅢwas higher than the control group significantly(P0.05),120 min after reperfusion LPI change and iujury of groupⅢwas significantly lower thanⅡgroup(P0.05);interstitial and alveolar cavity effusion in of groupⅢwere lower than that of groupⅡ.Conclusions:Sevoflurane pretreatment can reduce the lung tissue permeability,and LIRI plays a protective role in LIRI.     

Exogenous porcine surfactants increase the infiltration of leukocytes in the lung of rats

BackgroundSeveral studies have investigated the influence of exogenous surfactants on inflammatory response in the lung, however results reported about effects of surfactants on the lung infiltration of leukocytes are controversial. Our previous study noticed that treatment of porcine surfactant (PS) significantly increased the lung infiltration of leukocytes in rats with acute lung injury (ALI). The objective of this study was to verify the effect of exogenous PS on the lung infiltration of leukocytes in vivo and investigate the possible mechanisms involved in vitro.MethodsThe number of leukocytes in bronchoalveolar lavage fluid (BALF) of rats with or without lipopolysaccharide (LPS)-induced ALI was determined after treatment with different concentrations of PS, dexamethasone (Dex) or PS + Dex. The effect of PS and Curosurf, a commercially available porcine surfactant, on human peripheral neutrophil migration was determined by the Boyden Chamber Assay.ResultsInstillation of PS significantly increased the number of leukocytes in BALF of normal rats and rats with LPS-induced ALI. Most of the increased leukocytes were neutrophils. Dex significantly decreased the number of leukocytes and TNF-α concentration in BALF caused by LPS, but did not significantly reduce the number of leukocytes increased by PS. In vitro experiments further demonstrated that both PS and Curosurf had direct chemotactic effects on neutrophils.ConclusionsThese results suggest that PS contain chemoattractant(s) which induce the infiltration of leukocytes, especially neutrophils, into lung.     

气管内滴入核仁素shRNA表达载体对脂多糖肺损伤大鼠肺泡巨噬细胞活化的影响

目的建立气管内滴人核仁素shRNA表达载体的实验模型,观察其对脂多糖（LPS）诱导急性肺损伤（ALI）大鼠肺泡巨噬细胞（AM）活化的影响,探讨AM膜核仁素在LPS致大鼠ALI中的作用及机制。方法选择健康雄性SD大鼠40只,随机分为A组（空白对照组）,B组（ALI组）,C组（转染重组质粒pBS-U6-C23shRNA并ALI组）,D组（转染对照质粒pBS-U6-ControlshRNA并ALI组）。大鼠经尾静脉注射LPS以建立Au模型,气管内滴人法转染重组质粒pBS-U6-C23shRNA表达载体,免疫印迹（Westernblot）检测AM膜核仁素蛋白表达;核酸电泳迁移率实验（EMSA）检测AM中核因子（NF-κB）活性;酶联免疫吸附法（ELISA）检测肺泡灌洗液（BALF）中肿瘤坏死因子-α（TNF-α）和白介素-6（IL-6）表达。结果转染pBS-U6-C23shRNA表达载体后可以显著下调AM膜核仁素蛋白表达;减轻肺病理损伤;降低NF-κB的活性以及肺泡灌洗液中TNF-α和IL-6的含量。与A组相比,B组及D组的大鼠AM膜核仁素蛋白表达显著增加,组间有统计学差异（P〈0．01）;C组大鼠AM膜核仁素蛋白表达明显下调,与B组相比,组间差异有统计学意义（P〈0．01）;而D组与B组中的大鼠AM膜核仁素蛋白表达组间没有统计学差异（P〉0．05）。经LPS处理后,各组大鼠AM中NF-κB活性显著增加,与A组相比,组间差异非常明显（P〈0．01）。c组大鼠AM中NF-κB活性明显降低,与B组比较,差异显著（P〈0．05）;而D组与B组比较,组间无明显差异（P〉0．05）。经LPS诱导肺损伤后,显著增加了大鼠BALF中促炎因子TNF-α和IL-6的表达,各组和A组相比差异显著（P〈0．01）;C组大鼠BALF中TNF-α和IL-6的含量显著降低,与B组相比差异显著（P〈0．01）;而D组与B组相比,组间差异无统计学意义（P〉0．05）。结论气管内滴入法可以安全有效转染核仁素shRNA表达载体,抑制AM活化,减轻肺损伤,AM膜核仁素参与了LPs致大鼠ALI的发病过程。     

Evaluation of lung injury and respiratory mechanics in a rat model of acute pancreatitis complicated with endotoxin

BackgroundAcute lung injury (ALI) is a common complication of acute pancreatitis (AP) and contributes to the majority of AP-associated deaths, particularly in the setting of secondary infection. This ‘two-hit’ model mimics clinical cases where the presentation of AP is associated with mild lung injury that, following a secondary direct lung infection, can result in respiratory dysfunction and death. We therefore aimed to characterize lung injury in a clinically-relevant ‘two-hit’ rat model of caerulein-induced AP combined with intratracheal endotoxin.MethodsRats received 7 hourly intraperitoneal injections of caerulein (50 μg/kg). Twenty four hours following the first caerulein injection, rats were anaesthetised and LPS (15 mg/kg) was instilled intratracheally. Following LPS instillation, rats were ventilated for a total of 2 h.ResultsIn the present study, AP results in mild pulmonary injury indicated by increased lung myeloperoxidase (MPO) activity and edema, but with no alteration of respiratory function, while intratracheal instillation of LPS results in more substantial pulmonary injury. The induction of AP challenged with secondary intratracheal LPS results in an exacerbation of lung damage indicated by further increased lung edema, plasma and bronchoalveolar (BAL) CINC-1 concentration, lung damage histology score, and lung tissue resistance and elastance, compared with LPS alone.ConclusionsIn conclusion, the addition of instilled LPS acted as a “second-hit” and exacerbated caerulein-induced AP, compared with the induction of AP alone or the instillation of LPS alone. Given its clinical relevance, this model could prove useful for examination of therapeutic interventions for ALI following secondary infection.     

Contributions of high mobility group box protein in experimental and clinical acute lung injury

This study was performed to examine the putative role of high mobility group box (HMGB) protein in the pathogenesis of acute lung injury (ALI). Observations were made (1) in 21 patients who were septic with ALI and 15 patients with normal lung function and (2) in a mouse model 24 hours after intratracheal instillation of lipopolysaccharide (LPS). The concentrations of HMGB1 were increased in plasma and lung epithelial lining fluid of patients with ALI and mice instilled with LPS. LPS-induced ALI was mitigated by anti-HMGB1 antibody. Although this protein was not detected in the plasma of control humans or mice, the concentrations of HMGB1 in lung epithelial lining fluid or in bronchoalveolar lavage fluid were unexpectedly high. The nuclear expression of HMGB1 was apparent in epithelial cells surrounding terminal bronchioles in normal mice, whereas its nuclear and cytoplasmic expression was observed in alveolar macrophages in LPS-instilled mice. Lung instillation of HMGB2 did not cause as much inflammation as HMGB1. Extracellular HMGB1 may play a key role in the pathogenesis of clinical and experimental ALI. However, its expression in normal airways is noteworthy and suggests that it also plays a physiologic role in the lung.     

Immunohistochemical study of tumor necrosis factor-alpha in acute liver injury induced by Propionibacterium acnes and lipopolysaccharide in rats

The effect of intravenous injection of Propionibacterium acnes (P. acnes) and lipopolysaccharide (LPS) on the distribution of tumor necrosis factor-α (TNF-α) in different organs have not previously been investigated. Immunohistochemistry and histological examination were employed in evaluating the distribution of TNF-α in the liver, spleen, lungs and bone marrow in rats injected intravenously with P. acnes followed by LPS 7 days later. Granulomas containing ED1-positive macrophages were observed in the liver 7 days after P. acnes injection. Subsequent LPS injection resulted in proliferation of ED1-positive macrophages in the sinusoids and coagulation necrosis of hepatocytes after 6 h. TNF-α was detected in ED2-positive macrophages (Kupffer cells) 1 day after P. acnes injection and in macrophages constituting the granulomas 7 days later, but prior to LPS injection. TNF-α was also detected in ED1-positive macrophages in the spleen, predominantly in the marginal zone. When granulomas were formed 7 days after P. acnes injection, TNF-α was observed in macrophages of the granulomas. TNF-α was also detected in macrophages of the granulomas found in the lung 1 day after P. acnes injection. No macrophages expressing TNF-α were found in the granulomas of bone marrow. The highest expression was in the liver at any time interval and in macrophages constituting granulomas. Our results suggest that the high expression of TNF-α in the liver results in selective hepatic necrosis. The expression of TNF-α in macrophages of the liver after P. acnes injection and the subsequent development of hepatic necrosis after LPS injection suggest that P. acnes acts as an inducer of TNF-α production in macrophages while LPS acts as a trigger for the release of TNF-α from macrophages.     

Tumor Necrosis Factor-α Plays an Initiating Role in Extracorporeal Circulation-induced Acute Lung Injury

Background

Despite advances in critical care, the mortality rate for patients with acute lung injury (ALI) remains high. The aim of this study was to test the hypothesis that tumor necrosis factor-α (TNF-α) plays an initiating role in the onset of extracorporeal circulation (ECC)-induced ALI.

Methods

Eight New Zealand rabbits subjected to 1 h of ECC and 40 min of observation after termination of ECC were used for monitoring pulmonary nociceptor activity. Fifty Sprague-Dawley (SD) rats that received 2 h of ECC and 4 h of rest were used to measure the pulmonary function and inflammatory cytokines release, including total cells, neutrophils, and TNF-α in bronchoalveolar lavage (BAL) and white blood cell (WBC) and neutrophils in blood. An additional 40 SD rats were randomized to pretreatment with inhalation of phosphate buffer solution (control group), IgG (IgG inh group), or TNF-α antibody (anti-TNF-α inh group) and venous injection of TNF-α antibody (anti-TNF-α iv group). After 2 h of ECC and 4 h of rest, the arterial blood and BAL fluid were collected for measurement of arterial oxygen pressure (PaO₂) and inflammatory cytokines release. The left-lower-lung tissues of animals were stained with hematoxylin & eosin (H&E).

Results

The results demonstrated that the activities of airway nociceptor and TNF-α release were similarly upregulated at the early stage and in a time-related manner in ECC-induced ALI. Pretreatment with TNF-α antibody inhalation, but not venous injection, improved pulmonary function, inhibited pulmonary inflammation, and attenuated pulmonary histopathological changes after ECC.

Conclusion

We concluded that TNF-α played an important role in the pathogenesis of ALI and acted as an initiating cytokine at the early stage of ECC-induced ALI.     

lncRNA PACER促进脓毒症急性肺损伤炎症反应的实验研究

目的探讨lncRNA PACER对小鼠脓毒症急性肺损伤炎症反应的促进作用。 方法分离和培养急性肺损伤和健康非吸烟者的肺泡巨噬细胞,以及获取细菌脂多糖(LPS)诱导的急性肺损伤(ALI)小鼠肺组织,采用qRT-PCR方法检测lncRNA PACER的表达;过表达或敲低PACER后,ELISA方法检测THP-1和RAW264.7细胞炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的表达;对LPS所致ALI小鼠,尾静脉注射PACER siRNA慢病毒后,ELISA检测小鼠血清炎性因子TNF-α、IL-6的表达,HE染色观察肺组织病理学变化。 结果ALI患者肺泡巨噬细胞和ALI小鼠肺组织中lncRNA PACER表达均显著升高(P<0.01);细胞过表达PACER后,细胞炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的表达升高,而敲低PACER后,炎性因子TNF-α、IL-6的表达则降低(P<0.01);ALI小鼠敲低PACER后,小鼠肺组织和血清中炎性因子TNF-α、IL-6的表达均显著降低(P<0.01),肺组织损伤程度明显减弱。 结论lncRNA PACER可显著促进脓毒症急性肺损伤炎症反应,为明确lncRNA PACER作为ALI防治的靶标提供了依据。     

Expression change of TNF–α in myocardium and hepatic tissue of rats with compound stress of hyperthermia and lipopolysaccharide

ObjectiveTo investigate the expression characteristics of TNF-α in myocardium and hepatic tissue of rats with compound stress of hyperthermia and lipopolysaccharide (LPS).MethodsMale SPF Wistar rats were randomly divided into room temperature+physiological saline group (Group C), hyperthermia+physiological saline group (Group H), room temperature+LPS group (Group L) and hyperthermia+LPS group (Group HL). The rats were put in simulated climate cabin. Group HL and Group H were exposed in the environment at a dry bulb temperature (TDB) of (35.0±0.5) °C, while Group L and Group C were exposed in the environment at a TDB of (26.0±0.5) °C. The rats in Group HL and Group L were given tail intravenous injection of LPS 10 mg/kg, while the rats in Group H and Group C were given tail intravenous injection of 9 g/L NaCl 10 mL/kg. After the stress, immunohistochemical SABC staining method was used to detect the expression characteristics of TNF-α in myocardium and hepatic tissue of rats, and those rats were given routine pathological examinations.ResultsThe expression of TNF-α in myocardium and hepatic tissue in Group HL was enhanced remarkably, and the tissue damages of Group HL were severest.ConclusionsThe eardiotoxieity and hepatotoxicity caused by compound stress of hyperthermia and LPS is closely related to the expression of TNF-α.     

内毒素致大鼠急性肺损伤模型的建立

目的建立经尾静脉注射脂多糖致大鼠急性肺损伤动物模型。方法大鼠尾静脉注射5 mg/kg脂多糖,造成急性肺损伤,于注射脂多糖2 h后,分别测定动脉血气分析、血清蛋白含量、肺泡灌洗液中蛋白、细胞间粘附分子-1的含量、肿瘤坏死因子-α、白介素-8,计算肺湿/干重比、肺通透指数,观察肺组织病理学改变。实验设置生理盐水对照组。结果肺组织病理切片提示脂多糖组肺间质大量炎性细胞浸润,出血、水肿,而生理盐水组肺组织损伤轻微。对照组肺湿/干重比、肺泡通透指数、肺泡灌洗液中中性粒细胞比均显著低于脂多糖组(P<0.05),而动脉血氧分压则高于脂多糖组(P<0.05)。脂多糖组肺泡灌洗液中肿瘤坏死因子-α、细胞间粘附分子-1、白介素-8浓度高于对照组(P<0.05)。结论此模型基本符合急性肺损伤诊断标准及动物模型的考察指标符合临床,说明本模型是成功可靠的。     

乙酰化白藜芦醇通过调节缝隙连接蛋白43表达减轻海水淹溺型急性肺损伤

目的观察海水淹溺肺损伤大鼠肺组织中缝隙连接蛋白43（Cx43）的表达变化及乙酰化白藜芦醇对其的干预作用。方法32只大鼠完全随机分为空白对照组、模型组、乙酰化白藜芦醇预处理组和乙酰化白藜芦醇组,每组8只。采用气管内滴注海水（4ml／kg）的方法制作海水淹溺型急性肺损伤大鼠模型,用ELISA法检测肺组织中TNF-仅和IL-10的含量,实时定量PCR及免疫组化的方法检测肺组织中Cx43的表达变化。结果吸人海水4h后,大鼠肺部损伤明显,TNF—α和IL-10的含量增高,Cx43在基因水平表达量增高,而蛋白水平表达降低;乙酰化白藜芦醇预处理明显减轻海水吸入导致的急性肺损伤,减少炎症因子分泌并且在基因水平和蛋白水平增加Cx43的表达。结论Cx43参与海水淹溺型急性肺损伤的发生与发展,乙酰化白藜芦醇能够通过增加Cx43的表达减轻肺损伤。     

氨溴索在保护氟康唑所致大鼠肺组织损伤中的作用及机制研究

目的 研究氨溴索对气管内注射氟康唑所致大鼠肺组织损伤的保护作用及肿瘤坏死因子α(TNF-α)在肺组织中的表达变化及意义.方法 将90只雄性SD大鼠随机平均分成3组,分别为正常对照组(N组)、氟康唑组(F组)、氟康唑+盐酸氨溴索干预组(FA组).除N组外均行气管穿刺注入氟康唑,FA组气管内给药前6h腹腔注射氨溴索,于6个时间点(2h、1d、2d、3d、5d、7 d)分别处死大鼠5只,取右肺副叶行HE染色观察肺组织病理变化和免疫组化检测肺组织TNF-α蛋白的表达变化.结果 病理结果提示F组和FA组肺泡内均有中性粒细胞浸润,F组气管、血管周围及肺泡内炎症细胞浸润最明显.免疫组化提示F组2h时染色最浅,TNF-α蛋白的表达从2h开始至第3天逐渐增强,第3天时着色最深;F组各时间点TNF-α蛋白染色程度评分均高于FA组和N组,差异有统计学意义(P值均＜0.01).结论 气管内局部使用氟康唑可对大鼠肺组织造成损伤,并使肺组织TNF-α蛋白的表达升高,氨溴索对该损伤具有保护作用,可能与下调TNF-α蛋白的表达相关.     

Resveratrol inhibits cell apoptosis by suppressing long noncoding RNA (lncRNA) XLOC_014869 during lipopolysaccharide-induced acute lung injury in rats

BackgroundAcute lung injury (ALI) is a common clinical complication with a high mortality rate. Resveratrol (Res) has been shown to protect against ALI, but the role of long noncoding RNAs (lncRNAs) in this process is still unclear.MethodsMale rats (n=20) aged 7–8 weeks were randomly divided into four groups: control, lipopolysaccharide (LPS), LPS + Res, and LPS + dexamethasone (Dexa). Intragastric administration of Res (0.5 mg/kg) or Dexa (1.5 mg/kg) was performed 1 h before intraperitoneal injection of LPS (5 mg/kg). Lung tissue, serum, and bronchoalveolar lavage fluid were sampled 6 h after LPS treatment for inflammatory factor detection, pathological detection, lncRNA sequencing and bioinformatical analysis, and TdT-mediated dUTP Nick-End Labeling. Quantitative real time polymerase chain reaction and western blotting were used to verify the sequencing results. LPS, Res, and RNA interference were used in rat alveolar epithelial cells experiments to confirm the protective of Res/lncRNA against ALI.ResultsRes pretreatment inhibited lung injury and the increase of inflammatory cytokines induced by LPS. The differentially expressed lncRNAs and mRNAs (P<0.05 and |fold change| >2) were mainly involved in the signaling pathway of immunity, infection, signaling molecules and interactions. Among the lncRNAs and mRNAs, 26 mRNAs and 23 lncRNAs had high levels in lungs treated with LPS but decreased with Res, and 17 mRNAs and 27 lncRNAs were at lower levels in lungs treated with LPS but increased with Res. lncRNA and adjacent mRNA analysis showed that lncRNAs XLOC_014869 and the adjacent gene Fos, and the possible downstream genes Jun and Faslg were increased by LPS, but these changes were attenuated by Res. Pretreatment with Res reduced LPS-induced lung tissue apoptosis. Similarly, Res treatment and knockdown of lncRNA XLOC_014869 reduced LPS-induced apoptosis and the levels of Fos, c-Jun, and Fas-L.ConclusionsRes can inhibit the increase of lncRNAs XLOC_014869 caused by LPS stimulation and inhibit lung cell apoptosis. These effects may be due to lncRNA XLOC_014869 mediation of the pro-apoptotic factors (Fos, c-Jun, and Fas-L).     

Small volume resuscitation with 7.5% hypertonic saline, hydroxyethyl starch 130/0.4 solution and hypertonic sodium chloride hydroxyethyl starch 40 injection reduced lung injury in endotoxin shock rats: comparison with saline

BackgroundThe aim of this study was to investigate the effects of small volume resuscitation with 7.5% hypertonic sodium chloride (HSS), hydroxyethyl starch 130/0.4 solution (HES), and hypertonic sodium chloride hydroxyethyl starch 40 injection (HSH) on endotoxin shock rat lung.MethodsThirty Sprague–Dawley (SD) rats were divided randomly into 5 groups ,Group C (negative control group), Group E (lipopolysaccharide, LPS +4 ml/kg saline), Group HSS (LPS +4 ml/kg HSS), Group HES (LPS +4 ml/kg HES) and Group HSH (LPS +4 ml/kg HSH). Endotoxin shock model of rat was produced by injection with LPS. Then small volume resuscitation with different fluids was implemented in each group, respectively.ResultsCompared to Group C(negative control group）, lung injury in the other four groups was increased. Compared to Group E(LPS +4 ml/kg normal saline), lung injury of Group HSS(LPS +4 ml/kg HSS), HES(LPS +4 ml/kg HES), and HSH (LPS +4 ml/kg HSH)was lessened. Compared to Group C, oxygenation index in Groups E, HSS, HES, and HSH were decreased (P < 0.01). Compared to Group E, oxygenation indexes in Groups HSS, HES, and HSH were significantly increased (P < 0.01). Data of tumor necrosis factor (TNF)-α of lung tissue had similar results. However, protein concentration of bronchoalveolar lavage fluid and hydrogen sulfide (H₂S) concentration indicated contrary results.ConclusionSmall volume resuscitation with 7.5% hypertonic sodium chloride, hydroxyethyl starch 130/0.4 solution, and hypertonic sodium chloride hydroxyethyl starch 40 injection could lessen lung injury caused by lipopolysaccharide. And this effect had relation to change of TNF-α and H₂S.     

红景天苷对脂多糖所致急性肺损伤治疗作用的研究

目的:观察红景天苷（SDS）对脂多糖（LPS）引起的大鼠急性肺损伤（ALI）的治疗作用,并初步探讨其作用的机制。方法: 将36只SD大鼠随机分为生理盐水（NS）对照组、LPS组和LPS+SDS组(每组n=12),通过向气管内滴注LPS建立大鼠ALP模型。采用微量注射泵向大鼠右颈外静脉注射液体并留置的给药方法,NS组和LPS组注射NS并留置4 h;LPS+SDS组先于气管内滴注LPS,0.5 h后再注射SDS并留置4 h。4.5 h后处死实验动物,取肺组织观察其形态学改变,测定肺湿/干质量的比值（W/D）、肺泡灌洗液（BALF）中蛋白的含量及肺组织匀浆中TNF-α、IL-6、IL-10、乳酸脱氢酶(LDH)和髓过氧化物酶(MPO)的含量。结果: 形态学观察表明,LPS组肺组织水肿,有点、片状出血及大量的炎性细胞浸润,肺泡间隔显著增厚,肺泡腔变窄、结构消失。LPS+SDS组的肺组织损伤明显减轻,肺组织结构趋于正常,肺泡腔及支气管腔炎细胞及渗出物与LPS组相比明显减少。LPS组肺W/D与NS组相比明显增加（P<0.05）,BALF中蛋白的含量显著增加（P<0.05）,肺组织匀浆中TNF-α、IL-6、IL-10、LDH和MPO的含量显著增加（P<0.05）;而给予SDS治疗后,肺W/D、BALF中蛋白含量、肺组织匀浆中TNF-α、IL-6、LDH和MPO的含量与LPS组相比均明显减少（P<0.05）,IL-10含量显著增加（P<0.05）。结论: SDS对LPS所致ALI具有治疗作用,这种治疗作用可能与其通过抑制肺组织中炎症介质的作用有关。     

Knockdown of myeloid differentiation protein-2 reduces acute lung injury following orthotopic autologous liver transplantation in a rat model

BackgroundAcute lung injury (ALI) is a serious complication that commonly occurs during orthotopic liver transplantation (OLT). Toll-like receptor 2/4 (TLR2/4) are the main membrane receptors that respond to inflammatory stimuli and mediate NF-kappa B (NF-κB) signal pathway. We previously showed that TLR2/4 expression on monocytes and serum cytokine levels were increased in patients with ALI induced by OLT. Myeloid differentiation protein-2 (MD-2) expresses the functional domains that combines TLRs and play a key regulatory role in TLRs activation. Therefore, we hypothesized that blocking MD-2 would inhibit the TLR2/4-mediated inflammatory response and lessen ALI induced by liver transplantation.MethodThirty-two Sprague Dawley (SD) rats were randomly divided into four groups. One group received a sham operation (Group S), and the other three groups underwent orthotopic autologous liver transplantation (OALT) 48 h after intratracheal administration of saline (Model group; Group M), non-targeting siRNA (negative siRNA control group; Group NC) or siRNA against MD-2 (intervention group; Group RNAi). Lung pathology, lung water content, PaO₂, and expression levels of MD-2, TLR2/4, NF-κB, TNF-α, IL-1β and IL-6 were assessed 8 h after OALT.ResultsIn Groups M and NC, OALT produced marked lung pathology with decreased PaO₂ levels and increased MD-2, TLR2/4 gene and protein expression levels. Furthermore, the nuclear translocation of the NF-κB P65 subunit, was increased, as were lung concentrations of TNF-α, IL-1β and IL-6. The pathology of ALI and the severity of the above biochemical changes induced by OALT were significantly reduced in the group treated with MD-2 siRNA.ConclusionMD-2 gene knock-down attenuated the increase in TLR2/4 activation and reduced ALI after OALT.