HO-1在CCK-8减轻脂多糖所致的急性肺损伤中的作用

目的： 探讨血红素氧合酶（HO）-1在八肽胆囊收缩素（CCK-8）减轻脂多糖（LPS）所致急性肺损伤（ALI）中的作用。方法: 将大鼠随机分为5组：正常对照组、LPS组、CCK-8+LPS组、LPS+Hm（氯血红素，CO供体）组、LPS+ZnPP（锌原卟啉，HO-1特异性阻断剂）组。各组给药后2 h、6 h、12 h行支气管肺泡灌洗、检测支气管肺泡灌洗液（BALF）中中性粒细胞（PMN）数目；进行肺组织的形态学观察；测定肺组织中丙二醛（MDA）含量和HO-1蛋白活性；应用RT-PCR和Western blotting技术检测给药后6h肺组织中HO-1 mRNA和蛋白的表达情况。结果: LPS组肺组织出现损伤性变化，同时BALF中PMN数目、肺组织中MDA含量、HO-1蛋白活性、HO-1 mRNA和蛋白的表达均高于相应对照组（均P<0.05）；CCK-8+LPS和LPS+Hm组肺组织损伤程度、BALF中PMN数目和肺组织中MDA含量低于相应LPS组，而肺组织中HO-1蛋白活性、HO-1 mRNA和蛋白的表达均高于相应LPS组（均P<0.05）；LPS+ZnPP组肺组织损伤程度、BALF中PMN数目和肺组织中MDA含量分别高于相应LPS组，而肺组织中HO-1蛋白活性、HO-1 mRNA和蛋白的表达分别低于相应LPS组（均P<0.05）。结论: CCK-8可部分通过HO-1介导的抗氧化、抑制PMN聚集等效应来发挥减轻LPS所致的肺损伤作用。     

小窝蛋白1脚手架区多肽减轻LPS诱导的小鼠急性肺损伤

目的:探究人工合成的小窝蛋白1(caveolin-1,Cav-1)脚手架区多肽cavtratin对血红素加氧酶1(heme oxygenase-1,HO-1)活性及脂多糖(lipopolysaccharide,LPS)诱导的小鼠急性肺损伤的作用。方法:成年雄性BALB/c小鼠随机分为6组,每组8~10只,实验分为对照(control)组、触足肽内化序列(Antennapedia internalization sequence,AP)组、LPS组、LPS+血晶素(hemin)组、LPS+hemin+cavtratin组和LPS+hemin+cavtratin+锌原卟啉(zinc protoporphyrin IX,Zn PP)组。小鼠气管滴注LPS 24 h后,苏木素-伊红染色观察肺组织病理形态变化;检测肺组织湿/干重比、肺泡灌洗液中细胞数和血清中乳酸脱氢酶活性。免疫荧光观察HO-1和Cav-1的结合情况并检测HO-1活性;实时荧光定量PCR检测炎症因子(IL-1β、IL-6、TNF-α、MCP-1和i NOS)的mRNA水平。结果:组织免疫荧光以及HO-1活性检测发现,与LPS组比较,LPS+hemin+cavtratin组HO-1与细胞膜Cav-1的结合减少,HO-1的活性增高(P0.05);与LPS组比较,LPS+hemin+cavtratin组肺组织受损程度明显减轻,肺湿/干重比、肺泡灌洗液细胞数和血清中乳酸脱氢酶活性显著降低(P0.05);与LPS组比较,LPS+hemin+cavtratin组炎症因子的mRNA表达降低(P0.05);HO-1活性抑制剂Zn PP可以消除cavtratin的保护作用。结论:Cavtratin可以通过减少HO-1与细胞膜Cav-1的结合使得HO-1活性增加,进而减轻LPS诱导的小鼠急性肺损伤。     

亚精胺减轻脂多糖诱导的小鼠急性肺损伤

目的　观察亚精胺（spermidine）对脂多糖（LPS）诱导的急性肺损伤（ALI）的影响。 方法　采用5 mg/kg的LPS经气管滴注,建立ALI小鼠模型。用小动物呼吸机检测亚精胺对ALI小鼠呼吸功能的影响;观察亚精胺对ALI小鼠肺组织形态变化的影响;检测ALI小鼠支气管肺泡灌洗液（BALF）中总蛋白、总细胞数及中性粒细胞数目,并检测髓过氧化物酶（MPO）的水平;qPCR检测ALI小鼠肺组织中TREM-1 mRNA的表达;ELISA检测ALI小鼠BALF中sTREM-1的蛋白水平。 结果　亚精胺可改善ALI小鼠的呼吸功能;减轻LPS诱导的肺部病理损伤;减少蛋白渗出和中性粒细胞浸润;降低ALI小鼠肺内炎症放大受体TREM-1的表达。 结论　亚精胺能减少炎症细胞浸润,抑制炎症因子表达,从而减轻LPS诱导的ALI,其机制可能与亚精胺可抑制ALI小鼠肺组织TREM-1表达有关。     

亚精胺减轻脂多糖诱导的小鼠急性肺损伤

目的　观察亚精胺（spermidine）对脂多糖（LPS）诱导的急性肺损伤（ALI）的影响。 方法　采用5 mg/kg的LPS经气管滴注,建立ALI小鼠模型。用小动物呼吸机检测亚精胺对ALI小鼠呼吸功能的影响;观察亚精胺对ALI小鼠肺组织形态变化的影响;检测ALI小鼠支气管肺泡灌洗液（BALF）中总蛋白、总细胞数及中性粒细胞数目,并检测髓过氧化物酶（MPO）的水平;qPCR检测ALI小鼠肺组织中TREM-1 mRNA的表达;ELISA检测ALI小鼠BALF中sTREM-1的蛋白水平。 结果　亚精胺可改善ALI小鼠的呼吸功能;减轻LPS诱导的肺部病理损伤;减少蛋白渗出和中性粒细胞浸润;降低ALI小鼠肺内炎症放大受体TREM-1的表达。 结论　亚精胺能减少炎症细胞浸润,抑制炎症因子表达,从而减轻LPS诱导的ALI,其机制可能与亚精胺可抑制ALI小鼠肺组织TREM-1表达有关。     

白藜芦醇通过减弱中性粒细胞迁移和浸润改善LPS诱导的急性肺损伤

目的：探究白藜芦醇（RSV）通过减弱中性粒细胞迁移和浸润改善脂多糖（LPS）诱导的急性肺损伤（ALI）。方法 ：将BALB/c小鼠随机分为对照组、RSV组、模型组及模型+RSV组（LPS+RSV）,模型组和LPS+RSV组采用LPS诱导小鼠ALI模型,RSV组和LPS+RSV组以40 mg/kg RSV灌胃,连续灌胃7 d。末次给药后,收集小鼠支气管肺泡灌洗液（BALF）,检测BALF中白细胞、中性粒细胞和蛋白含量,评估肺组织髓过氧化物（MPO）活性,H-E染色观察肺组织病理损伤,检测肺组织和BALF中炎症细胞因子、趋化因子的表达。体外培养中性粒细胞,分别以1μg/mL LPS、1μg/mL RSV处理细胞,流式细胞术检测中性粒细胞凋亡、趋化因子受体（CXCR2、Mac-1）表达,活性氧荧光探针（DCFH-DA）检测细胞内活性氧（ROS）水平,评估中性粒细胞趋化性和迁移能力,免疫印迹检测核因子NF-κB p65磷酸化水平。结果：与模型组比较,LPS+RSV组BALF中细胞总数、中性粒细胞数、蛋白量、TNF-α、IL-6及CXCL2表达降低,肺组织MPO活性、TNF-α、IL-1β、IL...     

腺苷A2A受体激动剂CGS21680对脂多糖诱导的小鼠急性肺损伤的保护作用

 目的：探讨腺苷A2A受体激动剂对脂多糖(LPS)诱导的急性肺损伤（ALI）小鼠的作用。方法：采用LPS（10 mg/kg）气管内注射6 h后的ALI小鼠模型。实验动物随机分为生理盐水对照组、ALI组、CGS21680治疗组和CGS21680对照组。测定各组6 h后肺湿重/干重比值（W/D）。比色法测定各组肺组织中髓过氧化物酶(MPO)活性。Western blotting分析各组肺组织中细胞间黏附分子1（ICAM-1）和血管细胞黏附分子1（VCAM-1）的表达。酶联免疫吸附法（ELISA）检测各组小鼠血清中单核细胞趋化蛋白1（MCP-1）的浓度。观察各组肺组织病理改变。结果：ALI组肺W/D、MPO活性、ICAM-1及VCAM-1表达和MCP-1浓度均较对照组显著升高。CGS21680治疗组前述各项指标较ALI组有显著下降。CGS21680组较对照组各项指标无显著差异。肺组织病理显示,ALI组可见肺间质明显充血水肿,大量炎症细胞浸润,部分肺泡腔内可见红细胞。CGS21680治疗组可显著改善肺组织的病理变化。结论：腺苷A2A受体激动剂CGS21680可明显减轻LPS诱导的ALI小鼠肺组织的炎症反应及肺组织水肿,提示腺苷A2A受体激动剂在急性肺损伤中具有保护作用。     

硫化氢在脂多糖所致大鼠急性肺损伤中的作用

目的探讨硫化氢(H2S)在脂多糖(LPS)所致大鼠急性肺损伤(ALI)中的作用及可能的机制。方法将64只SD大鼠随机分为对照组、LPS组(经气管内滴注LPS复制ALI)、NaHS LPS组和炔丙基甘氨酸(PPG) LPS组。给药后4 h或8 h处死,测定肺系数;光镜观察肺组织形态学改变;化学法检测血浆H2S和CO含量、肺组织丙二醛(MDA)含量、胱硫醚-γ-裂解酶(CSE)和血红素加氧酶(HO)活性;用免疫组织化学法检测肺组织HO-1蛋白表达及吸光度值。结果气管内滴注LPS可引起肺组织明显的形态学改变;肺系数和肺组织MDA含量增加;血浆H2S含量和肺组织CSE活性下降;肺组织HO活性和HO-1蛋白表达增强;血浆CO含量增加。预先给予NaHS可显著减轻LPS所致上述指标的改变。而预先给予PPG可加重LPS所致肺损伤,但对CO含量、HO活性和HO-1蛋白表达无明显影响。结论H2S/CSE体系的下调在LPS所致ALI的发病学中有一定作用,而外源性给予一定量的NaHS对肺组织具有保护作用,该作用可能与其抗氧化效应和上调CO/HO-1体系有一定关系。     

内源性H2S在CCK-8减轻脂多糖所致急性肺损伤中的作用

目的：探讨内源性硫化氢(H₂S)在八肽胆囊收缩素(CCK-8)减轻脂多糖（LPS）所致急性肺损伤(ALI)中的作用。方法: 将84只SD大鼠随机分为正常对照组、LPS组（经气管内滴注LPS复制ALI）、NaHS(H₂S供体)+LPS组、炔丙基甘氨酸［胱硫醚-γ-裂解酶（CSE）抑制剂，PPG］+LPS组、CCK-8+LPS组、PPG+CCK-8+LPS组和CCK- 8组。给药后分别于4 h和8 h处死动物，测定肺湿/干比值；光镜观察肺组织形态学改变；化学法检测血浆H₂S含量，肺组织MDA含量、MPO活性和CSE活性；放免法检测肺组织P-selectin含量；RT-PCR检测肺组织CSE mRNA的表达；并行支气管肺泡灌洗，检测支气管肺泡灌洗液(BALF)中蛋白含量。结果: 气管内滴注LPS可引起肺组织明显的形态学改变；肺湿/干比值、BALF中蛋白含量及肺组织MDA、MPO活性和P-selectin水平增高；血浆H2S含量、肺组织CSE活性及CSE mRNA表达下降。预先给予NaHS或CCK-8可显著减轻LPS所致的上述改变，且血浆H₂S含量、肺组织CSE活性及CSE mRNA表达高于相应的LPS组；预先给予PPG可加重LPS所致的肺损伤，而血浆H₂S含量、肺组织CSE活性及CSE mRNA表达分别低于相应的LPS组和CCK-8+LPS组。结论: CCK-8可通过内源性H₂S介导的抗氧化、抑制PMN黏附聚集等效应发挥减轻LPS所致肺损伤的作用。     

过氧化物酶增殖体激活受体-β在大鼠急性肺损伤中的作用

目的探讨过氧化物酶增殖体激活受体-β（PPARβ-）在急性肺损伤（ALI）发生发展中可能的作用,以期从新的视角揭示ALI/ARDS的发病机理,并为ALI/ARDS的防治提供理论基础。方法采用颈静脉注射脂多糖（LPS）的方法复制大鼠ALI模型,随机分为对照组和LPS组。半定量RT-PCR方法测定肺组织中PPARβ-mRNA的表达水平;免疫组织化学染色法测定肺组织PPARβ-蛋白表达水平;同时测定动脉血气分析、肺组织湿/干（W/D）比值、肺组织MPO活性、光镜观察肺组织病理变化。结果LPS组大鼠PaO2显著降低（P〈0.01）,肺组织W/D比值、肺组织髓过氧化物酶（MPO）活性以及肺组织的病理积分均显著升高（P〈0.05）。免疫组化显示对照组大鼠肺组织可表达PPARβ-蛋白,主要位于肺泡上皮细胞及肺间质细胞;与对照组相比,LPS组大鼠肺组织PPARβ-蛋白的表达均显著升高（P〈0.05）,分布在肺泡上皮细胞及炎性细胞。RT-PCR结果显示对照组肺组织表达一定量的PPARβ-mRNA,与对照组比较,LPS组大鼠肺组织PPARβ-mRNA的表达量显著升高（P〈0.05）。结论正常大鼠肺泡上皮细胞及间质细胞存在PPARβ-,在ALI形成过程中PPARβ-蛋白和mRNA表达均显著增加,提示PPARβ-可能与ALI的炎症反应相关。     

黑木耳多糖对内毒素诱导急性肺损伤大鼠肺组织的保护作用

目的:探讨黑木耳多糖对LPS诱导急性肺损伤大鼠肺组织的保护作用及其机制。方法:将健康SD大鼠随机分为对照组、LPS组、地塞米松组以及黑木耳多糖低、中、高浓度组,根据分组,分别给予生理盐水或不同浓度黑木耳多糖预防性灌胃7 d,第8天腹腔注射生理盐水或LPS(8 mg/kg),地塞米松组在给予LPS后腹腔注射地塞米松(3 mg/kg)。造模12 h后于腹主动脉取血,并制肺组织匀浆和肺泡灌洗液。检测支气管肺泡灌洗液中蛋白含量、肺湿/干重比、髓过氧化物酶(MPO)、总抗氧化能力(T-AOC)、总超氧化物歧化酶(T-SOD)、一氧化氮合酶(NOS)、丙二醛(MDA)等指标,做组织切片HE染色并进行肺损伤评分。结果:应用黑木耳多糖干预后,急性肺损伤大鼠支气管肺泡灌洗液中蛋白含量明显下降,肺湿/干重比值降低;大鼠肺组织中MPO、NOS活性及MDA含量较LPS组降低,T-AOC含量和T-SOD活性较之升高;肺组织病理改变减轻,肺损伤指数下降。结论:黑木耳多糖具有保护LPS损伤大鼠肺组织的作用,其机制可能与其抗氧化作用有关。     

PI3K/Akt信号通路上调急性肺损伤大鼠肺泡上皮钠通道表达

目的 探讨PI3K/Akt信号通路对急性肺损伤(ALI)大鼠肺泡上皮钠通道(ENaC)α、β和γ亚基表达的影响.方法 成年SD大鼠随机分为对照组、ALI组(脂多糖)、胰岛素组及渥曼青霉素组,每组5只.观察肺组织病理改变,收集支气管肺泡灌洗液(BALF),测量总肺水含量,RT-PCR和Western blot测定ENaC mRNA和蛋白、p-Akt表达.结果 胰岛素组BALF蛋白含量、髓过氧化物酶(MPO)活性、总肺水含量较ALI组显著减少(P＜0.05),渥曼青霉素组BALF蛋白含量、MPO活性及总肺水含量较胰岛素组显著增加(P＜0.05).ALI组α-、β-和γ-ENaC蛋白表达显著低于对照组(0.33 ±0.06 vs 1.27 ±0.07,0.18±0.04 vs 0.72±0.04,0.37±0.04 vs0.69±0.05)(P＜0.05).胰岛素组蛋白表达α-ENaC(2.19 ±0.04)、β-ENaC(1.18 ±0.07)和γ-ENaC(1.18 ±0.08)显著高于ALI组(P＜0.05).渥曼青霉素组蛋白表达α-ENaC(0.86 ±0.09)、β-ENaC (0.58±0.05)和γ-ENaC (0.59±0.02)显著低于胰岛素组(P＜ 0.05).胰岛素组ENaC mRNA和p-Akt较ALI组显著升高(P＜0.05).渥曼青霉素组ENaC mRNA和p-Akt较胰岛素组显著降低(P＜0.05).结论 激活H3K/Akt通路上调3种ENaC亚基表达,从而清除肺水肿液.     

Azilsartan attenuates lipopolysaccharide-induced acute lung injury via the Nrf2/HO-1 signaling pathway

Acute lung injury (ALI) is a severe complication of sepsis and hemorrhagic shock with high morbidity. In the present study, the protective effect of Azilsartan on lipopolysaccharide (LPS)-induced ALI in mice was investigated to explore the potential therapeutic property of Azilsartan for the treatment of ALI. LPS was used to induce an ALI model in mice. Hematoxylin–eosin (HE) staining sections were then evaluated for the pathological state of lung tissues. Bronchoalveolar lavage fluid (BALF) protein concentration, wet/dry weight ratios of lung tissues, and pulmonary myeloperoxidase (MPO) activity were detected to determine the degree of pulmonary injury. The number of total cells, macrophages, and neutrophils in BALF were counted using a hemocytometer to illustrate the inflammatory cell infiltration. The lung function was monitored using a spirometer. The concentrations of interleukin-1β (IL-1β), monocyte chemoattractant protein-1 (MCP-1), and interleukin-8 (IL-8) were determined using enzyme-linked immunosorbent assay (ELISA). Oxidative stress was evaluated by the superoxide dismutase (SOD) activity, glutathione (GSH), and malondialdehyde (MDA) concentrations in the lung tissue. The expressions of nuclear erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were determined using Western blot analysis. Azilsartan therapy alleviated LPS-induced lung tissue damage, increased BALF protein concentration, lung wet to dry weight ratio, MPO activity, and macrophage and neutrophils infiltration. Also, Azilsartan ameliorated the production of inflammatory factors (IL-1β, MCP-1, and IL-8). Azilsartan ameliorated LPS-impaired lung SOD activity, the GSH concentration, and the MDA concentration. Mechanistically, Azilsartan activated the LPS-impaired Nrf2/HO-1 signaling pathway. Azilsartan therapy attenuates LPS-induced ALI via the Nrf2/HO-1 signaling pathway.

     

Formononetin Inhibited the Inflammation of LPS-Induced Acute Lung Injury in Mice Associated with Induction of PPAR Gamma Expression

Formononetin has shown a variety of pharmacologic properties including anti-inflammatory effect. In the present study, we analyzed the role of formononetin in acute lung injury induced by lipopolysaccharide (LPS) in mice. The cell counting in the bronchoalveolar lavage fluid (BALF) was measured. The animal lung edema degree was evaluated by wet/dry weight ratio. The superoxidase dismutase (SOD) activity and myeloperoxidase (MPO) activity was assayed by SOD and MPO kits, respectively. The levels of inflammatory mediators, tumor necrosis factor-α (TNF-α) and IL-6,were assayed by enzyme-linked immunosorbent assay method. Pathological changes of hung tissues were observed by HE staining. Peroxisome proliferator-activated receptor (PPAR)-γ gene expression was measured by real-time PCR. The data showed that treatment with the formononetin group markedly attenuated inflammatory cell numbers in the BALF, increased PPAR-γ gene expression and improved SOD activity and inhibited MPO activity. The histological changes of the lungs were also significantly improved by formononetin compared to LPS group. The results indicated that formononetin has a protective effect on LPS-induced acute lung injury in mice.     

氯沙坦抑制急性肺损伤大鼠肺组织LOX-1表达

 目的 研究氯沙坦干预大鼠急性肺损伤模型肺组织血凝素样氧化低密度血浆脂蛋白受体-1（LOX-1）表达的变化,探讨LOX-1在急性肺损伤中的表现及氯沙坦的干预机制。方法 将大鼠随机分成：对照组、脂多糖(LPS)组（5mg/kg）及氯沙坦治疗组（8mg/kg）,每组10只。检测动脉血氧分压（PaO2）、肺湿/干质量值（W/D）、支气管肺泡灌洗液(BALF)蛋白含量、血浆和BALF中TNF-α水平及肺组织病理变化,比色法测定肺组织髓过氧化物酶（MPO）活性,用RT-PCR法检测肺组织LOX-1 mRNA水平,western blot法检测肺组织中LOX-1、ICAM-1、Cleaved caspase-3蛋白的表达。结果 脂多糖处理后大鼠肺部炎症反应显著,PaO2较对照组下降,而肺W/D值、BALF蛋白含量及TNF-α水平、血浆TNF-α水平、MPO活性升高（P<0.05）;LOX-1 mRNA和LOX-1、ICAM-1及Cleaved caspase-3蛋白表达升高（P<0.05）。氯沙坦显著缓解上述变化（P<0.05）。结论 氯沙坦可能通过抑制LOX-1介导的炎症及细胞凋亡减轻急性肺损伤。     

骨髓来源间充质干细胞培养上清液减轻脂多糖诱导的小鼠急性肺损伤

 目的:观察经尾静脉输注骨髓间充质干细胞培养上清液（MSCs CdM）对脂多糖(LPS)诱导的小鼠急性肺损伤的治疗作用及其机制。方法:采用全骨髓培养法分离纯化骨髓间充质干细胞,传至第3代时观察细胞形态,流式细胞术检测细胞表面标志,并且收集上清液用超滤离心管进行离心。30只BALB/c小鼠随机分为对照组、 LPS模型组和MSCs CdM治疗组。对照组腹腔内注射生理盐水（0.01 mL/g）,LPS组和MSCs CdM治疗组腹腔内注射LPS（5 mg/kg,0.01 mL/g）制备急性肺损伤模型。造模1 h后经尾静脉输注MSCs CdM（MSCs  CdM治疗组）或生理盐水 （LPS组或对照组）300 μL。6 h后处死小鼠,留取标本检测肺组织病理形态学、肺组织湿干重比（W/D）、支气管肺泡灌洗液（BALF）中蛋白含量、血清及BALF中细胞因子水平和肺组织中髓过氧化物酶(MPO)的活性。结果:与对照组比较,LPS处理后肺组织病理损伤严重,BALF中蛋白、血清肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)含量、肺组织中MPO活性及肺组织湿干重比均显著升高。与LPS组比较,MSCs CdM治疗组肺组织病理损伤程度减轻,BALF中蛋白、血清TNF-α和IL-6含量、肺组织中MPO活性及肺组织湿干重比均显著降低,而BALF中白细胞介素10（IL-10）和角质细胞生长因子（KGF）水平显著高于LPS组和对照组。结论:骨髓间充质干细胞培养上清液可有效减轻LPS诱导的急性肺损伤,其作用机制可能与其调节肺部TNF-α、IL-6、IL-10和KGF的水平有关。     

The effects of the combination of sodium ferulate and oxymatrine on lipopolysaccharide-induced acute lung injury in mice

In our previous study, the remarkable analgesic and anti-inflammatory effects of the combination of sodium ferulate (SF) and oxymatrine (OMT) had been found. In this study, we investigated the effect of the combination of SF and OMT on acute lung injury using lipopolysaccharide (LPS)-induced acute lung injury (ALI) mouse model. The cell counting and the protein concentration in the bronchoalveolar lavage fluid (BALF) were measured. The animal lung edema degree was evaluated by wet/dry weight (W/D) ratio. The superoxidase dismutase (SOD) activity and myeloperoxidase (MPO) activity was assayed by SOD and MPO kits, respectively. The levels of inflammatory mediators including C-reactive protein (CRP) and tumor necrosis factor-α (TNF-α) were assayed by enzyme-linked immunosorbent assay method. The data showed that treatment with the combination of SF and OMT markedly attenuated inflammatory cell numbers and protein concentration in the BALF and improved SOD activity and inhibited MPO activity compared to LPS group. Moreover, the combination significantly inhibited the production of CRP and TNF-α in lung homogenate. The histological changes of the lungs were also more significantly improved by the combination. At the same dose, the obvious protective effect was not found in SF or OMT-treated alone group except that the protein concentration slightly decreased in SF group. The results indicated that the combination SF and OMT had a protective effect on LPS-induced ALI in mice, and the effect was much better than that of SF or OMT used alone.     

Adipolin/CTRP12对LPS致ARDS小鼠的保护作用及肺泡上皮钠离子通道的调节

目的:探讨adipolin/CTRP12对LPS致ARDS小鼠的作用及肺泡上皮钠离子通道(ENaC)的调节。方法:40只C57BL/6J小鼠按随机数字表分为对照组、LPS组、adipolin组和wortmannin(PI3K抑制剂)组,每组10只。苏木精-伊红(HE)染色观察肺组织病理改变;伊文氏蓝标记蛋白检测肺水清除率(AFC),BCA法测支气管肺泡灌洗液(BALF)中蛋白含量,评估肺组织通透性改变;ELISA检测BALF中白细胞介素(IL)-1β和肿瘤坏死因子(TNF)-α含量,髓过氧化物酶(MPO)试剂盒检测MPO活性,吉姆萨染色计数BALF总细胞数和多形核白细胞数,Western blot法检测肺组织α-ENaC蛋白表达和Akt磷酸化水平,实时荧光定量PCR检测肺组织α-ENaC的mRNA转录水平。结果:与control组相比,LPS组表现出典型的ARDS病理改变,肺损伤明显(P0.05),湿干重比(W/D)和BALF蛋白含量明显增高而AFC明显减弱(P0.05),BALF细胞计数、MPO活性、IL-1β含量和TNF-α含量明显升高(P0.05),而肺α-ENaC表达和Akt磷酸化水平显著下调(P0.05)。与LPS组相比,adipolin组肺损伤明显减轻(P0.05),W/D和BALF蛋白含量明显降低而AFC明显增强(P0.05),且BALF细胞计数、MPO活性、IL-1β含量和TNF-α含量均较LPS组显著降低(P0.05),伴α-ENaC表达和Akt磷酸化水平显著上调(P0.05)。而PI3K抑制剂wortmannin组与adipolin组相比,其肺损伤明显加重(P0.05),W/D和BALF蛋白含量明显增高,AFC明显减弱(P0.05),BALF细胞计数、MPO活性、IL-1β含量和TNF-α含量明显升高(P0.05),同时伴α-ENaC表达和Akt磷酸化水平显著下调(P0.05)。结论:Adipolin/CTRP12可通过PI3K/Akt信号通路介导的α-ENaC上调机制,增强肺水肿液清除能力,从而对LPS所致ARDS小鼠发挥保护性调控作用。     

Preventive Effects of Valnemulin on Lipopolysaccharide-Induced Acute Lung Injury in Mice

Valnemulin reportedly regulates inflammatory responses in addition to its in vitro antibacterial activity. In this study, we established a mouse model of lipopolysaccharide (LPS)-induced inflammatory lung injury and investigated the effect of valnemulin (100 mg/kg) on acute lung injury (ALI) 8 h after LPS challenge. We prepared bronchoalveolar lavage fluid (BALF) for measuring protein concentrations, cytokine levels, and superoxidase dismutase (SOD) activity, and collected lungs for assaying wet-to-dry weight (W/D) ratios, myeloperoxidase (MPO) activity, cytokine mRNA expression, and histological change. We found that the pre-administration of valnemulin significantly decreases the W/D ratio of lungs, protein concentrations, and the number of total cells, neutrophils, macrophages, and leukomonocytes, and histologic analysis indicates that valnemulin significantly attenuates tissue injury. Furthermore, valnemulin significantly increases LPS-induced SOD activity in BALF and decreases lung MPO activity as well. In addition, valnemulin also inhibits the production of tumor necrosis factor-α, interleukin-6, and interleukin-1β, which is consistent with mRNA expression in lung. The results showed that valnemulin had a protective effect on LPS-induced ALI in mice.