地塞米松对早期脓毒症大鼠急性肺损伤的保护作用研究

目的 观察地塞米松对早期脓毒症大鼠急性肺损伤(acute lung injury,ALI)的保护作用.方法 将雄性大鼠随机分为3组:生理盐水组(N组)、脂多糖组(L组)、脂多糖+地塞米松组(LD组).各组大鼠于造模后4h麻醉处死,行动脉血气分析,测定肺组织湿/干质量比值(W/D),用RT-PCR法检测肺组织水通道蛋白1(aquaporin 1,AQP1 )mRNA表达水平.结果 与L组比较,LD组大鼠肺组织湿/千质量比明显降低,AQP1 mRNA表达水平升高,动脉血气改善.结论 地塞米松对早期脓毒症大鼠ALI有保护作用,该作用与上调肺AQP1 mRNA表达水平有关.     

瑞芬太尼对脓毒症大鼠急性肺损伤保护作用的实验研究

目的观察瑞芬太尼对脓毒症所致急性肺损伤(ALI)大鼠肺湿/干质量比、肺组织病理学改变及炎症因子及丙二醛(MDA)和超氧化物歧化酶(SOD)的影响,为瑞芬太尼治疗脂多糖(LPS)诱导的ALI提供理论依据。方法采用雄性清洁级Wistar大鼠32只,体质量270～320g,随机分为四组:对照组、ALI组、瑞芬太尼处理组和瑞芬太尼对照组。ALI组于20min内尾静脉注射LPS 15mg/kg制作脓毒症ALI模型,对照组用相同量的生理盐水代替;瑞芬太尼对照组和瑞芬太尼处理组,先注射等量生理盐水/LPS,后给予瑞芬太尼(0.04mg/kg)尾静脉泵入40min。测定各组大鼠造模后6h后的肺湿/干比;苏木精-伊红(HE)染色方法对肺组织进行病理观察,ELISA检测肿瘤坏死因子(TNF-α),白介素-6(IL-6)的水平,并测定SOD活性及MDA的含量。结果与对照组比较,ALI组大鼠肺干湿比、肺组织TNF-α、IL-6水平、MDA含量增加而SOD活性显著降低(P 0.05);而瑞芬太尼显著改善上述指标的变化。肺组织病理学观察发现ALI大鼠肺组织结构损伤严重,可见大量炎性细胞聚集。而瑞芬太尼处理组大鼠的上述病理学改变则明显减轻。结论瑞芬太尼可以通过抑制肺组织炎症因子的释放,减轻氧化应激途径保护LPS所致的急性肺损伤。     

炎调方对脓毒症急性肺损伤大鼠肺组织髓过氧化物酶和丙二醛水平的影响

目的 观察炎调方对脓毒症急性肺损伤(ALI)大鼠肺组织髓过氧化物酶(MPO)、丙二醛(MDA)水平的影响。方法 清洁级健康雄性SD大鼠随机分为对照组、假手术组、模型组、炎调方组、地塞米松组,炎调方组ig 9.9 g/kg炎调方,地塞米松组ig 0.45 mg/kg地塞米松,每天给药1次,连续给药3 d。末次ig 2 h后采用盲肠结扎穿孔术(CLP)制备脓毒症ALI模型,分别于造模后24 h处死动物,进行肺组织HE染色,测定各组大鼠湿/干重比(W/D),肺组织MPO、MDA水平。结果 模型组大鼠肺组织损伤程度、MPO及MDA水平较对照组及假手术组显著增高(P<0.01),炎调方组、地塞米松组肺组织损伤程度、MPO及MDA水平较模型组显著降低(P<0.05、0.01)。结论 炎调方可有效减轻脓毒症ALI大鼠的炎症及氧化应激反应。     

低氧诱导因子-1α对脓毒症肠黏膜屏障的保护作用

目的:探讨低氧诱导因子-1α(HIF-1α)对脓毒症肠黏膜屏障的影响及机制。方法:SD大鼠随机分4组:假手术组、脓毒症组、脓毒症+HIF-1α刺激剂组(脓毒症+DMOG组)、脓毒症+HIF-1α抑制剂组(脓毒症+Bay87-2243组),每组6只。采用盲肠结扎穿孔(cecal ligation and perforation,CLP)建立脓毒症模型。ELISA检测大鼠血浆炎性标记物IL-1β、IL-6、TNF-α,氧化应激标记物丙二醛(MDA)以及抗氧化因子超氧化物歧化酶(SOD)、过氧化氢酶(CAT)的水平。Western blot检测大鼠肠黏膜HIF-1α表达。HE染色检测肠黏膜病理学损伤。结果:与假手术组相比,脓毒症大鼠炎症因子、氧化应激因子以及HIF-1α显著上调(P<0.05);给予腹腔注射DMOG明显降低脓毒症大鼠血浆IL-1β、IL-6、TNF-α、MDA水平(P<0.05);增加血浆SOD、CAT水平(P<0.05);HIF-1α表达上调(P<0.05),肠黏膜病理损伤减轻,Chiu's评分显著降低(P<0.05)。口服灌胃Bay87-2243则得到相反的结果。结论:HIF-1α对脓毒症肠黏膜损伤具有保护作用,其机制可能与缓解脓毒症炎性反应和抑制氧化应激水平有关。     

连续肾脏替代治疗在脓毒性休克中的应用

<正>脓毒症(sepsis)指的是由感染引起的全身炎性反应。严重脓毒症(severe sepsis)和脓毒性休克(septic shock)即全身性感染伴有器官功能障碍,组织低灌注或低血压(收缩压<90mmHg,或下降幅度超过40mmHg)。脓毒症是危重患者主要的死亡原因之一,很容易发展为休     

内毒素致急性肺损伤大鼠水通道蛋白1、5表达的影响

目的:观察细菌内毒素(LPS)致急性肺损伤(ALI)大鼠肺组织的变化,探讨大鼠急性肺损伤模型的复制.方法:SPF级雄性SD大鼠42只,随机分为ALI组和正常对照组(NS组),每组21只.ALI组子LPS(4 mg·kg-1,浓度4 mg·ml-1)舌下静脉注射;NS组予生理盐水0.2ml舌下静注,动态观察注射后2,6,12 h肺组织湿干质量(W/D)、肺组织病理学改变、支气管肺泡灌洗液(BALF)的细胞总数及中性粒细胞百分比、免疫组化与Western blot法肺组织AQP1蛋白表达的变化.结果:与NS组相比,ALI组大鼠W/D值、BALF细胞总数、中性粒细胞百分比、免疫组化与Western blot法肺组织AQP1、5蛋白表达的变化于LPS静脉注射后2h开始升高,第6 h达高峰.ALI组与NS组各时相点间的差异均具有统计学意义(P＜0.05).结论:静脉注射LPS可导致大鼠ALI,肺炎症损伤反应的高峰于LPS致伤后6～12h最为明显.     

脓毒症中线粒体DNA 的研究进展

脓毒症是重症监护病房常见急危重症,发病率、病死率均较高,早期识别困难,临床医师若不能在早期及时识别及救治脓毒症患者,可进一步发展为脓毒症休克、多脏器功能 衰竭 ,从而增加病死率。近年来,有研究表明线粒体损伤与脓毒症发生发展密切相关,而线粒体DNA(mtDNA)损伤是线粒体损伤重要机制之一。“危险模型”提出,当细胞受到损害时,无论刺激的性质和强度,细胞均会将其组分释放到细胞外空间,驱动免疫或炎症反应,这类内源性分子被称为DAMP。mtDNA 作为一种DAMP,在脓毒症中可能通过Toll 样受体9(TLR9)和炎性小体影响机体炎症反应,为mtDNA 作为脓毒症诊断的新生物学标记提供了可能。脓毒症患者体内游离mtDNA 异常升高可能与患者病死率显著相关,本文就近年来脓毒症中mtDNA 研究进展作一综述     

通腑泻肺法对ALI/ARDS大鼠血清炎症因子表达的影响

目的观察通腑泻肺法对ALI/ARDS大鼠血清炎症因子的影响,探讨ALI/ARDS肺肠同治的生物学基础。方法将100只健康Wistar大鼠随机分为5组:空白对照组、模型组、治肺组、治肠组和肺肠同治组,采用尾静脉注射油酸加内毒素方法造成大鼠ALI/ARDS模型,观察大鼠血清TNF-α、IL-1β、IL-6、IL-10含量。结果通腑泻肺法可以有效降低大鼠全身炎性因子TNF-α、IL-1β、IL-6、IL-10表达水平。结论 (1)通腑泻肺法能够明显减轻ALI/ARDS大鼠免疫炎症损伤,其保护作用较单纯用泻肺法或通腑法效果更有优势;(2)泻肺法、通腑法和通腑泻肺法对ALI/ARDS大鼠血清炎性因子作用靶点不同。     

热休克蛋白70在大鼠急性肺损伤中的表达

目的:探讨急性肺损伤(ALI)大鼠肺组织热休克蛋白70(HSP70)的表达。方法:雌性Wistar大鼠36只,随机分为两组:急性肺损伤组(A组)、对照组(B组),大鼠尾静脉注射脂多糖(LPS)复制ALI模型。所有的动物于注射LPS后2,4,6 h处死,测定肺组织HSP70的表达、超氧化物歧化酶(SOD)活性及丙二醛(MDA)的含量。结果:A组同B组相比,各时间点肺组织HSP70表达均增多,以2,6 h升高明显(P<0.01),MDA含量增高(P<0.05),SOD活性明显降低(P<0.05)。结论:HSP70在ALI后的表达增多可能与其参与LPS所致肺损伤后的组织保护有关。     

重组人生长激素对败血症大鼠急性肺损伤的治疗及其机制

目的　观察重组人生长激素(rhGH)对败血症大鼠急性肺损伤(ALI)的治疗效果并探讨其作用机制。方法　采用鸵鸟株E .coli复制败血症大鼠ALI模型。观测对照组、ALI组及rhGH治疗组大鼠的肺组织病理评分、支气管肺泡灌洗液(BALF)中NO水平及肺组织诱生型一氧化氮合酶(iNOS)蛋白的表达情况。结果　ALI组大鼠肺组织病理评分、BALF中NO水平及肺组织iNOS蛋白表达水平明显高于对照组,且NO与iNOS蛋白表达水平呈明显正相关。rhGH能明显降低败血症大鼠BALF中NO及肺组织iNOS水平,减轻肺损伤程度。结论　由iNOS诱导产生的过量NO可促进败血症大鼠ALI的发生;rhGH对败血症大鼠ALI具有较理想的治疗效果,其机制可能与rhGH通过影响iNOS表达来降低肺组织NO水平等有关。     

己酮可可碱弱化大鼠脓毒症急性肺损伤对p38MAPK/IκBα磷酸化的影响

目的探讨己酮可可碱（pentoxifylline,PTX）对腹腔感染致脓毒症急性肺损伤发挥保护作用的上游信号机制。方法采用盲肠结扎穿孔（cecal ligation and puncture,CLP）致脓毒症模型,将大鼠随机分为假手术组、脓毒症组、脓毒症＋西黄著胶组、脓毒症＋生理盐水组、脓毒症＋SB203580组、脓毒症＋PTX组。检测假手术组、脓毒症组各时间点（1,3,6,12,24 h）p38MAPK及IκBα的磷酸化,然后选择1,6,24 h组分别检测应用SB203580或PTX后p38MAPK及IκBα的表达,同时检测血浆TNF-α、IL-6的含量并观察24 h组肺组织病理改变。结果与假手术组比较,脓毒症组在各个时间点p38MAPK及IκBα均有较强的表达,SB203580或PTX预处理后各组的p38MAPK及IκBα的磷酸化明显受到抑制,且与血浆TNF-α、IL-6的含量以及肺的病理切片变化一致。结论 PTX对脓毒症急性肺损伤的保护作用以及抑制促炎因子产生的作用可能是通过抑制p38MAPK的磷酸化从而抑制NF-κB的活化而产生的。     

己酮可可碱对大鼠脓毒症急性肺损伤p38MAPK活性的影响

目的探讨己酮可可碱（pentoxifylline,PTX）对腹腔感染致脓毒症急性肺损伤发挥肺保护作用与p38MAPK活化的关系。方法采用盲肠结扎穿孔致脓毒症模型,将大鼠随机分为Ⅰ组（Sham组）、Ⅱ组（脓毒症CLP组）、Ⅲ组（脓毒症加西黄著胶CLP＋V组）、Ⅳ组（脓毒症加生理盐水CLP＋N组）、Ⅴ组（脓毒症加SB203580 CLP＋SB组）、Ⅵ组（脓毒症加己酮可可碱CLP＋PTX组）,其中Ⅲ组、Ⅳ组为溶媒对照组。用Western Blot检测假手术组,脓毒症1,3,6,12,24 h后p38MAPK的磷酸化,然后选择1,6,24 h分别检测应用SB203580或PTX后p38MAPK的表达,同时检测血浆TNF-α、IL-6的含量并观察24 h内肺组织病理改变。结果与假手术组比较,脓毒症组在各个时间点p38MAPK均有较强的表达,SB203580或PTX预处理后各组的p38MAPK的磷酸化明显受到抑制,且与血浆TNF-α、IL-6的含量以及肺的病理切片变化一致。结论己酮可可碱可能是通过抑制p38MAPK的磷酸化抑制促炎因子的过度表达,发挥对脓毒症急性肺损伤的保护作用。     

Exogenous administration of mitochondrial DNA promotes ischemia reperfusion injury via TLR9-p38 MAPK pathway

Previous studies have shown a role of mitochondrial DNA (mtDNA) in innate immunity. However, the specific role of mtDNA in acute myocardial infarction remains elusive. This study was designed to examine the damaging effect of mtDNA on cardiomyocytes. H9c2s cells were incubated with purified mtDNA or nuclear DNA with or without pretreatment by chloroquine, an inhibitor of Toll-like receptor 9(TLR9). The cell viability was tested by MTT. To demonstrate the toxicity of mtDNA, mtDNA fragments were injected into rats 10 min before ischemia for 30 min and reperfusion for 24 h. Infarct size was measured by TTC staining. Apoptosis of myocardium was detected by TUNEL staining and caspase-3 activity. The levels of TLR9, p-p38 MAPK, and p38 MAPK were detected by western blotting. The results showed that exogenous mtDNA reduced the viability of H9c2s cells and induced TLR9 expression, caspase 3 activation and p38 mitogen-activated protein kinase (MAPK) phosphorylation. However, these effects were inhibited by chloroquine. In contrast, nuclear DNA did not have these effects. Intravenous injection of mtDNA into rats aggravated ischemia-reperfusion injury and increased infarction area through TLR9-p38 MAPK activation. We concluded that mtDNA released into the circulation by AMI may has detrimental effect on myocardium through aggravating ischemia-reperfusion injury via TLR9-p38 MAPK pathway.     

地塞米松抑制哮喘大鼠肺组织p38蛋白激酶的表达

目的:探讨支气管哮喘大鼠肺组织p38蛋白激酶(p38 MAPK)表达的变化以及地塞米松对其影响.方法:复制大鼠哮喘模型,随机分成3组:正常对照组、哮喘对照组和地塞米松(DEX)干预组.分别采用酶联免疫吸附法(ELISA)和蛋白质印迹检测支气管肺泡灌洗液(BALF)IL-5含量和肺组织磷酸化p38 MAPK表达的变化,并观察气道阻力、BALF中EOS计数以及肺组织病理学变化.结果:哮喘对照组大鼠肺组织磷酸化p38 MAPK表达水平及气道阻力、BALF中IL-5含量和EOS计数均较正常对照组显著增加(P＜0.01);DEX干预组上述指标较哮喘对照组显著降低(P＜0.01),肺组织病理学损伤程度明显减轻.肺组织磷酸化p38 MAPK表达水平与气道阻力、BALF中IL-5含量和EOS计数之间分别呈显著正相关(r=0.77、0.63、0.65,P＜0.01).结论:p38 MAPK可能参与了支气管哮喘的发病过程.DEX对哮喘的治疗作用至少部分与抑制磷酸化p38 MAPK的表达有关.     

炎调方对脓毒症急性肺损伤大鼠肺组织热休克蛋白70 mRNA和p38丝裂原活化蛋白激酶的影响

目的 探讨炎调方对脓毒症急性肺损伤（ALI）大鼠肺组织热休克蛋白70（HSP70）mRNA和p38丝裂原活化蛋白激酶（p38 MAPK）的调控作用。方法 将清洁级健康雄性SD大鼠随机分为假手术组、模型组、炎调方（生药量9.9 g/kg）组、谷氨酰胺组、槲皮素组,采用盲肠结扎穿孔术法（CLP）建立脓毒症ALI模型。观察大鼠一般状况,各组分别于术后4、6、8、10、12、18、24 h采集肺组织标本,HE染色观察肺组织病理学改变;实时荧光定量PCR法检测肺组织HSP70 mRNA的表达水平;Western Blotting法检测肺组织p38 MAPK的蛋白表达水平。结果 炎调方组、谷氨酰胺组大鼠与模型组比较,术后不同时间点的精神状态、活动量等明显好转,可见少量稀便;肺组织损伤程度明显减轻,腔内出血较少,肺实变较轻。炎调方组不同时间点的肺组织HSP70 mRNA表达水平显著高于假手术组、模型组（除24 h时间点外）、槲皮素组（P<0.01）,在8、12、18、24 h时间点显著低于谷氨酰胺组（P<0.01）;炎调方组不同时间点的肺组织p38 MAPK的蛋白表达量显著低于模型组、槲皮素组（P<0.01）,在4、6、10 h时间点明显低于谷氨酰胺组（P<0.05、0.01）。结论 炎调方可通过上调肺组织HSP70 mRNA的表达,下调肺组织p38 MAPK的蛋白表达,改善肺组织损伤,对脓毒症ALI发挥保护作用。     

Hyaluronan ameliorates LPS-induced acute lung injury in mice via Toll-like receptor (TLR) 4-dependent signaling pathways

Toll-like receptor-4 (TLR4) signaling has been implicated in innate immunity and acute inflammation following acute lung injury (ALI). As such, modulating inflammatory response through TLR4 represents an attractive therapeutic approach to treat ALI. Increasing evidence demonstrates that hyaluronan (HA) can modulate TLR4 activation and has shown early promise as a therapeutic agent in ALI. However, the mechanism associated with HA has not been fully elucidated. In the current study, we sought to determine the effects of HA on lipopolysaccharide (LPS)-induced inflammatory response and gain insights into the mechanism of action in mice with intratracheal instillation of LPS. Our results demonstrate that in contrast to mice challenged with LPS, pretreatment with HA significantly inhibited inflammatory cell recruitment, attenuated lung injury and suppressed the level of cytokine/chemokine in bronchial alveolar lavage fluid (BALF). Investigation of the mechanism responsible for inhibition of LPS activation showed HA treatment significantly inhibited the nuclear translocation of NF-κB p65 and protein expression of myeloid differentiation primary response protein (MyD88) and TIR-domain-containing adapter-inducing interferon-β (TRIF) and p38 MAPK, JNK and ERK activation in lung tissue. Furthermore, we compared the protection effect of HA in TLR4-deficient mice with those of genetically matched wild type (WT) mice in an acute model of lung injury. However, in TLR4-deficient mice, HA pretreatment before LPS instillation fail to affect the LPS response. Therefore, our findings suggest that HA pretreatment attenuated LPS-induced ALI and the anti-inflammatory function of HA was partial dependent on TLR4, which shed new light on potential elements that regulate the lung injury response.     

Apocynin ameliorates endotoxin-induced acute lung injury in rats

Acute lung injury (ALI) is a serious clinical syndrome with a high rate of mortality. In this study, the effects of apocynin, a NADPH-oxidase (NOX) inhibitor on lipopolysaccharide (LPS)-induced ALI in rats were investigated. Male Sprague–Dawley rats were treated with apocynin (10 mg/kg) intraperitoneally (i.p.) 1 h before LPS injection (10 mg/kg, i.p.). The results revealed that apocynin attenuated LPS-induced ALI as it decreased total protein content, lactate dehydrogenase (LDH) activity and the accumulation of the inflammatory cells in the bronchoalveolar lavage fluid (BALF), In addition, apocynin significantly increased superoxide dismutase (SOD) and reduced glutathione (GSH) activities with significant decrease in the lung malondialdehyde (MDA) content as compared to LPS group in lung tissue and decreased pulmonary artery contraction induced by LPS. It also upregulated mRNA expression of inhibitory protein kappaB-alpha (NFκBia) and downregulated mRNA expression of Toll-Like receptor 4 (TLR4) and decreased inflammation observed in lung tissues.Collectively, these results demonstrate the protective effects of apocynin against the LPS-induced ALI in rats through its antioxidant and antiinflammatory effect that may be attributed to the decrease in mRNA expression of TLR4 and increasing that of NFκBia.     

辛伐他汀对脓毒症小鼠肺损伤的保护作用

目的观察辛伐他丁对脓毒症小鼠急性肺损伤的保护作用,并探讨其可能机制。方法采用盲肠结扎穿孔术制备脓毒症小鼠模型,将72只雄性C57BL/6小鼠随机分成3组:假手术组、脓毒症急性肺损伤组(脓毒症组)、脓毒症+辛伐他汀治疗组(治疗组)。治疗组给予辛伐他汀0.2μg/g,q12h腹腔注射1周;假手术组、脓毒症组给予等量安慰剂腹腔注射1周。分别于造模后6、12、24 h留取肺脏标本。HE染色观察肺组织病理学变化,免疫组化检测肺组织toll样受体4(Toll-like receptor 4,TLR4)蛋白的表达,ELISA测定肺组织匀浆中IL-1β及TNF-α的表达水平。结果与假手术组比较,造模后6、12、24 h,脓毒症组肺组织病理学评分、肺组织TLR4蛋白的表达,以及TNF-α、IL-1β水平明显升高(P<0.05);与脓毒症组相比,治疗组上述指标明显降低(P<0.05)。结论辛伐他汀通过抑制TLR4信号转导通路,减少其下游炎症介质TNF-α、IL-1β的释放,对脓毒症导致的急性肺损伤具有一定保护作用。     

FTY-720通过TGF-β1/P38 MAPK/NF-κB信号通路对小鼠肺纤维化模型的影响

目的探讨FTY-720对博来霉素诱导的小鼠肺纤维化的影响及其作用机制。方法将40只无特定病原体级健康雄性Balb/c小鼠随机分为对照组、博来霉素组、生理盐水+FTY-720对照组、博来霉素+FTY-720组,每组各5只,在7天、14天处死小鼠,共8组。应用HE染色和Masson染色观察肺组织炎症及肺纤维化情况;通过瑞士-吉姆萨染色计数支气管肺泡灌洗液细胞;BCA法测定BALF蛋白含量;ELISA方法检测BALF上清液中炎症因子IL-1β、TNF-α的表达水平;免疫组织化学法检测肺组织中COL1A1的表达水平;Western blot方法检测TGF-β1、p-P38 MAPK和NF-κB表达水平的变化。结果FTY-720显著降低BLM诱导增加的小鼠肺组织细胞外胶原沉积;减少小鼠肺组织BALF中IL-1β和TNF-α表达水平,和蛋白质含量、细胞数;FTY-720抑制TGF-β1活性与P38 MAPK的磷酸化,进而抑制NF-κB的表达与活化。结论FTY-720通过抑制TGF-β1/P38 MAPK/NF-κB信号通路,进而抑制博来霉素诱导小鼠的肺纤维化。