Toll样受体4在失血性休克复苏致小鼠急性肺损伤中的作用

目的 探讨Toll样受体4(TLR4)在失血性休克复苏致小鼠急性肺损伤中的作用.方法 TLR4基因突变型C3H/HeJ小鼠和野生型C3H/HeN小鼠各24只,两种品系小鼠各随机分为2组:假手术组(S组,n=6)、失血性休克复苏组(HSR组,n=18)制备失血性休克复苏模型,并于复苏后6、24、48 h时各取6只小鼠颈动脉放血处死后开胸取肺组织,免疫组织化学法检测p38 MAPK表达水平,酶联免疫吸附法测定白细胞介素(IL)-10和IL-6含量,透射电镜下观察肺组织超微结构.结果 与C3H/HeN小鼠比较,C3H/HeJ小鼠复苏后肺组织p38 MAPK表达下调,IL-6和IL-10含量降低(P<0.05或0.01),病理损伤程度减轻.两种品系小鼠中,与S组比较,HSR组复苏后24 h时肺组织IL-6和IL-10含量增加,复苏后48 h时肺组织IL-6含量增加(P<0.05或0.01);C3H/HeN小鼠中,与S组比较,HSR组复苏后6 h和24 h时肺组织p38 MAPK蛋白表达上调,复苏后48 h时肺组织IL-10含量增加(P<0.01).结论 TLR4参与小鼠失血性休克复苏致急性肺损伤的发生,其机制与激活p38 MAPK信号转导通路有关.     

Toll样受体4对失血性休克小鼠肺组织血红素加氧酶和诱导型一氧化氮合酶的影响

目的 观察Toll样受体4(TLR4)对失血性休克小鼠所致急性肺损伤(ALI)中肺组织血红素加氧酶-1(HO-1)和诱导型一氧化氮合酶(iNOS)的影响.方法 TLR4基因突变型小鼠C3H/HeJ和野生型小鼠C3H/HeN(48只),随机分为假手术组和失血性休克(6、24、48 h)组.采用小鼠失血性休克致急性肺损伤模型,观察血气分析,HO-1和iNOS蛋白表达,白细胞介素-6(IL-6)水平,肺组织肺湿/干重比和病理形态学改变.结果 与假手术组比较,C3H/HeN和C3H/HeJ小鼠失血休克后24 h肺组织HO-1蛋白强阳性表达,IL-6含量明显增加为365.38±48.26和300.89±39.34;6 h肺组织iNOS蛋白强阳性表达(P＜0.01).与C3H/HeN小鼠比较,C3H/HeJ小鼠失血休克后24 h肺组织HO-1、IL-6含量和W/D明显降低(P＜0.05);6 h肺组织iNOS蛋白显著减少为0.049±0.013.病理学检查显示失血休克后各时点肺组织损伤程度较假手术组明显加重.结论 TLR4在失血性休克后ALI过程中被激活,通过影响HO-1和iNOS的表达参与了失血性休克致急性肺损伤的过程.     

P38MAPK信号通路在失血性休克复苏诱发急性肺损伤小鼠HO-1表达上调中的作用

目的 探讨p38分裂原激活蛋白激酶(p38MAPK)信号通路在失血性休克复苏诱发急性肺损伤小鼠血红素加氧酶1(HO-1)表达上调中的作用.方法 SPF级野生型小鼠C3H/HeN32只32只,10～12周龄,体重20～25 g,随机分为4组(n=8),假手术组(S组):只进行手术操作;失血性休克复苏组(HSR组):股动脉放血,至MAP为40 mm Hg,通过放血和回输血液维持MAP 35～45mmHg,60 min后回输全部血液和等失血量的乳酸钠林格氏液复苏;FR167653组(FR组):静脉注射p38MAPK抑制剂FR167653 5 mg/kg;FR+HSR组:于放血前30 min静脉注射FR167653 5 mg/kg.复苏后6 h处死小鼠,取肺组织,观察病理学结果,并进行病理学评分,计算肺湿/干重比,检测肺组织髓过氧化物酶(MPO)、IL-10、IL-6和HO-1水平以及p38MAPK的激活水平.结果 与S组比较,HSR组肺组织病理学评分、肺湿/干重比、MPO、IL-6、IL-10、HO-1和p38MAPK的激活水平升高,HSR+FR组肺组织病理学评分、肺湿/干重比和HO-1表达水平升高(P＜0.01),FR组上述指标差异无统计学意义(P＞0.05);与HSR组比较,HSR+FR组肺组织病理学评分、肺湿/干重比、MPO、IL-6、IL-10、HO-1和p38 MAPK激活水平降低(P＜0.01).结论 p38MAPK信号通路介导了失血性休克复苏诱发急性肺损伤小鼠HO-1的表达上调.     

富氢液对脂多糖致小鼠急性肺损伤的影响

目的 评价富氢液对脂多糖(LPS)致小鼠急性肺损伤的影响.方法 成年雄性C57BL/6小鼠32只,体重20 ～ 25 g,采用随机数字表法,将其分为4组(n=8):对照组(C组)、富氢液组(H2组)、急性肺损伤组(ALI组)和急性肺损伤+富氢液组(ALI+ H2组).ALI组和ALI+ H2组分别雾化吸入LPS25 μg(溶于PBS中)制备ALI模型,C组和H2组分别雾化吸入无菌PBS 50μl.H2组和ALI+H2组雾化吸入PBS或LPS后1和12 h时腹腔注射0.6 mmol/L富氢液5 ml/kg.LPS或PBS处理后24 h时,机械通气15 min,行动脉血气分析,计算氧合指数.然后收集支气管肺泡灌洗液(BALF),测定蛋白浓度,计数中性粒细胞(PMN).采用ELISA法检测BALF中TNF-α、IL-1β、IL-6和高迁移率族蛋白1(HMGB1)的浓度.然后处死小鼠,取肺组织,行病理学损伤评分,测定湿重/干重(W/D)比、髓过氧化物酶(MPO)和caspase-3的活性,计算细胞凋亡指数(AI).结果 与C组比较,H2组氧合指数、BALF总蛋白、TNF-α、IL-1β、IL-6和HMGB1的浓度和PMN计数、肺组织病理学损伤评分、W/D比、MPO和caspase-3的活性以及AI差异均无统计学意义(P＞0.05),ALI组和ALI+H2组氧合指数降低,BALF蛋白、TNF-α、IL-1β、IL-6和HMGB1的浓度、PMN计数、肺组织病理学损伤评分、W/D比、MPO和caspase-3 的活性以及AI均升高(P＜0.05);与ALI组比较,ALI+ H2组氧合指数升高,BALF总蛋白、TNF-α、IL-1β、IL-6和HMGB1的浓度、PMN计数、肺组织病理学损伤评分、W/D比、MPO和caspase-3的活性以及AI均降低(P＜0.05).结论 富氢液可减轻LPS致小鼠急性肺损伤,可能与其抗炎和抗凋亡作用有关.     

急性肺损伤小鼠外周血树突状细胞数量及功能状态的变化

目的 探讨急性肺损伤(ALI)小鼠外周血树突状细胞(DC)数量及功能状态的变化.方法 雄性SPF级C57BL/6小鼠36只,6～8周龄,体重18～22 g,采用随机数字表法,将小鼠分为2组(n=18):对照组(C组)和ALI组.ALI组气管内注射脂多糖2 mg/kg,以制备ALI模型,C组注入等容量的PBS.于注入LPS或PBS后6、12和24 h(T1-T3)时随机取6只小鼠,麻醉后开胸取右心室血样,采用流式细胞仪检测外周血DC数量及DC表达CD80、MHCⅡ的水平;随后处死小鼠,取肺组织,计算湿重/体重(WW/BW)比,光镜下观察病理学结果,行肺损伤评分,采用ELISA法检测肺组织IL-6含量.结果ALI组可见肺泡间隔增宽、充血、出血及大量炎性细胞浸润等病理改变.与C组比较,ALI组各时点肺组织WW/BW比、IL-6含量、肺损伤评分升高,T1时外周血DC数量减少,T3时增多,T2、T3时DC表达MHCⅡ水平上调(P＜0.05).与T1时比较,ALI组T2时肺组织IL-6含量升高,T2、T3时外周血DC数量增多,DC表达MHCⅡ水平上调(P＜0.05);与T2时比较,ALI组T3时外周血DC数量增多(P＜0.05);2组各时点外周血DC表达CD80水平差异无统计学意义(P＞0.05).结论 ALI小鼠外周血DC数量早期减少,随后逐渐增多,功能呈渐成熟状态.     

内质网IRE1α/XBP1信号通路在小鼠内毒素性急性肺损伤中的作用：与NLRP3炎症小体的关系

目的评价内质网肌醇需要酶1α/X盒结合蛋白1(IRE1α/XBP1)信号通路在小鼠内毒素性急性肺损伤(ALI)中的作用及其与NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体的关系。方法 SPF级健康雄性C57BL/6小鼠36只, 6～8周龄, 体质量25～30 g, 采用随机数字表法分为3组(n=12):对照组(C组)、内毒素性ALI组(ALI组)和内毒素性ALI+STF-083010组(ST组)。ALI组和ST组雾化吸入LPS 3 mg/ml 30 min制备小鼠内毒素性ALI模型, C组雾化吸入等量生理盐水, ST组于雾化吸入LPS前1 h时腹腔注射IRE1α/XBP1信号通路阻断剂STF-083010 50 mg/kg, 其余2组腹腔注射等容量生理盐水。LPS或生理盐水雾化吸入后24 h时处死小鼠, 收集支气管肺泡灌洗液(BALF)并取肺组织标本, 肺组织HE染色后光镜观察病理学结果, 行肺损伤评分并计算肺湿重/干重(W/D)比值;ELISA法测定BALF上清液IL-1β和IL-18浓度;Western blot法检测肺组织p-IRE1α、XBP1s、NLRP3、ASC和...     

Effect of TLR-4 and HO-1 on acute lung injury induced by hemorrhagic shock in mice

Objective： To examine whether TLR-4 has an ettect on hemorrhage induced changes in lung, and to investigate the change of heme oxygenase-1 （HO-1） on acute lung injury （ALl） induced by hemorrhagic shock in mice.
Methods： Forty-eight male mice, including C3H/HeN mice and C3H/HeJ mice, were randomly divided into sham group （n=12）, hemorrhagic shock group with twelve mice in each phase. Blood pressure （BP） was monitored continuously by attaching carotid artery catheter to a strain gauge pressure transducer/ polygraph. Arterial blood samples were taken for blood gas analysis. A mouse model of non-lethal hemorrhagic shock and resuscitation was used to observe pulmonary myeloperoxidase （MPO） activity and wet/dry weight ratio （W/D）. The expression of HO- 1 was observed by means of RT-PCR and immunohistochemistry. IL-6 and IL-10 in lung tissue homogenate were assayed by enzyme-linked immunosorbent assay （ELISA）. The pulmo- nary pathologic changes were observed under electron microscope and light microscope.
Results： Compared with sham group, the expression of HO- 1 in lung tissue was significantly higher in Hem 24 h and Hem 48 h of C3H/HeN mice （P〈0.01）. The expression of HO-1 mRNA and the levels of IL-6, IL-10 and MPO in lung tissue were markedly increased in Hem 24 h （P〈0.01 or P〈0.05）; Compared with C3H/HeN mice, the expression of HO- 1 rnRNA and the levels of IL-6 and IL-10 in C3H/HeJ mice significantly decreased in Hem 24 h and Hem 48 h （P〈0.01 or P〈O.05）, and the W/D, MPO in C3H/HeJ mice were obvi- ously lower in Hem 24 h （P〈0.05）. The injuries of lung tissues after hemorrhagic shock have been demonstrated by histological examination with electron microscope and light microscope.
Conclusions： TLR-4 and HO-1 might modulate the bal- ance of pro- and anti-inflammatory processes in inflamma- tory reaction of hemorrhagic shock-induced ALl, and the activation of Toll-like receptor might induce the transcrip- tion activity of HO- 1, which may play a k     

PIAS调控PPARγ的SUMO化修饰在小鼠内毒素性急性肺损伤内源性保护机制中的作用

目的评价小泛素相关修饰物(SUMO) E3连接酶(PIAS)调控过氧化物酶体增殖物激活受体γ(PPARγ)的SUMO化修饰在小鼠内毒素性急性肺损伤(ALI)内源性保护机制中的作用。方法实验Ⅰ清洁级野生型雄性C57BL/6小鼠24只, 6～8周龄, 体质量18～22 g, 采用随机数字表法分为4组(n=6):对照组(C组)、ALI组、ALI+ PPARγ诱导剂TZD组(ALI+T组)、ALI+TZD+SUMO化抑制剂漆树酸组(ALI+T+A组)。尾静脉注射LPS 15 mg/kg制备内毒素性ALI模型。ALI+T+A组注射LPS前1 h时腹腔注射漆树酸5 mg/kg;ALI+T组和ALI+T+A组注射LPS前30 min时腹腔注射TZD 50 mg/kg。给予LPS 12 h后处死小鼠取肺组织, 测定湿重/干重(W/D)比值, 光镜下观察病理学结果, 并行肺损伤评分;分别采用Western blot法和PCR法测定PIAS1、PIAS2、PIAS3和PIASy及其mRNA的表达。实验Ⅱ 体外培养的小鼠肺泡巨噬细胞(MH-S细胞)采用随机数字表法分为4组(n=5):对照组(C组)、LPS...     

右美托咪定通过抑制JAK2/STAT3通路改善脂多糖诱导的小鼠急性肺损伤

目的研究右美托咪定对脂多糖(LPS)诱导的急性肺损伤(ALI)小鼠JAK2/STAT3通路的作用。方法雄性昆明小鼠36只,随机均分为三组:正常组(C组)、模型组(L组)和右美托咪定预处理组(D组)。取小鼠右肺下叶,称量并计算湿/干比重(W/D),对右肺上叶进行苏木素-伊红(HE)染色观察小鼠肺组织病理学改变,BCA法检测支气管肺泡灌洗液(BALF)中蛋白含量,用ELISA试剂盒检测BALF上清中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)及髓过氧化物酶(MPO)水平;qRT-PCR定量分析肺组织中TNF-α、IL-6mRNA水平;Western blot方法检测肺组织中p-JAK2、JAK2、p-STAT3、STAT3蛋白的含量。结果与L组比较,C组、D组肺损伤评分明显降低、小鼠肺W/D、BALF中蛋白含量明显减少,BALF中TNF-α、IL-6及MPO浓度明显降低,肺组织TNF-α和IL-6mRNA表达明显减少,p-JAK2/JAK2和p-STAT3/STAT3相对量明显减少(P0.05)。结论右美托咪定可能通过抑制JAK2/STAT3通路减轻LPS诱导的小鼠ALI。     

下丘脑外侧区星形胶质细胞NLRP3在小鼠失血性休克复苏后焦虑样行为中的作用

目的评价下丘脑外侧区星形胶质细胞NOD样受体相关蛋白3(NLRP3)在小鼠失血性休克复苏后焦虑样行为中的作用。方法清洁级雄性C57BL/6小鼠48只, 10周龄, 体质量25～30 g, 采用随机数字表法分为4组(n=12):假手术组(C组)、失血性休克复苏组(H组)、失血性休克复苏+腺病毒组(HI组)和失血性休克复苏+对照腺病毒组(HIV组)。H组、HI组和HIV组小鼠通过股静脉放血-回输法建立失血性休克复苏模型;建模前21 d时, HI组小鼠在立体定位仪引导下向双侧下丘脑外侧区注射AAV-GfaABC1D-EGFP-Cre腺病毒, HIV组注射AAV-GfaABC1D-EGFP对照腺病毒。复苏后14 d时, 采用埋珠实验和高架十字迷宫实验评估焦虑样行为;行为学实验结束后立即处死小鼠, 取含有下丘脑外侧区的脑组织, 采用免疫荧光染色法, 测定紫藤凝集素荧光强度反映细胞外基质表达, 测定NLRP3和胶质纤维酸性蛋白(GFAP)共定位情况, 计算cleaved caspase-1/GFAP、IL-18/GFAP阳性细胞数占总细胞的百分比。结果与C组比较, H组、HI组和HIV组埋珠数量减...     

Endotoxin tolerance from lipopolysaccharide pretreatment induces nuclear factor-kappaB alterations not present in C3H/HeJ mice

BACKGROUND: Lipopolysaccharide (LPS) activation of macrophage (MO) cytokine secretion requires activation and translocation of nuclear factor-kappaB (NF-kappaB). Endotoxin tolerance induced in LPS-responsive C3H/HeN MOs by LPS pretreatment results in decreased tumor necrosis factor (TNF) secretion and altered NF-kappaB activation. C3H/HeJ MOs have a genetic defect that renders them tolerant to LPS activation. We hypothesized that the alterations of NF-kappaB activation seen with LPS tolerance in HeN MOs would be present in HeJ mice. METHODS: MOs from C3H/HeJ and C3H/HeN mice were cultured with +/- 10 ng/mL LPS pretreatment for 24 hours and then stimulated with 1 to 1,000 ng/mL LPS. Activation of NF-kappaB was assayed by gel shift using a 32P-labeled specific oligonucleotide 30 minutes after LPS activation. TNF secretion 6 hours after LPS stimulation was measured by bioassay. RESULTS: LPS stimulation activated NF-kappaB in both HeN and HeJ MOs. We observed decreased NF-kappaB activation and a characteristic mobility shift in endotoxin-tolerant MOs from HeN mice that were not present in HeJ MOs. In contrast with the results in HeN mice, LPS pretreatment did not induce any alterations in NF-kappaB activation in HeJ MOs. LPS-stimulated TNF secretion was decreased in HeN MOs after LPS pretreatment. There was no change in TNF secretion in HeJ MOs, but, overall, TNF secretion by these cells was much less than that seen in HeN cells. CONCLUSION: MOs from C3H/HeN mice rendered LPS-tolerant by low-dose LPS pretreatment have alterations in activation of NF-kappaB not present in LPS-hyporesponsive C3H/HeJ mice.     

戊乙奎醚预先给药对大鼠急性肺损伤时NF-κB的影响

目的 探讨戊乙奎醚预先给药对大鼠急性肺损伤(ALI)时NF-κB的影响.方法 雄性SD大鼠35只,体重210～280 g,随机分为5组(n=7):对照组(C组)、ALI组和低、中、高剂量戊乙奎醚组(P1-3组).采用经尾静脉注射内毒素5 mg/kg的方法建立大鼠ALI模型.C组和ALI组经腹腔注射生理盐水0.5 ml,P1～3组分别经腹腔注射戊乙奎醚0.03、0.1和3 mg/kg,30 min后ALI组、P1～3组制备AU模型,C组不制备模型.于静脉注射LPS后4 h时,行血气分析,计算氧合指数;称量肺组织湿重(W)、干重(D),计算W/D;采用比色法测定肺组织髓过氧化物酶(MPO)活性;采用RT-PCR法测定肺组织肿瘤坏死因子-α(TNF-α)mRNA、白细胞介素-1β(IL-1β)mRNA的表达水平;采用ELISA法测定肺组织TNF-α和IL-1β的含量;采用非放射性EMSA法测定肺组织NF-κB活性;采用免疫组织化学法测定肺组织NF-κB的表达水平.结果 与C组比较,其余各组氧合指数降低,肺组织W/D和MPO活性、TNF-α,IL-1β含量及其相应mRNA表达水平、NF-κB活性和表达水平均升高(P<0.05或0.01);与ALI组比较,P1～3组氧合指数升高,肺组织W/D和MPO活性降低,TNF-α、IL-1β含量及其相应mRNA表达水平降低,NF-κB活性和表达水平降低(P<0.05);P1～3组上述指标比较差异无统计学意义(P>0.05).结论 戊乙奎醚预先给药减轻大鼠急性肺损伤的机制可能与抑制NF-κB的活化,下调NF-κB的表达,降低肺组织炎性反应有关.     

Neither Fas ligand nor endotoxin is responsible for inducible peritoneal phagocyte apoptosis during sepsis/peritonitis

Regulation of the phagocyte apoptotic response appears to play a significant role in the pathophysiology of sepsis. In this regard, prior studies have shown that the onset of phagocyte apoptosis, as well as those agents that regulate it at the nidus of infection, differ significantly from those seen in circulation. The aim of this study therefore was to determine if the increase in inducible phagocyte apoptosis and caspase activities seen in the peritoneum during sepsis is due to endotoxin or Fas ligand. To study this, male C3H/HeN (endotoxin-sensitive), C3H/HeJ (endotoxin-tolerant), and C3H/HeJ-FasL(gld) (endotoxin-tolerant/FasL-deficient) mice were subjected to cecal ligation and puncture or sham operation. Twenty-four hours later, phagocytes were collected and cultured with lipopolysaccharide (LPS), then harvested for apoptosis (propidium iodide cell cycle or cell death ELISA analysis), cytokine release (ELISA), and caspase activity (fluorogenic assay) determination. The data indicate that there was a marked increase in apoptosis in LPS-stimulated phagocytes which was associated with a significant increase in caspase 3, 8, and 9 activities but a decrease in caspase 1 activity from C3H/HeN and C3H/HeJ-FasL(gld) septic mice and an increase in caspase 3 and 8 activities in phagocytes from C3H/HeJ septic mice. Furthermore, cells from septic mice, including all three strains, lost their ability to produce IL-1beta and IL-6 in response to LPS stimulation. The inability to completely suppress these changes suggests that neither endotoxin (via signaling through TLR-4 pathway) nor Fas ligand regulates the peritoneal phagocyte apoptotic responses seen during the late phase of polymicrobial sepsis/peritonitis.     

促红细胞生成素预先给药对大鼠内毒素性急性肺损伤的影响

目的 探讨促红细胞生成素(EPO)预先给药对大鼠内毒素性急性肺损伤的影响.方法 成年雄性SD大鼠32只,体重180～220 g,随机分为4组(n=8),C组腹腔注射生理盐水4 ml/kg(EPO溶剂对照),30 min后静脉注射生理盐水2 ml/kg[脂多糖(LP3)溶剂对照];EPO组腹腔注射EPO3 000 U/kg,30 min后静脉注射生理盐水2 ml/kg;LPS组腹腔注射生理盐水4 ml/kg,30 min后静脉注射LPS 6 mg/kg;EPO+LPS组腹腔注射EPO 3 000 U/kg,30 min后静脉注射LPS 6 mg/kg.于静脉注射LPS后4 h时处死大鼠,观察肺组织病理学结果 ,计算肺组织湿/干重(W/D)比;测定肺组织髓过氧化物酶(MPO)活性和丙二醛(MDA)、一氧化氮(NO)含量;采用Western blot法测定肺组织诱导型一氧化氮合酶(iNOS)和硝基酪氨酸(NT)的表达.结果 与C组相比,LPS组和EPO+LPs组肺组织W/D比、MPO活性、MDA和NO含量升高,iNOS和NT表达上调(P<0.01);与LPS组相比,EPO+LPS组肺组织W/D比、MPO活性、MDA和NO含量降低,iNOS和NT表达下调(P<0.01).结论 EPO预先给药可减轻大鼠内毒素性急性肺损伤,与其下调iNOS表达,减少NO生成有关.     

阿米洛利预先给药对大鼠内毒素性急性肺损伤的影响

目的 探讨阿米洛利预先给药对大鼠内毒素性急性肺损伤的影响.方法 清洁级雄性SD大鼠32只,体重200～250 g,随机分为4组(n=8):对照组(C组)、急性肺损伤组(ALI组)、阿米洛利组(A组)和阿米洛利预先给药组(AL组).C组股静脉输注生理盐水3 ml,ALI组股静脉输注生理盐水1 ml、内毒素6 mg/kg,A组股静脉输注阿米洛利10 mg/kg、生理盐水2 ml,AL组股静脉输注阿米洛利10 mg/kg、内毒素6 mg/kg,输注速率均为0.05 ml/rain,给药间隔均为30 min.于输注内毒素结束后6 h时处死大鼠取肺,观察肺组织病理学,并行病理学评分,称重后计算肺湿干重比,检测髓过氧化物酶(MPO)活性,测定支气管肺泡灌洗液总蛋白、TNF-α和巨噬细胞炎性蛋白-2(MIP-2)的浓度,采用Western blot法检测肺组织钠氢交换体1(NHE1)、p38丝裂原活化蛋白激酶(p38MAPK)和细胞外信号调节激酶(ERK)的表达水平.结果 与C组比较,ALI组和AL组肺组织病理学评分、肺湿干重比、MPO活性、支气管肺泡灌洗液总蛋白、TNF-α和MIP-2浓度、肺组织NHE1、p38MAPK和ERK的表达水平明显升高(P＜0.01),A组上述指标差异无统计学意义(P＞0.05);与ALI组比较,AL组肺组织病理学评分、肺湿干重比、MPO活性、支气管肺泡灌洗液总蛋白、TNF-α和MIP-2浓度、肺组织NHE1和ERK的表达水平明显降低(P＜0.01),p38MAPK表达差异无统计学意义(P＞0.05).结论 阿米洛利预先给药可减轻大鼠内毒素性急性肺损伤,其机制可能与抑制ERK信号转导通路激活有关.     

c-Jun氨基末端激酶在大鼠内毒素性急性肺损伤中的作用

目的 评价c-Jun氨基末端激酶(JNK)在大鼠内毒素性急性肺损伤中的作用.方法 雄性成年SD大鼠80只,体重250～300 g,采用随机数字表法,将其随机分为4组(n=20):对照组(C组)、急性肺损伤组(ALI组)、SP600125组(S组)和二甲基亚砜组(D组).ALI组、S组和D组尾静脉注射LPS 5 mg/kg,C组尾静脉注射等容量生理盐水;S组和D组给予LPS后,分别尾静脉注射JNK抑制剂SP600125 30 mg/kg或二甲基亚砜0.2 ml.于给予LPS后4 h时,各组处死10只大鼠,回收支气管肺泡灌洗液(BALF)并取肺组织,采用ELISA法检测BALF中TNF-α和IL-1β的浓度,计算肺组织湿重/干重比(W/D比),观察肺组织病理学结果,并进行肺损伤评分.各组其余10只大鼠观察至给予LPS后48 h,记录大鼠生存情况.结果 与C组比较,其余各组BALF中TNF-α和IL-1β的浓度、肺组织W/D比和肺损伤评分升高,生存率降低(P<0.05或0.01);与ALI组比较,S组BALF中TNF-α和IL-1度、肺组织W/D比和肺损伤评分降低,生存率升高(P<0.01),D组差异无统计学意义(P>0.05).结论 JNK的活化参与了大鼠内毒素性急性肺损伤的发生发展.

Abstract：

Objective To evaluate the role of c-Jun N-terminal kinase (JNK) in lipopolysaccharide (LPS)-induced acute lung injury ( ALI) in rats.Methods Eighty male SD rats weighing 250-300 g were randomly divided into 4 groups ( n = 20 each) : control group (group C) ; ALI group; LPS + SP600125 (JNK inhibitor)group (group S) and LPS+ DMSO (the solvent) group (group DMSO) . ALI was induced by intravenous LPS 5mg/kg. In S and DMSO groups, SP600125 30 mg/kg and DMSO 0.2 ml were injected intravenously after LPS administration respectively. Ten animals were sacrificed by exsanguinafions at 4 h after LPS administration in each group. The broncho-alveolar lavage fluid (BALF) was colleted. The TNF-α and IL-1β concentrations in BALF were measured. The lungs were removed for microscopic examination and determination of W/D lung weight ratio. The other 10 animals in each group were observed for 48 h survival rate. Results Intravenous LPS significantly increased TNF-α and IL-1β concentrations in BALF and W/D lung weight ratio, decreased 48 h survival rate and induced histologic damage. Intravenous SP600125 30 mg/kg significantly attenuated the above-mentioned LPS-induced changes. Conclusion Activation of JNK is involved in the development of endotoxin-induced ALI in rats.