盐酸戊乙奎醚对内毒素性急性肺损伤大鼠肺组织Toll样受体4mRNA和Toll样受体2mRNA表达的影响

Objective To investigate the effect of penehyclidine (PHCD) on Toll-like receptor 4 (TLR4)mRNA and Toll-like receptor 2 (TLR2) mRNA expression in the lung tissue in rats with acute lung injury induced by lipopolysaccharide (LPS) .Methods Sixty healthy SD rats of both sexes weighing 200-220 g were randomly divided into 5 groups ( n = 12 each) :control group (group C) , LPS group and P1-3 groups. Acute lung injury was induced by intraperitoneal (IP) LPS 8 mg/kg in LPS and P1-3 groups. PHCD 0.3, 1.0 and 3.0 mg/kg were given IP after LPS administration in P1-3 groups. The animals were anesthetized at 6 h after IP LPS. Blood samples were collected for determination of serum TNF-α and IL-6 concentrations ( by ELISA) and then sacrificed, the lungs were immediately removed for determination of TLR4 mRNA and TLR2 mRNA expression (by RT-PCR), and microscopic examination. Results LPS significantly increased TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and IL-6 concentrations. PHCD 1.0 or 3.0 mg/kg significantly inhibited LPS-induced increase in TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and ILr6 concentrations.The lung histopathologic damage was significantly ameliorated in P2 and P3 groups as compared with group LPS.Conclusion PHCD can protect the lungs against LPS-induced acute lung injury through inhibiting TLR4 mRNA and TLR2 mRNA expression in the lung tissue and reducing the inflammatory response.     

盐酸戊乙奎醚对内毒素性急性肺损伤大鼠肺组织Toll样受体4mRNA和Toll样受体2mRNA表达的影响

Objective To investigate the effect of penehyclidine (PHCD) on Toll-like receptor 4 (TLR4)mRNA and Toll-like receptor 2 (TLR2) mRNA expression in the lung tissue in rats with acute lung injury induced by lipopolysaccharide (LPS) .Methods Sixty healthy SD rats of both sexes weighing 200-220 g were randomly divided into 5 groups ( n = 12 each) :control group (group C) , LPS group and P1-3 groups. Acute lung injury was induced by intraperitoneal (IP) LPS 8 mg/kg in LPS and P1-3 groups. PHCD 0.3, 1.0 and 3.0 mg/kg were given IP after LPS administration in P1-3 groups. The animals were anesthetized at 6 h after IP LPS. Blood samples were collected for determination of serum TNF-α and IL-6 concentrations ( by ELISA) and then sacrificed, the lungs were immediately removed for determination of TLR4 mRNA and TLR2 mRNA expression (by RT-PCR), and microscopic examination. Results LPS significantly increased TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and IL-6 concentrations. PHCD 1.0 or 3.0 mg/kg significantly inhibited LPS-induced increase in TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and ILr6 concentrations.The lung histopathologic damage was significantly ameliorated in P2 and P3 groups as compared with group LPS.Conclusion PHCD can protect the lungs against LPS-induced acute lung injury through inhibiting TLR4 mRNA and TLR2 mRNA expression in the lung tissue and reducing the inflammatory response.     

盐酸戊乙奎醚对内毒素性急性肺损伤大鼠肺组织Toll样受体4mRNA和Toll样受体2mRNA表达的影响

目的 探讨盐酸戊乙奎醚对内毒索性急性肺损伤大鼠肺组织Toll样受体4(TLR4)mRNA和Toll样受体2(TLR2)mRNA表达的影响.方法 健康SD大鼠60只,雌雄不拘,体重200～220g,采用随机数字表法,将大鼠随机分为5组(n=12),对照组(C组)、LPS组和低、中、高剂量盐酸戊乙奎醚组(P1组～P3组).C组腹腔注射生理盐水2ml;LPS组腹腔注射LPS 8mg/kg;P1组～P3组分别腹腔注射LPS 8 mg/kg和盐酸戊乙奎醚0.3、1.0和3.0 mg/kg.给药结束后6 h时开胸,心室取血,并取肺组织,采用ELISA法测定血清TNF-α和Ib-6的浓度,RT-PCR法测定肺组织TLR4 mRNA和TLR2 mRNA 的表达水平,并观察肺组织病理学结果.结果 与C组比较,LPS组、P1组～P3组血清TNF-α、IL-6浓度和肺组织TLR4 mRNA、TLR2 mRNA表达均升高(P<0.05);与LPS组比较,P2组和P3组血清TNF-α、IL-6浓度和肺组织TLR4 mRNA、TLR2 mRNA表达均降低(P<0.05),P1组上述指标差异无统计学意义(P>0.05);与P1组比较,P2组和P1组血清TNF-α、IL-6浓度和肺组织TLR4 mRNA、TLR2 mRNA表达均降低(P<0.05);P2组和P3组血清TNF-α、IL-6浓度和肺组织TLR4 mRNA、TLR2 mRNA表达比较差异无统计学意义(P>0.05).P2组和P3组肺组织病理学损伤程度明显轻于LPS组.结论 盐酸戊乙奎醚可通过下调肺组织TLR4 mRNA和耵JR2 mRNA的表达,降低炎性反应,从而减轻大鼠内毒素性急性肺损伤.

Abstract：

Objective To investigate the effect of penehyclidine (PHCD) on Toll-like receptor 4 (TLR4)mRNA and Toll-like receptor 2 (TLR2) mRNA expression in the lung tissue in rats with acute lung injury induced by lipopolysaccharide (LPS) .Methods Sixty healthy SD rats of both sexes weighing 200-220 g were randomly divided into 5 groups ( n = 12 each) :control group (group C) , LPS group and P1-3 groups. Acute lung injury was induced by intraperitoneal (IP) LPS 8 mg/kg in LPS and P1-3 groups. PHCD 0.3, 1.0 and 3.0 mg/kg were given IP after LPS administration in P1-3 groups. The animals were anesthetized at 6 h after IP LPS. Blood samples were collected for determination of serum TNF-α and IL-6 concentrations ( by ELISA) and then sacrificed, the lungs were immediately removed for determination of TLR4 mRNA and TLR2 mRNA expression (by RT-PCR), and microscopic examination. Results LPS significantly increased TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and IL-6 concentrations. PHCD 1.0 or 3.0 mg/kg significantly inhibited LPS-induced increase in TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and ILr6 concentrations.The lung histopathologic damage was significantly ameliorated in P2 and P3 groups as compared with group LPS.Conclusion PHCD can protect the lungs against LPS-induced acute lung injury through inhibiting TLR4 mRNA and TLR2 mRNA expression in the lung tissue and reducing the inflammatory response.     

盐酸戊乙奎醚预先给药对新生大鼠内毒素性急性肺损伤时NF-κB活性的影响

Objective To investigate the effects of penehyclidine (PHCD) pretreatment on nuclear factor kappa B ( NF-kB ) activity during lipopolysaccharide ( LPS )-induced acute lung injury ( ALl ) in neonate rats.Methods Thirty 7-day old Wistar rats of both sexes weighing 18-21 g were randomly divided into 3 groups ( n =10 each): group Ⅰ control (group C); group Ⅱ LPS; group Ⅲ PHCD. Group Ⅱ and Ⅲ received intraperitoneal ( group IP) LPS 3 mg/kg. In group Ⅲ PHCD 5 mg/kg was administered IP at 30 min before LPS respectively. The animals were killed at 4 h after LPS administration. The lungs were immediately removed. The W/D lung weight ratio was measured. The TNF-α, IL-1 βand IL-10 content in the lung were detected by ELISA and expression of NF-kB p65 was detected by immuno-histochemical staining.Results LPS significantly increased W/D lung weight ratio, TNF-α, IL-1 β, IL-10 content and NF-kB p65 expression in the lung as compared with control group. PHCD administered before LPS significantly attenuated the LPS-induced changes. Electron microscopy showed that PHCD before LPS significandy ameliorated the LPS-induced histological damages. Conclusion Pretreatment with PHCD can attenuate LPS-induced acute lung injury though inhibition of NF-kB activation and inflammatory response of lung tissue in neonate rats.     

盐酸戊乙奎醚预先给药对新生大鼠内毒素性急性肺损伤时NF-κB活性的影响

Objective To investigate the effects of penehyclidine (PHCD) pretreatment on nuclear factor kappa B ( NF-kB ) activity during lipopolysaccharide ( LPS )-induced acute lung injury ( ALl ) in neonate rats.Methods Thirty 7-day old Wistar rats of both sexes weighing 18-21 g were randomly divided into 3 groups ( n =10 each): group Ⅰ control (group C); group Ⅱ LPS; group Ⅲ PHCD. Group Ⅱ and Ⅲ received intraperitoneal ( group IP) LPS 3 mg/kg. In group Ⅲ PHCD 5 mg/kg was administered IP at 30 min before LPS respectively. The animals were killed at 4 h after LPS administration. The lungs were immediately removed. The W/D lung weight ratio was measured. The TNF-α, IL-1 βand IL-10 content in the lung were detected by ELISA and expression of NF-kB p65 was detected by immuno-histochemical staining.Results LPS significantly increased W/D lung weight ratio, TNF-α, IL-1 β, IL-10 content and NF-kB p65 expression in the lung as compared with control group. PHCD administered before LPS significantly attenuated the LPS-induced changes. Electron microscopy showed that PHCD before LPS significandy ameliorated the LPS-induced histological damages. Conclusion Pretreatment with PHCD can attenuate LPS-induced acute lung injury though inhibition of NF-kB activation and inflammatory response of lung tissue in neonate rats.     

c-Jun氨基末端激酶在大鼠内毒素性急性肺损伤中的作用

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.     

c-Jun氨基末端激酶在大鼠内毒素性急性肺损伤中的作用

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.     

骨髓间充质干细胞对急性出血坏死性胰腺炎大鼠肺组织Toll样受体2/4mRNA表达的影响及意义

目的 研究骨髓间充质干细胞(BMSCs)对急性出血坏死性胰腺炎(AHNP)大鼠肺组织T0ll样受体(TLR)2/4表达的影响并初步探讨其机制.方法 采用逆行胰胆管牛磺胆酸钠注射制造AHNP大鼠模型,动物分为假手术组、胰腺炎组和BMSCs治疗组;流式细胞仪检测BMSCs表面标记阳性细胞率;RT-PCR方法检测肺组织TLR2/4mRNA表达变化;同时观察肺组织形态学改变,进行肺湿/干重比(W/D)测定.结果 与假手术组比较,胰腺炎组大鼠从3h时肺组织TLR2/4mRNA表达开始增高,在12 h时肺组织TLR2/4mRNA表达达到峰值;同时肺损伤加重,肺组织TNF-α浓度升高(P＜0.05).给予BMSCs治疗后,TLR2/4mRNA表达降低,肺损伤程度减轻,肺组织TNF-α浓度降低(P＜0.05).结论 急性出血坏死性胰腺炎时,组织内TLR2和TLR4mRNA表达上调,肺组织损伤加重.BMSCs可以明显抑制AHNP肺组织TLR2/4mRNA的表达,降低肺组织TNF-α浓度,从而减轻肺损伤.

Abstract：

Objective To investigate the effect of bone mesenchymal stem cells on Toll-like receptors (TLR) 2/4 expression in the lungs of rats with acute hemorrhagic necrotizing pancreatitis (AHNP). Methods Seventy SD male rats were randomly divided into sham-operation group (n=10), AHNP group(n=30) and MSCs-treated group(n=30). Masc rate of BMSCs with surface mark were measured by flow cytometer. TLR2/4mRNA expression in the the lung were measured by RT-PCR, and The ratio of Wet/dry and lung histological changs were observed. Results TLR2/4 mRNA could be detected in the lungs with low values in sham-operation group, markedly increased in 3 h, and peaked in 12 h in AHNP group (P＜0.05). Lung injuries were aggravated and the levels of TNF-α in the lung were increased (P＜0. 05) . Treatment with MSCs could effectively inhibit TLR2/4 mRNA expression and relieve lung injuries. The levels of TNF-α in the lung were decreased (P＜0.05). Conclusions The expression of TLR2/4 mRNA is increased in the lungs in AHNP and the lung injuries are aggravated. MSCs could markedly inhibit TLR2/4 mRNA expression in the lungs in AHNP, which would lead to relief of lung injury.     

吸入一氧化碳对大鼠脑死亡致肺损伤的影响

Objective To investigate the effects of carbon monoxide (CO) inhalation on lung injury induced by brain death (BD) in rats. Methods Adult male Wistar rats weighing 250-300 g were used in this study. The animals were anesthetized with intraperitoneal pentobarbital sodium 60 mg/kg, tracheostomized and mechanically ventilated (VT 10 ml/kg, RR 50 bpm, PEEP 2 cm H2O). A balloon-tip catheter was placed in the cranium. Twenty-four rats in which Fogarty catheter was successfully placed in the cranium without complication were randomly divided into 3 groups ( n = 8 each) : group I sham operation (group S) ; group II BD and group Ⅲ BDCO. BD was induced by increase in intracranial pressure produced by inflating the balloon at the tip of the catheter. In group S the balloon of the catheter was not inflated. The animals inhaled 40% O2 for 150 min. In group BD, BD was induced and confirmed at 30 min after inflation of the balloon. Then 40% O2 was inhaled for 120 min. In group BDCO, 40% O2 and 0.025% CO were inhaled for 120 min after BD was confirmed at 30 min after balloon inflation. At the end of the experiment the animals were killed. Arterial blood samples were obtained for blood gas analysis before anesthesia (basline), immediately after confirmation of BD, and at 30, 60, 90 and 120 min of CO inhalation. Blood was collected for determination of plasma TNF-α, IL-6 and IL-10 concentrations at 120 min of CO inhalation. The lungs were obtained for determination of W/D lung weight ratio, and MPO activity in the lung tissue and microscopic examination. Lung injury scores were calculated. Results PaO2/FiO2 was stable during the 150 min in group S. Brain death significantly decreased PaO2/FiO2 at 30 min after balloon inflation. PaO2/FiO2 was gradually decreasing during the 120 min in group BD. CO inhalation prevented PaO2/FiO2 from decreasing further. W/D lung weight ratio and MPO activity were significantly higher in group BD than in group S and BDCO. The lung injury score (1 = normal, 4= severely injured) and plasma TNF-αα IL-6 and IL-10 concentrations were significantly higher in group BD than in group S. CO inhalation ameliorated the BD-induced lung injury and attenuated the increase in plasma TNF-a and IL-6 concentration. Plasma IL-10 concentration was significantly higher in group BDCO than in group BD. Conclusion CO inhalation can ameliorate acute lung injury induced by BD through decreasing the local and systemic inflammatory response.     

七氟烷对急性肺损伤大鼠肺组织细胞凋亡的影响及其机制

Objective To investigate the effect and the possible mechanism of sevoflurane on lipopolysaccharide (LPS)induced acute lung injury (ALI) in rats. Methods eighteen male SD rats were randomly divided into three groups. The ALI group received LPS 5 mg/kg, while the control group received normal saline,and the sevoflurane and LPS group received sevoflurane (2.5%)for 30min after ALI induced by LPS 5 mg/kg. Lung tissue samples were taken at 12 h after instillation of LPS, histological examination by lung water contant (wet/dry ,W/D) calculation and light microcopy was performed. Apoptosis was determined by flow cytometry (FCM),TUNEL test (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) and electron microscope. At the same time, Fas, Bcl-2 protein expression were studied by using immunocytochemistry and western blot techniques in all groups mentioned above. Results The change of the levels of W/D(6.74±0.26), AI(48.1±1.9,46.6±1.2), Fas(0.223±0.034), Bcl-2(28.5±0.5) of the LPS group was significant compared with the control group (11.2±2.3,8.6±0.5,4.35±0.37,0.091±0.013,75.6±2.7), and lung injury was more severe. The levels of W/D(5.37±0.23), AI(27.2±0.9,26.3±2.4), Fas of the sevofluran group(0.131±0.21 ) were decreased, while,Bcl-2 expression (81.2±5.2) were increased compared with the LPS group (0.223±0.034,28.5±0.5)(P＜0.01), and lung injury was attenuated. Conclusion Sevoflurane inhalation protects lung from injury by inhibiting excessive cell apoptosis, Fas expression and up-regulating Bcl-2 expression, which may play an important role in the pathogenesis of LPS-induced ALI.     

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.     

盐酸戊乙奎醚预先给药对失血性休克大鼠急性肺损伤时TLR4 mRNA表达的影响

目的 探讨盐酸戊乙奎醚预先给药对失血性休克大鼠急性肺损伤时Toll样受体4(TLR4)mRNA表达的影响.方法 健康SD大鼠40只,体重200～250 g,随机分为5组(n=8):假手术组(S组)、失血性休克致急性肺损伤组(ALI组)和低、中、高剂量盐酸戊乙奎醚预先给药组(P1～3组).S组仅行动静脉穿刺,不放血,ALI组股动脉放血至35～45 mm Hg制备急性肺损伤模型,P1～3组分别于放血前30 min股静脉注射盐酸戊乙奎醚0.3、1.0、3.0 mg/kg,随后制备急性肺损伤模型.各组复苏后4 h时处死大鼠取肺,称重后计算肺湿干重比,检测TLR4 mRNA和NF-κB p65蛋白的表达水平,观察病理学结果.结果 与S组比较,ALI组和P1组TLR4 mRNA、NF-κB p65蛋白表达水平及肺湿干重比升高(P＜0.05或0.01),P2.3组差异无统计学意义(P＞0.05);与ALI组比较,P2,3组TLR4 mRNA、NF-κB p65蛋白表达水平及肺湿干重比降低(P＜0.05或0.01);P2组和P3组上述指标比较差异无统计学意义(P＞0.05).P2,3组肺组织病理学损伤程度较ALI组明显减轻.结论 盐酸戊乙奎醚预先给药可通过抑制肺组织TLR4 mRNA表达上调,进而降低NF-κB活性,从而减轻失血性休克诱发大鼠的急性肺损伤.     

盐酸戊乙奎醚预先给药对内毒素性急性肺损伤大鼠肺组织NF-κB mRNA表达及SOD活性的影响

目的 探讨盐酸戊乙奎醚预先给药对内毒素性急性肺损伤大鼠肺组织NF-κB mRNA表达及SOD活性的影响.方法 健康雄性SD大鼠32只,月龄2月,体重230～280 g,随机分为4组(n=8),对照组(C组)腹腔和尾静脉均注射生理盐水1 ml/kg;急性肺损伤组(ALI组):腹腔注射生理盐水1 ml/kg,30 min后经尾静脉注射LPS 5 mg/kg;盐酸戊乙奎醚低剂量组(LP组)、高剂量组(HP组)分别腹腔注射盐酸戊乙奎醚0.3和1 mg/kg,30 min后经尾静脉注射LPS 5 mg/kg.静脉注射生理盐水或LPS后6 h时,取肺组织,检测NF-κB mRNA的表达、TNF-α和MDA的含量和SOD活性,计算肺组织湿/干重比(W/D)及含水量,观察肺组织病理学结果.结果 与C组比较,ALI组、LP组和HP组肺组织NF-κB mRNA表达上调,TNF-α及MDA含量升高,SOD活性降低,W/D和肺组织含水量升高(P＜0.05);与ALI组比较,LP组和HP组肺组织NF-κB mRNA表达下调,TNF-α及MDA含量降低,SOD活性升高,W/D和肺组织含水量降低(P＜0.05);与LP组比较,HP组肺组织NF-κB mRNA表达下调,TNF-α及MDA含量降低,SOD活性升高,W/D和肺组织含水量降低(P＜0.05).LP组和HP组肺组织病理学损伤较ALI组减轻.结论 盐酸戊乙奎醚预先给药减轻大鼠内毒素性急性肺损伤的机制可能与下调肺组织NF-κB mRNA表达,降低肺局部炎性反应,增强机体抗氧化能力有关.     

盐酸戊乙奎醚预先给药对失血性休克大鼠急性肺损伤时NF-κB活性的影响

目的 探讨盐酸戊乙奎醚预先给药对失血性休克大鼠急性肺损伤时NF-κB活性的影响.方法 健康成年Wistar大鼠24只,体重200～250 g,雌雄不限,随机分为3组(n=8):假手术组(S组)、失血性休克致急性肺损伤组(ALI组)和盐酸戊乙奎醚预先给药组(P组).S组仅行动、静脉穿刺,不制备急性肺损伤模型;ALI组和P组经右侧颈内动脉穿刺置管监测BP,左侧股动脉置管放血,通过放血和回输血液维持BP 35～45 mm Hg 1 h,然后回输全部失血及等同于失血量的生理盐水,制备急性肺损伤模型;P组于放血前即刻静脉注射盐酸戊乙奎醚2 mg/kg. 于模型制备成功后6 h,采集右侧股动脉血样行血气分析,采用ELISA法测定右心房血浆TNF-α浓度,计算肺湿干重比,采用免疫组织化学法检测右肺组织NF-κB p65的表达,光镜下观察肺组织病理学.结果 与S组比较,Au组和P组PaO2降低,PaCO2、肺湿干重比、TNF-α浓度升高,NF-κB p65表达上调(P<0.05);与ALI组相比,P组PaO2升高,PaCO2、肺湿干重比、TNF-α浓度降低,NF-κB p65表达下调(P<0.05).病理结果显示:P组肺组织损伤较ALI组明显减轻.结论 盐酸戊乙奎醚预先给药可通过降低肺组织NF-κB的活性抑制炎性反应,从而减轻失血性休克诱发大鼠的急性肺损伤.     

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.     

雷公藤甲素对内毒素致大鼠急性肺损伤的影响

目的 评价雷公藤甲素对内毒素(LPS)致大鼠急性肺损伤的影响.方法 雄性SD大鼠65只,体重200 ～ 250 g,采用随机数字表法,将其随机分为5组,对照组(C组,n=5):尾静脉注射生理盐水,同时腹腔注射1％二甲基亚砜(DMSO);LPS组(L组,n=15)和不同剂量雷公藤甲素组(TP1～3组,n=15):尾静脉注射LPS 5 mg/kg,同时腹腔分别注射1％ DMSO和雷公藤甲素25、50、100μg/kg.于给药前1h和给药后1、3、6、12 h时行动脉血气分析;给药后12 h时心脏采血后处死大鼠,取肺组织,收集支气管肺泡灌洗液(BALF),采用ELISA法测定血清和BALF中TNF-α浓度;测定肺组织湿重(W)和干重(D),计算W/D比;光镜下观察肺组织病理学结果,并进行弥漫性肺泡损伤评分(DAD评分);采用荧光定量PCR法测定肺组织Toll样变体4(TLR4) mRNA表达;采用Western blot法测定肺组织TLR4蛋白表达.结果 与C组相比,L组和TP1～3组给药后3、6和12 h时PaO2下降,DAD评分及W/D比升高,L组、TP1组和TP2组血清和BALF中TNF-α浓度升高,肺组织TLR4 mRNA及其蛋白表达上调,TP3组血清和BALF中TNF-α浓度降低,肺组织TLR4mRNA及其蛋白表达下调(P＜0.05).与L组和TP1组相比,TP2组和TP3组给药后6和12 h时PaO2升高,DAD评分、W/D比、血清和BALF中TNF-α浓度降低,肺组织TLR4 mRNA及其蛋白表达下调(P＜0.05).与TP2组相比,TP3组血清和BALF中TNF-α浓度降低,肺组织TLR4 mRNA及其蛋白表达下调(P＜0.05).L组和TP1组间、TP2组和TP3组间血气指标、DAD评分及W/D比较差异无统计学意义(P＞0.05).TP1～3组肺组织病理学损伤较L组减轻.结论 雷公藤甲素可减轻LPS诱发的大鼠急性肺损伤,且与剂量有关,其机制与抑制TLR4表达的上调,减少TNF-α的释放有关.     

氟比洛芬酯预先给药对大鼠内毒素性急性肺损伤的影响

目的 评价氟比洛芬酯预先给药对大鼠内毒索性急性肺损伤的影响.方法 雄性SD大鼠40只,体重190～220 g,随机分为3组:对照组(C组,n=8)、脂多糖组(LPS组,n=16)和氟比洛芬酯组(FA组,n=16).C组尾静脉注射生理盐水(NS)1 ml;LPS组尾静脉注射LPS 5 mg/kg(1 m1);FA组尾静脉注射氟比洛芬酯6 mg/kg(1 m1)后0.5 h注射LPS 5 mg/kg(1 m1).LPS组和FA组于注射LPS后2、4 h时、C组于注射NS后4 h时,各随机处死8只大鼠,取股动脉血样8 ml,行血气分析;采用放免法测定血清血栓素B2(TXB2)和6-酮-前列腺素F1α(6-keto PGF1α)的浓度;采用ELISA方法测定血清肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-6和IL-10的浓度.取左肺上叶组织,计算肺组织湿/干重比值(W/D)和肺含水量(LC).注射LPS或NS后4 h时取左肺下叶组织,光镜下观察肺组织病理学结果.结果 与C组比较,LPS组PaO2和PaO2/FiO2下降,PaCO2、W/D和LC升高,血清TXB2、6-keto PGF1α浓度和TXB2/6-keto PGF1α比值升高,血清TNF-α、IL-1β和IL-6浓度升高,FA组W/D和LC升高,血清TXB2和6-keto PGF1α浓度升高,血清TNF-α、IL-1β、IL-6和IL-10浓度升高(P<0.05或0.01).与LPS组比较,FA组PaCO2、W/D和LC降低,PaO2和PaO2/FiO2升高,血清TXB2、6-keto PGF1α浓度和TXB2/6-keto PGF1α比值降低,血清IL-1β、IL-6和TNF-α浓度降低,血清IL-10浓度升高(P<0.05).FA组肺组织病理学损伤较LPS组轻.结论 氟比洛芬酯预先给药可减轻内毒素性急性肺损伤,可能与维持血液TXA2和PGI2平衡及抑制炎性反应有关.     

抑制低氧诱导因子-1α表达对小鼠肠缺血／再灌注所致肺损伤的影响

目的探讨通过低氧诱导因子-1α（hypoxiainduciblefactor-1α,HIF—1α）抑制剂YC-1预先抑制该基因表达对肠缺血,再灌注（ischemia／reperfusion,I／R）致急性肺损伤的影响。方法6周～8周龄健康雄性C57BL／6小鼠36只,采用随机数字表法随机分为3组（每组12只）：假手术组（S组）、I／R组和I／R±YC—I预处理组（YC—I组）。YC-1组于术前10min腹腔注入YC-1（1mg／kg）,采用夹闭C57BL／6小鼠肠系膜前动脉45min后再灌注6h的方法造成肠YR损伤模型,取小鼠肺标本称重后计算肺湿干重比,苏木素-伊红（hematoxylin-eosin,HE）染色后观察肺组织病理学改变,分光光度法测定髓过氧化物酶（myeloperoxidase,MPO）活性、酶联免疫吸附测定法检测肺组织肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）和白细胞介素（interleukin,IL）-1β表达,反转录-PCR法检测HIF-1α、Toll样受体4（toll—likereceptor4,TLR4）mRNA的表达。结果与I／R组比较,预先抑制HIFqct表达使肺实质水肿及中性粒细胞浸润聚集减少,肺组织病理学损伤减轻,肺湿干重比显著降低（RnOI）,MPO活性下调[（1．88±0．82）u／g],TNF-α[（187±20）ng／L）]、IL一1β[（536±54）ng／L）]、HIF-1α、TLR4mRNA的表达水平下降（P〈0．05）。结论YC-1预处理可使肺组织TLR4mRNA表达下调,抑制肠I／R肺组织中促炎细胞因子的释放,明显减轻小鼠肠I／R后急性肺损伤。     

蜕皮甾酮对急性肺损伤大鼠肺组织中Toll样受体4及肺表面活性蛋白A表达的影响

目的：观察蜕皮甾酮（EDS）对脂多糖（LPS）诱导急性肺损伤（ALI）大鼠肺组织中TNF-α、肺表面活性蛋白A（SP-A）、Toll样受体4（TLR4）表达的影响,并探讨其机制.方法：将40只雄性Wistar大鼠随机分成正常对照组、LPS组、EDS低（20 mg/kg）、中（30 mg/kg）、高（40 mg/kg）剂量治疗组（n=8）.腹腔注射LPS（10 mg/kg）诱导大鼠ALI模型,3个EDS治疗组于建模1 h后予不同剂量的EDS腹腔注射,其余两组注射等体积生理盐水.24 h后,取肺组织,用光学显微镜观察各组肺组织病理学改变;用Western blot测定肺表面活性蛋白A（SP-A）、Toll样受体4（TLR4）的表达;用ELISA测定各组肺组织中TNF-α含量,RT-PCR检测TNF-α mRNA表达水平.结果：肺组织病理学观察显示：LPS组可见肺间质充血、水肿,大量炎性细胞浸润,而EDS治疗组肺损伤明显改善,效果随EDS剂量的增加而增加.与正常对照组比较,LPS组肺组织的SP-A蛋白表达明显降低（P〈0.05）,而TNF-α含量、TNF-α mRNA表达、TLR4蛋白表达明显增加 （P〈0.05）.与LPS组相比较,不同剂量EDS治疗组肺组织SP-A蛋白表达均增加（P〈0.05）,而TNF-α含量、TNF-α mRNA表达、TLR4蛋白表达明显降低（P〈0.05）,其中EDS高剂量组比中、低剂量组效果明显 （P〈0.05）.结论：蜕皮甾酮对LPS诱导大鼠急性肺损伤有保护作用,其机制可能与抑制TLR4通路来降低肺组织中TNF-α炎性因子的表达,并促进抗炎物质SP-A释放有关.