吲哚胺2,3-双加氧酶在哮喘小鼠模型中的作用

目的 探讨吲哚胺2,3-双加氧酶(IDO)在哮喘小鼠模型中所起的作用.方法 卵白蛋白(OVA)致敏、激发诱导哮喘小鼠模型.分别检测肺组织中IDO在蛋白水平和mRNA水平上的表达.免疫荧光组织化学方法检测树突状细胞(DCs)在肺组织中的分布和成熟状态.结果 ①OVA诱导和激发的小鼠哮喘模型的症状和肺部炎症病理改变较对照组严重;哮喘组小鼠血清总IgE为(165.50 ± 30.13)ng/ml,对照组为(94.45 ± 28.30)ng/ml,差异有统计学意义(P ＜ 0.05).②哮喘组小鼠肺部IDO的表达低于对照组.以平均累积光密度为指标检测的小鼠肺组织中IDO在蛋白水平上的表达,哮喘组为11.38 ± 6.05,对照组为23.62 ± 8.92,差异有统计学意义(P ＜ 0.05);实时荧光定量PCR检测小鼠肺组织中IDO在mRNA水平上的表达,哮喘组是对照组的33%,显著低于对照组(P ＜ 0.05).③小鼠肺组织中CD11c+CD86+细胞主要分布于肺泡壁和小血管周围.以双标阳性细胞平均累积荧光强度为指标检测小鼠肺组织中CD11c+CD86+细胞数量,哮喘组的中位数为9 961.86(7 406.52 ～ 12 724.98),对照组为15 974.60(10 006.39 ～ 16 171.46),差异有统计学意义(P ＜ 0.05).结论 哮喘小鼠肺组织表达IDO低,且成熟树突状细胞减少.提示在哮喘小鼠中,由于成熟的树突状细胞较少,产生IDO偏少,这可能在哮喘发生中起着重要作用.

Abstract：

Objective To investigate the role of indoleamine 2,3-dioxygenase(IDO)in asthma.Methods The mouse asthma model was induced by ovalbumin(OVA).IDO expression was detected on the level of protein and mRNA respectively.Distribution and maturation of dendritic cells were detected by the immunofluorescence method.Results (1)The symptoms and lung inflammation in the model group were more serious than control group.The serum total IgE was significantly higher in the model group than that in the control group,165.50 ± 30.13 ng/ml vs.94.45 ± 28.30 ng/ml(P ＜ 0.05).(2)IDO expression in the model group was lower than that in control group.On the level of protein,mean intergrated optical density was 11.38 ± 6.05 in the model group vs.23.62 ± 8.92 in the control group(P ＜ 0.05);on the level of mRNA,IDO expression of the model group was 33% of the control group(P ＜ 0.05).(3)CD11c+CD86+ cells were distributed in alveolar wall and around small vessels.The quantity of CD11c+CD86+ cells in lungs of the model group were significantly smaller than that in the control group.The median intergrated fluorescence intensity was 9961.86(range,7 406.52 ～ 12 724.98)in the model group vs.15974.60(range,10 006.39 ～ 16 171.46)in the control group(P ＜ 0.05).Conclusions IDO expression is low and matured dendritic cells are less in situ in the asthma model.These suggest that less matured DCs may produce less IDO,which may play an important role in asthma.     

肺复张手法对急性肺损伤幼猪肺表面活性物质的影响

Objective To investigate the effect of recruitment maneuver(RM) on surfactant proteins in young piglets with acute lung injury and the possible mechanisms of lung recruitment after RM.Methods The piglet model of ALI was established by lipopolysaccharide intravenous injection,12 male piglets were randomly divided into two groups:conventional ventilation group(control group) and RM with low tidal volume group(RM group).After 8 hours of ventilation,mRNA expression of surfactant protein-A(SP-A),SP-B,SP-C,SP-D in the piglet lungs were determined by real time PCR and SP-A protein distribution was assessed by immunohistochemistry.Biochemical analyses of TP,total phospholipids(TPL),DSPC were conducted as well.SP-A levels in bronchoalveolar lavage fluid(BALF) and plasma were measured by enzymelinked immunosorbent assay(ELISA).Results As compared with control group,RM group had higher expression of SP-A,SP-B,SP-C and SP-D.SP-A average gray values of control group and RM group were 97.8±6.4 and 106.3±8.5,and there was significant difference(P＜0.01).RM group showed significant increase of TPL,DSPC and DSPC/TP.The concentration of SP-A in BALF was higher in RM group than that of the control group,however,SP-A plasma level was lower in RM group than that of the control group.Conclusion RM can increase suffactant protein expression in ALI animals,alleviate surfactant protein dysfunction and regulate the concentration of SP-A,which may improve alveolar recruitment following the RM and alleviate ventilator-induced lung injury.     

肺复张手法对急性肺损伤幼猪肺表面活性物质的影响

Objective To investigate the effect of recruitment maneuver(RM) on surfactant proteins in young piglets with acute lung injury and the possible mechanisms of lung recruitment after RM.Methods The piglet model of ALI was established by lipopolysaccharide intravenous injection,12 male piglets were randomly divided into two groups:conventional ventilation group(control group) and RM with low tidal volume group(RM group).After 8 hours of ventilation,mRNA expression of surfactant protein-A(SP-A),SP-B,SP-C,SP-D in the piglet lungs were determined by real time PCR and SP-A protein distribution was assessed by immunohistochemistry.Biochemical analyses of TP,total phospholipids(TPL),DSPC were conducted as well.SP-A levels in bronchoalveolar lavage fluid(BALF) and plasma were measured by enzymelinked immunosorbent assay(ELISA).Results As compared with control group,RM group had higher expression of SP-A,SP-B,SP-C and SP-D.SP-A average gray values of control group and RM group were 97.8±6.4 and 106.3±8.5,and there was significant difference(P＜0.01).RM group showed significant increase of TPL,DSPC and DSPC/TP.The concentration of SP-A in BALF was higher in RM group than that of the control group,however,SP-A plasma level was lower in RM group than that of the control group.Conclusion RM can increase suffactant protein expression in ALI animals,alleviate surfactant protein dysfunction and regulate the concentration of SP-A,which may improve alveolar recruitment following the RM and alleviate ventilator-induced lung injury.     

肺复张手法对急性肺损伤幼猪肺表面活性物质的影响

Objective To investigate the effect of recruitment maneuver(RM) on surfactant proteins in young piglets with acute lung injury and the possible mechanisms of lung recruitment after RM.Methods The piglet model of ALI was established by lipopolysaccharide intravenous injection,12 male piglets were randomly divided into two groups:conventional ventilation group(control group) and RM with low tidal volume group(RM group).After 8 hours of ventilation,mRNA expression of surfactant protein-A(SP-A),SP-B,SP-C,SP-D in the piglet lungs were determined by real time PCR and SP-A protein distribution was assessed by immunohistochemistry.Biochemical analyses of TP,total phospholipids(TPL),DSPC were conducted as well.SP-A levels in bronchoalveolar lavage fluid(BALF) and plasma were measured by enzymelinked immunosorbent assay(ELISA).Results As compared with control group,RM group had higher expression of SP-A,SP-B,SP-C and SP-D.SP-A average gray values of control group and RM group were 97.8±6.4 and 106.3±8.5,and there was significant difference(P＜0.01).RM group showed significant increase of TPL,DSPC and DSPC/TP.The concentration of SP-A in BALF was higher in RM group than that of the control group,however,SP-A plasma level was lower in RM group than that of the control group.Conclusion RM can increase suffactant protein expression in ALI animals,alleviate surfactant protein dysfunction and regulate the concentration of SP-A,which may improve alveolar recruitment following the RM and alleviate ventilator-induced lung injury.     

肺复张手法对急性肺损伤幼猪肺表面活性物质的影响

Objective To investigate the effect of recruitment maneuver(RM) on surfactant proteins in young piglets with acute lung injury and the possible mechanisms of lung recruitment after RM.Methods The piglet model of ALI was established by lipopolysaccharide intravenous injection,12 male piglets were randomly divided into two groups:conventional ventilation group(control group) and RM with low tidal volume group(RM group).After 8 hours of ventilation,mRNA expression of surfactant protein-A(SP-A),SP-B,SP-C,SP-D in the piglet lungs were determined by real time PCR and SP-A protein distribution was assessed by immunohistochemistry.Biochemical analyses of TP,total phospholipids(TPL),DSPC were conducted as well.SP-A levels in bronchoalveolar lavage fluid(BALF) and plasma were measured by enzymelinked immunosorbent assay(ELISA).Results As compared with control group,RM group had higher expression of SP-A,SP-B,SP-C and SP-D.SP-A average gray values of control group and RM group were 97.8±6.4 and 106.3±8.5,and there was significant difference(P＜0.01).RM group showed significant increase of TPL,DSPC and DSPC/TP.The concentration of SP-A in BALF was higher in RM group than that of the control group,however,SP-A plasma level was lower in RM group than that of the control group.Conclusion RM can increase suffactant protein expression in ALI animals,alleviate surfactant protein dysfunction and regulate the concentration of SP-A,which may improve alveolar recruitment following the RM and alleviate ventilator-induced lung injury.     

肺复张手法对急性肺损伤幼猪肺表面活性物质的影响

Objective To investigate the effect of recruitment maneuver(RM) on surfactant proteins in young piglets with acute lung injury and the possible mechanisms of lung recruitment after RM.Methods The piglet model of ALI was established by lipopolysaccharide intravenous injection,12 male piglets were randomly divided into two groups:conventional ventilation group(control group) and RM with low tidal volume group(RM group).After 8 hours of ventilation,mRNA expression of surfactant protein-A(SP-A),SP-B,SP-C,SP-D in the piglet lungs were determined by real time PCR and SP-A protein distribution was assessed by immunohistochemistry.Biochemical analyses of TP,total phospholipids(TPL),DSPC were conducted as well.SP-A levels in bronchoalveolar lavage fluid(BALF) and plasma were measured by enzymelinked immunosorbent assay(ELISA).Results As compared with control group,RM group had higher expression of SP-A,SP-B,SP-C and SP-D.SP-A average gray values of control group and RM group were 97.8±6.4 and 106.3±8.5,and there was significant difference(P＜0.01).RM group showed significant increase of TPL,DSPC and DSPC/TP.The concentration of SP-A in BALF was higher in RM group than that of the control group,however,SP-A plasma level was lower in RM group than that of the control group.Conclusion RM can increase suffactant protein expression in ALI animals,alleviate surfactant protein dysfunction and regulate the concentration of SP-A,which may improve alveolar recruitment following the RM and alleviate ventilator-induced lung injury.     

肺复张手法对急性肺损伤幼猪肺表面活性物质的影响

Objective To investigate the effect of recruitment maneuver(RM) on surfactant proteins in young piglets with acute lung injury and the possible mechanisms of lung recruitment after RM.Methods The piglet model of ALI was established by lipopolysaccharide intravenous injection,12 male piglets were randomly divided into two groups:conventional ventilation group(control group) and RM with low tidal volume group(RM group).After 8 hours of ventilation,mRNA expression of surfactant protein-A(SP-A),SP-B,SP-C,SP-D in the piglet lungs were determined by real time PCR and SP-A protein distribution was assessed by immunohistochemistry.Biochemical analyses of TP,total phospholipids(TPL),DSPC were conducted as well.SP-A levels in bronchoalveolar lavage fluid(BALF) and plasma were measured by enzymelinked immunosorbent assay(ELISA).Results As compared with control group,RM group had higher expression of SP-A,SP-B,SP-C and SP-D.SP-A average gray values of control group and RM group were 97.8±6.4 and 106.3±8.5,and there was significant difference(P＜0.01).RM group showed significant increase of TPL,DSPC and DSPC/TP.The concentration of SP-A in BALF was higher in RM group than that of the control group,however,SP-A plasma level was lower in RM group than that of the control group.Conclusion RM can increase suffactant protein expression in ALI animals,alleviate surfactant protein dysfunction and regulate the concentration of SP-A,which may improve alveolar recruitment following the RM and alleviate ventilator-induced lung injury.     

肺复张手法对急性肺损伤幼猪肺表面活性物质的影响

Objective To investigate the effect of recruitment maneuver(RM) on surfactant proteins in young piglets with acute lung injury and the possible mechanisms of lung recruitment after RM.Methods The piglet model of ALI was established by lipopolysaccharide intravenous injection,12 male piglets were randomly divided into two groups:conventional ventilation group(control group) and RM with low tidal volume group(RM group).After 8 hours of ventilation,mRNA expression of surfactant protein-A(SP-A),SP-B,SP-C,SP-D in the piglet lungs were determined by real time PCR and SP-A protein distribution was assessed by immunohistochemistry.Biochemical analyses of TP,total phospholipids(TPL),DSPC were conducted as well.SP-A levels in bronchoalveolar lavage fluid(BALF) and plasma were measured by enzymelinked immunosorbent assay(ELISA).Results As compared with control group,RM group had higher expression of SP-A,SP-B,SP-C and SP-D.SP-A average gray values of control group and RM group were 97.8±6.4 and 106.3±8.5,and there was significant difference(P＜0.01).RM group showed significant increase of TPL,DSPC and DSPC/TP.The concentration of SP-A in BALF was higher in RM group than that of the control group,however,SP-A plasma level was lower in RM group than that of the control group.Conclusion RM can increase suffactant protein expression in ALI animals,alleviate surfactant protein dysfunction and regulate the concentration of SP-A,which may improve alveolar recruitment following the RM and alleviate ventilator-induced lung injury.     

肺复张手法对急性肺损伤幼猪肺表面活性物质的影响

Objective To investigate the effect of recruitment maneuver(RM) on surfactant proteins in young piglets with acute lung injury and the possible mechanisms of lung recruitment after RM.Methods The piglet model of ALI was established by lipopolysaccharide intravenous injection,12 male piglets were randomly divided into two groups:conventional ventilation group(control group) and RM with low tidal volume group(RM group).After 8 hours of ventilation,mRNA expression of surfactant protein-A(SP-A),SP-B,SP-C,SP-D in the piglet lungs were determined by real time PCR and SP-A protein distribution was assessed by immunohistochemistry.Biochemical analyses of TP,total phospholipids(TPL),DSPC were conducted as well.SP-A levels in bronchoalveolar lavage fluid(BALF) and plasma were measured by enzymelinked immunosorbent assay(ELISA).Results As compared with control group,RM group had higher expression of SP-A,SP-B,SP-C and SP-D.SP-A average gray values of control group and RM group were 97.8±6.4 and 106.3±8.5,and there was significant difference(P＜0.01).RM group showed significant increase of TPL,DSPC and DSPC/TP.The concentration of SP-A in BALF was higher in RM group than that of the control group,however,SP-A plasma level was lower in RM group than that of the control group.Conclusion RM can increase suffactant protein expression in ALI animals,alleviate surfactant protein dysfunction and regulate the concentration of SP-A,which may improve alveolar recruitment following the RM and alleviate ventilator-induced lung injury.     

肺复张手法对急性肺损伤幼猪肺表面活性物质的影响

Objective To investigate the effect of recruitment maneuver(RM) on surfactant proteins in young piglets with acute lung injury and the possible mechanisms of lung recruitment after RM.Methods The piglet model of ALI was established by lipopolysaccharide intravenous injection,12 male piglets were randomly divided into two groups:conventional ventilation group(control group) and RM with low tidal volume group(RM group).After 8 hours of ventilation,mRNA expression of surfactant protein-A(SP-A),SP-B,SP-C,SP-D in the piglet lungs were determined by real time PCR and SP-A protein distribution was assessed by immunohistochemistry.Biochemical analyses of TP,total phospholipids(TPL),DSPC were conducted as well.SP-A levels in bronchoalveolar lavage fluid(BALF) and plasma were measured by enzymelinked immunosorbent assay(ELISA).Results As compared with control group,RM group had higher expression of SP-A,SP-B,SP-C and SP-D.SP-A average gray values of control group and RM group were 97.8±6.4 and 106.3±8.5,and there was significant difference(P＜0.01).RM group showed significant increase of TPL,DSPC and DSPC/TP.The concentration of SP-A in BALF was higher in RM group than that of the control group,however,SP-A plasma level was lower in RM group than that of the control group.Conclusion RM can increase suffactant protein expression in ALI animals,alleviate surfactant protein dysfunction and regulate the concentration of SP-A,which may improve alveolar recruitment following the RM and alleviate ventilator-induced lung injury.     

内源性一氧化碳在哮喘中的变化与机制

目的　 探讨内源性一氧化碳 (CO)在哮喘中的变化及其调节机制。 方法 　 5 0只豚鼠随机分成 5组 ,建立哮喘豚鼠模型 ,其中 3组分别使用地米松、血红素氧合酶 1(HO- 1 )特异性激动剂血晶素和抑制剂锡原卟啉 ,余为哮喘组和正常对照组。用分光光度法检测血一氧化碳血红蛋白 (COHb)含量和肺组织HO- 1 活性 ,免疫组化染色观察肺组织HO- 1 蛋白表达等。 结果 　哮喘组全血COHb为 4 94± 2 15 % ;肺HO- 1 活性为 881± 36 1pmol/(mg .protein·h) ,分别较正常组有显著性差异 (t=4 5 8～ 10 19,P <0 0 1) ,肺HO- 1 蛋白增加。血晶素激动组较哮喘组略高 (P >0 0 5 )。锡原卟啉抑制和地塞米松预防组较哮喘组明显降低 (t =4 43～ 8 83,P <0 0 1)。血COHb与肺组织HO- 1 活性水平呈正相关 (r =0 90 ,P <0 0 0 1)。 结论 　哮喘时体内HO- 1 蛋白和活性增加 ,导致内源性CO水平升高     

血管活性肠肽对哮喘小鼠气道炎症及Th17/Treg平衡的影响

目的探讨血管活性肠肽(VIP)对哮喘小鼠气道炎症的影响,以及对Th17细胞/调节性T细胞(Treg)失衡的调控作用。方法将30只BALB/c小鼠随机分成对照组、哮喘组、VIP组,每组10只。利用卵白蛋白(OVA)致敏和激发制作急性哮喘小鼠模型;对照组致敏和激发阶段均以生理盐水代替OVA;VIP组每次OVA激发前,先以VIP溶液(20μg/m L)雾化吸入30 min。留取各组小鼠支气管肺泡灌洗液、肺组织等标本。利用苏木精-伊红染色观察肺组织病理变化;ELISA法检测肺泡灌洗液中Th17/Treg相关细胞因子水平;免疫组化及Real-time PCR检测肺组织中Th17细胞特异性转录因子RORγt及Treg特异性转录因子Foxp3表达情况。结果病理组织学结果显示VIP组小鼠肺组织气道炎症表现较哮喘组减轻。哮喘组小鼠BALF中IL-17浓度高于对照组(P0.01),VIP组IL-17浓度较哮喘组降低(P0.01),但仍高于对照组(P0.01)。哮喘组BALF中IL-10浓度低于对照组(P0.01),VIP组IL-10浓度较哮喘组升高(P0.01),但仍低于对照组(P0.01)。哮喘组小鼠肺组织RORγt mRNA及蛋白表达高于对照组(P0.01),Foxp3 mRNA及蛋白表达低于对照组(P0.01);VIP组肺组织RORγt mRNA及蛋白表达水平低于哮喘组(P0.01),Foxp3 mRNA及蛋白表达水平高于哮喘组(P0.05)。结论哮喘小鼠存在Th17/Treg免疫失衡,VIP可通过调控Th17/Treg免疫失衡而改善哮喘小鼠气道炎症。     

血红素类物质与哮喘发作关系的初步探讨

目的　探讨血红素类物质在哮喘中的变化及与哮喘发作的关系。方法　 6 0只豚鼠随机分为正常对照组、哮喘、哮喘自然缓解、地塞米松、血红素氧合酶 1(HO-1)特异性激动剂血晶素和抑制剂锡原卟啉 6组。每组均检测肺组织病理和HO-1的蛋白表达 (免疫组化染色法 ) ,应用分光光度法检测肺组织HO-1活性、血一氧化碳血红蛋白 (COHb)和NO含量 ,放射免疫竞争抑制法测定肺环磷酸鸟苷 (cGMP)。结果　哮喘组和血晶素组①肺HO-1活性 ;②血COHb ;③NO ;④肺cGMP含量与正常组比较 ,差异非常显著 (t值分别为① 5 .6 9,9.2 9,② 6 .2 8,10 .19,③ 5 .77,8.92 ,④ 9.74,6 .96 ,P <0 .0 1) ,血晶素组更为显著。肺HO-1活性 :每小时 (144 9± 42 6 ) pmol/mg ;血COHb (7.43± 2 .0 7) % ;血NO(90 .9± 16 .7) μmol/L ;肺cGMP :(1.96± 0 .6 5 ) pmol/mg ;肺有明显的嗜酸细胞浸润 ,HO-1蛋白表达≥ 4级。锡原卟啉抑制、地塞米松防治和哮喘自然缓解组各项检测指标显著下降 ,与哮喘组比较差异显著 (P <0 .0 1)。结论　哮喘时 ,体内血红素代谢产物CO和NO、降解血红素的HO-1酶类及其血红素酶类增加 ;血红素类物质增加 ,可能参与哮喘气道高反应性。     

溴化结构域蛋白抑制剂JQ1对哮喘小鼠气道重塑作用机制研究

目的探讨溴化结构域蛋白抑制剂JQ1对哮喘小鼠气道重塑治疗的分子学机制。方法将24只小鼠随机分成对照组、OVA诱导哮喘组(OVA组)、JQ1干预哮喘组(JQ1+OVA组)(n=8)。采用OVA致敏/激发制备哮喘小鼠模型,雾化激发前1h,JQ1+OVA组小鼠经腹腔注射JQ1溶液(50μg/g)。末次激发24h后留取支气管肺泡灌洗液(BALF)及肺组织,计算各组BALF中细胞总数及嗜酸性粒细胞百分比,肺组织行病理染色观察各组小鼠肺组织病理改变;采用RT-PCR及Westernblot法分别检测小鼠肺上皮间质转化(EMT)过程中钙黏蛋白(E-Cadherin)、波形蛋白(vimentin)m RNA及蛋白水平的表达变化。结果与对照组比较,OVA组小鼠气道炎性细胞明显浸润,气道壁增厚,黏液渗出增多;BALF中细胞总数增加,嗜酸性粒细胞百分比增多(P0.01)。与OVA组比较,JQ1+OVA组小鼠气道炎症反应明显减轻;BALF中细胞总数明显减少,嗜酸性粒细胞百分比降低(P0.01)。与对照组比较,OVA组小鼠肺组织气道上皮E-Cadherinm RNA及蛋白表达水平明显下调,vimentinm RNA及蛋白表达水平明显上调(P0.01);与OVA组比较,JQ1+OVA组E-Cadherinm RNA及蛋白表达水平升高,vimentinm RNA及蛋白表达水平下降(P0.01);JQ1+OVA组与对照组上述指标表达水平比较差异无统计学意义(P0.05)。结论 OVA哮喘小鼠存在EMT气道重塑;溴化结构域蛋白抑制剂JQ1可降低哮喘小鼠气道炎症,抑制EMT,减轻气道重塑,为哮喘治疗提供了一个潜在的新方向。     

氯雷他定对哮喘豚鼠白细胞介素5及嗜酸性粒细胞凋亡的影响

目的研究氯雷他定对哮喘豚鼠支气管肺泡灌洗液(BALF)和血清IL-5的表达及肺组织嗜酸性粒细胞(EOS)凋亡延迟状态的影响。方法豚鼠32只随机分为正常对照组、哮喘组、氯雷他定低剂量组、氯雷他定高剂量组。经3次20 g·L-1卵清蛋白(OVA)腹腔注射致敏后,10 g·L-1OVA雾化吸入40 min诱发哮喘发作。氯雷他定低、高剂量组豚鼠分别予2 mg·kg-1·d-1、10mg·kg-1·d-1氯雷他定灌服14 d,每天1次;哮喘组、正常对照组给予9 g·L-1盐水作对照。ELISA法检测其血清和BALF中IL-5水平,罗氏TUNEL原位细胞凋亡检测试剂盒检测其凋亡细胞,HE染色切片随机计数每张切片下200个EOS中凋亡细胞数。结果1.正常对照组、哮喘组、氯雷他定低剂量组、氯雷他定高剂量组血清IL-5水平分别为(0.27±0.04)ng·L-1、(0.41±0.03)ng·L-1、(0.38±0.02)ng·L-1、(0.31±0.03)ng·L-1;BALF中IL-5水平分别为(0.10±0.01)ng·L-1、(0.38±0.04)ng·L-1、(0.33±0.05)ng·L-1、(0.23±0.09)ng·L-1,血清及BALF中IL-5水平4组间两两比较差异均有统计学意义(Pa<0.05)。2.正常对照组、哮喘组、氯雷他定低剂量组、氯雷他定高剂量组豚鼠肺组织EOS发生凋亡的细胞数分别为33个、95个、129个、182个,两两比较差异有统计学意义(χ2=114.5,P<0.05)。其中哮喘组与氯雷他定低剂量组间差异无统计学意义(χ2=10.0,P>0.008 3)。结论氯雷他定对IL-5相关的EOS凋亡延迟状态有抑制作用,能改善肺组织变应性炎症状态,对哮喘的控制具有潜在的积极作用。     

布地奈德对哮喘大鼠模型肺组织血红素氧合酶-1的调控作用

目的：研究大鼠哮喘模型肺组织血红素氧合酶-1(heme oxygenase-1,HO-1)蛋白及基因表达特点,以及布地奈德(budesonide,BUD)对HO-1蛋白及基因表达的影响。方法：用卵清蛋白(OVA)致敏、激发大鼠建立哮喘动物模型,并在致敏、激发过程中分别经地塞米松(DXM) 、血晶素(Hemin)及BUD处理。分别测定激发后各组动物全血COHb 的百分比含量;计算肺泡灌洗液（BALF）沉渣中细胞总数和分类百分比;进行支气管周围炎性细胞浸润评分;利用免疫组化和RT-PCR检测方式观察HO-1在哮喘大鼠肺组织的蛋白及基因表达。结果：肺组织的病理改变及BALF细胞学检测显示Hemin (H)组、DXM (D)组和BUD (B)组炎性细胞浸润较哮喘(A)组有显著减轻(P<0.01或P<0.05）。与对照(C)组相比,A、H、D和B组HO-1蛋白、基因表达以及碳氧血红蛋白(COHb)含量显著性升高(P<0.01);而与A组相比,H、D和B组HO-1蛋白及mRNA表达亦显著升高(P<0.01或P<0.05）;D组和H组COHb含量显著升高（P<0.05）。结论：哮喘大鼠的肺组织HO-1表达水平及活性显著增加,提示HO-1可能参与了哮喘的发病过程;HO-1对大鼠哮喘模型气道炎症有保护作用;BUD和DXM对大鼠哮喘模型气道炎症有保护作用,可能通过上调肺组织HO-1表达实现。     

褪黑素对哮喘大鼠肺组织活性氧产生的影响(英文)

目的　支气管哮喘是慢性气道炎症性疾病 ,肺组织活性氧产生可以导致气道炎症 ,本研究目的探讨褪黑素 (MT)对哮喘模型大鼠肺组织活性氧 (ROS)生成以及气道炎症的影响。方法　将 2 4只大鼠随机分为 3组 :哮喘组 (n=8) :用 10 %鸡卵白蛋白 (OVA) 1ml、氢氧化铝凝胶 10 0mg无菌腹腔注入 ,2周后用 1%OVA超声雾化吸入 2 0min ,连续激发 1周致其哮喘发作 ;MT组 (n =8) :模型制作同哮喘组 ,在每次激发前 30min腹腔注入MT 10mg/kg ;对照组 (n =8) :以生理盐水代替OVA吸入。每组分别于最后一次激发后 6h测定气道反应性 ;取支气管肺泡灌洗液 (BALF)进行白细胞计数、分类 ;取肺组织进行活性氧产量测定。结果　MT组大鼠气道反应性及BALF中炎性细胞数明显低于哮喘组 ,差异有显著性 (P <0 .0 5 )。肺组织活性氧产量在哮喘组、MT组及对照组分别为 (114 .8± 11.3)U/mgprot、(95 .2± 5 .9)U/mgprot及 (87.5± 7.4 )U/mgprot,哮喘组高于其它两组 ,差异均有显著性 (P <0 .0 5 )。结论　哮喘组大鼠肺组织活性氧产量增加。MT干预可以降低肺组织活性氧产生 ,降低气道炎症和气道高反应性 ,这可能是其治疗哮喘的保护机制。     

瞬时电位感受器香草酸受体1在哮喘小鼠气道炎症中的作用研究

目的观察瞬时电位感受器香草酸受体1(TRPV1)通道活性改变对哮喘模型小鼠气道炎症程度的影响。方法将BALB/c小鼠随机分为对照组、哮喘组、辣椒素(TRPV1激动剂)组、辣椒平(TRPV1拮抗剂)组、地塞米松组。采用卵清蛋白-氢氧化铝混合液腹腔注射致敏并雾化激发构建哮喘小鼠模型。辣椒素、辣椒平和地塞米松组分别在每次激发前30min腹腔注射辣椒素(30μg/kg)、辣椒平(10μmol/kg)及地塞米松(2mg/kg)。采用苏木精-伊红染色观察各组小鼠肺部炎症程度;ELISA法检测肺泡灌洗液(BALF)中IL-8和IL-13的含量;Real-Time PCR法检测小鼠肺组织中TRPV1m RNA的相对含量。结果与哮喘组比较,辣椒平组和地塞米松组肺部炎症程度减轻,辣椒素组炎症程度明显加重。与对照组比较,哮喘组、辣椒素组小鼠BALF中IL-13、IL-8含量及肺组织TRPV1m RNA表达明显增加(P0.05);与哮喘组比较,辣椒平组、地塞米松组BALF中IL-13、IL-8含量及肺组织TRPV1m RNA表达明显降低(P0.05),辣椒素组BALF中IL-13、IL-8含量明显增加(P0.05)。结论 TRPV1通道激活剂和通道抑制剂可影响哮喘小鼠肺部炎症程度。地塞米松可能通过调节TRPV1水平减轻气道炎症。     

mTOR/4EBP1/HIF-1α/VEGF信号通路在哮喘小鼠肺组织中的表达及意义

目的探讨哺乳动物雷帕霉素靶蛋白(m TOR)/真核生物始动因子4E结合蛋白1(4EBP1)/缺氧诱导因子-1α(HIF-1α)/血管内皮生长因子(VEGF)信号通路在哮喘小鼠中的表达及意义。方法 40只SPF级6~8周龄雌性Balb/c小鼠随机分为对照组、哮喘组、布地奈德干预组及m TOR抑制剂(雷帕霉素)干预组,每组10只。卵清蛋白致敏激发建立哮喘小鼠模型,各干预组分别在激发前30 min给予雷帕霉素3 mg/kg腹腔注射或布地奈德混悬液1 mg雾化吸入,对照组和哮喘组以生理盐水代替。于末次激发24 h后处死小鼠,收集肺泡灌洗液(BALF),采用ELISA法测定HIF-1α、VEGF水平;取肺组织行苏木精-伊红(HE)染色观察其病理变化;免疫组化染色和Western blot法测定肺组织磷酸化的m TOR及4EBP1(p-m TOR及p-4EBP1)蛋白表达水平。Pearson法分析p-m TOR、p-4EBP1、HIF-1α、VEGF表达的相关性。结果与对照组相比,哮喘组气管及其周围炎性细胞浸润明显,分泌物增多;BALF中HIF-1α、VEGF水平显著升高(P0.01);肺组织p-m TOR、p-4EBP1表达显著增加(P0.01)。与哮喘组相比,各干预组气道炎症浸润明显减轻,分泌物减少;BALF中HIF-1α、VEGF水平明显下降(P0.01);肺组织p-m TOR、p-4EBP1表达显著降低(P0.01)。对照组及两干预组间相比上述指标变化差异均无统计学意义(P0.05)。哮喘组小鼠p-m TOR、p-4EBP1、HIF-1α、VEGF表达水平两两互呈正相关(P0.05),而对照组及两干预组各指标间无相关性(P0.05)。结论哮喘发生时p-m TOR、p-4EBP1、HIF-1α、VEGF可能协同参与了哮喘的发病过程。雷帕霉素能阻断这一过程,可能作为治疗哮喘的新靶点。     

热休克因子1对过敏性哮喘小鼠气道高反应性和气道炎症的影响

目的研究热休克因子1(HSF1)对哮喘小鼠气道高反应性和气道炎症的作用及可能机制。方法 36只小鼠随机分为对照组、哮喘组、HSF1小干扰RNA阴性对照组(siHSF1-NC)和siHSF1组,每组9只。采用卵清蛋白(OVA)致敏和激发建立哮喘模型,siHSF1-NC组和siHSF1组小鼠于激发前分别气管内给予siHSF1-NC和siHSF1。末次激发24 h后采用肺功能仪测定气道反应性;直接计数法计算嗜酸性粒细胞(EOS)的数目;ELISA法检测血清OVA特异性Ig E的含量及肺组织和支气管肺泡灌洗液(BALF)中IL-4、IL-5、IL-13和IFN-γ的表达水平;实时荧光定量PCR检测HSF1 mRNA的表达水平;Western blot法检测HSF1、高迁移率蛋白族1(HMGB1)和磷酸化c-jun氨基末端激酶(p-JNK)的蛋白表达水平。结果与对照组相比,哮喘组HSF1 mRNA和蛋白表达升高(P0.05);与siHSF1-NC组相比,siHSF1组HSF1 mRNA和蛋白表达降低(P0.05),且HSF1缺失导致气道壁增厚、气道高反应性增强、OVA特异性Ig E含量和EOS的数目增加(P0.05)。与siHSF1-NC组相比,siHSF1组小鼠BALF和肺组织中IL-4、IL-5和IL-13水平升高,IFN-γ表达减少(P0.05);HMGB1和p-JNK表达升高(P0.05)。结论 HSF1缺失加重哮喘小鼠的气道高反应性和气道炎症,其机制可能是通过负调控HMGB1和JNK来实现。