硫氧还蛋白在高氧肺损伤中的作用
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摘要
第一部分高氧对硫氧还蛋白系统表达的影响
【目的】建立早产新生大鼠高氧暴露肺损伤动物模型,观察高浓度氧对早产鼠肺组织形态学影响以及硫氧还蛋白系统表达水平的变化,探讨硫氧还蛋白系统在高氧肺损伤中的可能作用。
【方法】孕21天SD大鼠行剖腹产术,娩出的新生鼠为早产鼠。早产SD大鼠生后d1随机分为两组：空气组和高氧组。高氧组持续暴露于氧浓度85%的氧箱内,空气组置于常压空气中。两组分别于空气或高氧暴露后1、4、7、14d,10%乌拉坦腹腔注射麻醉,提取肺组织总RNA和总蛋白,采取半定量逆转录聚合酶链反应(reverse transcription polymerase chain reaction, RT-PCR)检测硫氧还蛋白和硫氧还蛋白还原酶mRNA表达水平；免疫组织化学方法检测硫氧还蛋白在肺组织中的分布和表达强度；免疫印迹法检测肺组织硫氧还蛋白蛋白水平的动态变化；同时采用HE染色观察肺组织形态学改变,并进行辐射状肺泡计数。
【结果】①肺组织形态学观察：空气对照组呈正常肺发育状态,高氧组肺组织可见明显肺泡炎性改变和肺发育滞后,同时伴有辐射状肺泡计数的显著减少(p<0.05)。②与空气对照组比较,高氧暴露诱导了Trx、TrxR mRNA水平的显著上调,同时Trx蛋白水平也呈现相应的升高(p<0.05)③免疫组织化学染色结果显示,Trx普遍分布于肺组织血管内皮细胞和肺泡上皮细胞胞浆内,高氧暴露组Trx强度较对照组增强。
【结论】高氧暴露诱导肺组织广泛的炎症反应及肺发育落后,同时上调肺组织Trx、TrxR表达水平。Trx系统表达增高可能在早产鼠高氧肺损伤的发病机制中发挥重要保护作用。
第二部分硫氧还蛋白对高氧暴露胎鼠肺泡上皮细胞的保护作用
【目的】观察补充外源性硫氧还蛋白对高氧诱导的胎鼠肺泡Ⅱ型上皮细胞损伤的影响,探讨硫氧还蛋白可能的作用机制。
【方法】无菌分离、培养胎鼠肺泡Ⅱ型上皮细胞。随机将细胞分为单纯高氧暴露组和重组人硫氧还蛋白(recombinant human thioredoxin,rhTrx)处理组,两组同时暴露于氧浓度95%的高氧环境。两组分别于高氧暴露0、12、24、48h,MTT法检测细胞活力情况；流式细胞术检测细胞凋亡和细胞内活性氧(reactive oxygen species,ROS)的生成；超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶活性分别由其相应的酶活性检测试剂盒进行测定；免疫印迹蛋白检测法测定丝裂原激活蛋白激酶(mitogen-activited protein kinase,MAPK)和磷脂酰肌醇(3)激酶-蛋白激酶(phosphoinositide 3-kinase/protein kinase B,PI3K/Akt)信号通路活性。
【结果】与单纯高氧暴露组细胞比较,①rhTrx处理显著减轻了高氧诱导的肺泡上皮细胞的死亡,提高了细胞活力；②rhTrx显著降低了细胞ROS的水平；③rhTrx显著提高了锰型超氧化物歧化酶和谷胱甘肽过氧化物酶的活性；④rhTrx激活了细胞外信号调节激酶1／2(extracellular signal regulated kinase,ERK 1/2)ERK 1/2和P13K活性,而抑制了c-Jun N末端激酶1/2(c-Jun N-terminal protein kinase,JNK1/2)和P38丝裂原活化蛋白激酶(P38mitogen-activited protein kinase,p38)的活性.
【结论】硫氧还蛋白通过减少细胞内ROS的生成、提高抗氧化酶活性、调控MAPKs和PI3K-Akt信号通路对高氧暴露胎肺泡上皮细胞发挥保护作用。
第三部分硫氧还蛋白在高氧暴露早产鼠肺损伤中作用的相关机制研究
【目的】建立早产新生鼠高氧暴露肺损伤动物模型,观察外源性Trx对高氧暴露早产鼠肺损伤的作用,并探讨Trx的可能作用机制,为高氧肺损伤的防治寻找新的靶点。
【方法】孕21天SD大鼠行剖腹产术,娩出的新生鼠为早产鼠。早产鼠生后dl随机分为3组：①空气对照组,简称空气组；②单纯高氧暴露组,简称高氧组；③Trx干预+高氧暴露组,简称Trx干预组。空气对照组置于室温空气中；单纯高氧暴露组和Trx干预组置于密闭氧箱中,氧浓度维持85%,由数字测氧仪连续监测。Trx干预组每日腹腔注射rhTrx (2mg/kg),空气组和单纯高氧暴露组注射与rhTrx同体积的PBS。每日观察早产鼠生长状态、定时监测体重、记录死亡情况。于空气或高氧暴露后14d,取肺组织标本分别称量肺湿和干重并计算湿干比；石蜡切片行HE染色观察肺组织的病理学变化并进行辐射状肺泡计数(radical alveolar counts,RAC)；荧光实时定量PCR (real-time PCR)技术检测肺组织血管内皮生长因子(vascular endothelial growth factor, VEGF)、血管内皮生长因子受体(vascular endothelial growth factor receptor, VEGFR)和Toll样受体(Toll like receptor, TLR) 2、4 mRNA表达变化；免疫印迹蛋白检测技术观察MAPKs、PI3K-Akt信号通路活化情况及caspase 3蛋白水平。
【结果】①空气对照组未出现早产鼠死亡；高氧暴露早产鼠死亡率较高达28.6%,存活者出现明显生活能力下降和体重增长迟缓；与单纯高氧暴露组比较,rhTrx处理显著提高了早产鼠生存率,死亡率降低到11.1%,生活能力下降和体重增长迟缓情况亦明显改善；②高氧暴露能引起早产鼠肺组织明显炎症反应和肺发育受阻,同时伴有RAC值下降,而应用rhTrx能显著减轻高氧导致的肺损伤形态学改变；③与单纯高氧组比较,rhTrx显著减轻了高氧诱导的早产鼠肺组织水肿；④荧光定量PCR结果：与空气对照组比较,高氧暴露显著抑制早产鼠肺组织VEGF、VEGR mRNA表达,而对TLR4 mRNA具有上调作用,对TLR2无显著影响；rhTrx处理显著提高高氧暴露后肺组织VEGF、VEGR及TLR4基因水平的表达；⑤与单纯高氧暴露组比较,Trx处理组早产鼠肺组织细胞外信号调节激酶1／2(extracellular signal regulated kinase, ERK1/2)和Akt活性显著增高,c-JunN末端激酶1/2 (c-Jun N-terminal protein kinase, JNK1/2)、P38丝裂原活化蛋白激酶(P38mitogen-activited protein kinase, p38)及caspase 3勺活性明显下降(p<0.05)
[结论]硫氧还蛋白在早产鼠高氧肺损伤中具有保护作用；通过诱导VEGF及其受体、TLR4基因水平高表达,调控MAPKs和PI3K-Akt信号通路是其发挥保护作用的具体机制。
Part I The Effect of Hyperoxia on the Expression of Thioredoxin System
[Objective] To set up the animal model of hyperoxic lung injury and investigate the expression levels of thioredoxin system.
[Methods] In the first day after delivery, the preterm SD rats were randomly divided into two groups:Air group and Hyperxia group. The rats in hyperoxia group were exposed to 85% oxygen and those in air group were exposed to room air. The rats were killed in 1, 4,7 and 14 days respectively. Total RNA of lung was isolated and Trx and TrxR mRNA expression levels were detected by reverse transcription polymerase chain reaction (RT-PCR). Immunohistochemistry was used on lung sections to detect the distribution and expression of thioredoxin. Western blot was used to detect the expression of thioredoxin protein level in lung tissue. The sections of lung were stained with HE in order to assess lung histologic changes and examine lung radical alveolar counts (RAC).
[Results]①Rats in hyperxia group showed typical lung injury, which was characterized by alveolitis and delay of lung development. Compared with air group RACs were significantly decreased in hyperxia group (p<0.05).②RT-PCR results showed Trx and TrxR mRNA expression levels in Hyperxia group were increased markedly, and Trx protein level was increased correspondingly.③Immunohistochemistry detected that Trx expressed generally in the cytoplasm of alveolar epithelial cells and vascular endothelial cells.
[Conclusions] Hyperoxia induced the typical change of bronchopulmonary dysplasia. Trx and TrxR expression levels could be induced by hyperoxia, which might play importantly protective role in the development of hyperxia-induced lung injury.
PartⅡRecombinant Human Thioredoxin Protects Fetal TypeⅡEpithelial Cells from Hyperoxia-induced Injury
[Objective] The aim of this study was to determine the role of Trx in the pathogenesis of hyperoxia-induced alveolar epithelial cell injury.
[Methods] Alveolar typeⅡepithelial cells from fetal rat lung were exposed to hyperoxia in vitro in the presence or absence of recombinant human Trx. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Apoptosis and levels of reactive oxygen species (ROS) were measured by flow cytometry. Activation of mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase-Akt (PI3K-Akt) pathways were detected by Western blotting. We also investigated the effects of rhTrx on the following antioxidants (superoxide dismutase, catalase, glutathione peroxidase).
[Results] Trx significantly reduced hyperoxia-induced cell death and increased cell viability. In addition, ROS generation in typeⅡcells was inhibited by rhTrx under hyperoxic conditions. We demonstrated that rhTrx protected typeⅡcells against hyperoxic injury via sustaining the extracellular signal regulated kinase and PI3K activation, and decreasing of c-Jun N-terminal protein kinase and p38 activation. The results also showed manganese superoxide dismutase and glutathione peroxidase activities were increased by rhTrx in typeⅡcells exposed to hyperoxia.
[Conclusions] Taken together, these results demonstrate that rhTrx administration markedly attenuates hyperoxia-induced typeⅡcell injury through reduction of ROS generation, elevation of antioxidant activities and regulation of both MAPK and PI3K-Akt signaling pathways.
Part III Research on the Mechanism of Thioredoxin Protects Preterm Rat from Hyperoxia-induced Lung Injury
[Objective] To set up the animal model of hyperoxic lung injury, investigate the effect of the recombinant human thioredoxin (rhTrx) on hyperoxic lung injury and explore it's mechanism of action.
[Methods] Preterm Sprague-Dawley rats delivered by hysterotomy at 21-d gestation were allowed to recover in room air for 24 h. After one day of birth, pups were randomized to three groups. Group 1 pups were kept in room air and served as controls. Group 2 and 3 pups were oxygen exposed animals maintained in a transparent Plexiglas chambers with 85% oxygen, in which the oxygen concentration was monitored regularly with an oxygen sensor. Pups in group 2 received daily an intraperitoneal injection of 2 mg/kg/d of rhTrx, whereas pups in groups 1 and 3 received only its vehicle (PBS). The body weight of all pups was measured every day. All lung tissues of premature rat pups were collected at 14d after birth. The sections of lung were stained with HE in order to assess lung histologic changes and examine lung radical alveolar counts (RAC). The levels of VEGF、VEGFR、TLR2 and TLR4 mRNA were detected by real-time reverse transcription polymerase chain reaction (RT-PCR). The protein expression levels of p-ERK 1/2、ERK1/2、p-JNK1/2、JNK1/2、p-p38、p38、p-Akt、Akt and caspase 3 were determined by western blot.
[Results]①The pups in air group did not dead; the mortality of pups in hyperoxia was 28.6%; compared with hyperoxia group rhTrx pretreatment can decreased the mortality to 11.1%.②Rats in hyperxia group showed typical lung injury, which was characterized by alveolitis and delay of lung development. Compared with air group RACs were significantly decreased in hyperxia group. rhTrx pretreatment improved the hyperoxia induced changes of lung morphometry.③Real-time PCR results showed:exposure to hyperoxia resulted in the levels of mRNA for VEGF and VEGFR were decreased, TLR4 was increased, but TLR2 mRNA expression level did not change. Rat pups treated with rhTrx from the hyperoxic environment expressed significantly high levels of mRNA for VEGF、VEGFR and TLR4 than the hyperoxic control pups.④Wssren blot results showed: the levels of active ERK1/2, JNK1/2, p38 and caspase 3 after exposure to hyperoxia were higher than air control group, and Akt activity level was decreased.The levels of active JNK1/2, p38 and caspase 3 were decreased markedly after rhTrx treatment in hyperoxia exposure rat pups, but active ERK1/2 and Akt were increased with the same rhTrx treatment.
[Conclusions] Trx had a protective effect on hyperoxic lung injury by which decrease active levels of JNK, p38 and caspase 3, increase active levels of ERK and Akt, subsequently increase the expression of VEGF, VEGFR and TRL4.
【目的】建立早产新生大鼠高氧暴露肺损伤动物模型,观察高浓度氧对早产鼠肺组织形态学影响以及硫氧还蛋白系统表达水平的变化,探讨硫氧还蛋白系统在高氧肺损伤中的可能作用。
【方法】孕21天SD大鼠行剖腹产术,娩出的新生鼠为早产鼠。早产SD大鼠生后d1随机分为两组：空气组和高氧组。高氧组持续暴露于氧浓度85%的氧箱内,空气组置于常压空气中。两组分别于空气或高氧暴露后1、4、7、14d,10%乌拉坦腹腔注射麻醉,提取肺组织总RNA和总蛋白,采取半定量逆转录聚合酶链反应(reverse transcription polymerase chain reaction, RT-PCR)检测硫氧还蛋白和硫氧还蛋白还原酶mRNA表达水平；免疫组织化学方法检测硫氧还蛋白在肺组织中的分布和表达强度；免疫印迹法检测肺组织硫氧还蛋白蛋白水平的动态变化；同时采用HE染色观察肺组织形态学改变,并进行辐射状肺泡计数。
【结果】①肺组织形态学观察：空气对照组呈正常肺发育状态,高氧组肺组织可见明显肺泡炎性改变和肺发育滞后,同时伴有辐射状肺泡计数的显著减少(p<0.05)。②与空气对照组比较,高氧暴露诱导了Trx、TrxR mRNA水平的显著上调,同时Trx蛋白水平也呈现相应的升高(p<0.05)③免疫组织化学染色结果显示,Trx普遍分布于肺组织血管内皮细胞和肺泡上皮细胞胞浆内,高氧暴露组Trx强度较对照组增强。
【结论】高氧暴露诱导肺组织广泛的炎症反应及肺发育落后,同时上调肺组织Trx、TrxR表达水平。Trx系统表达增高可能在早产鼠高氧肺损伤的发病机制中发挥重要保护作用。
第二部分硫氧还蛋白对高氧暴露胎鼠肺泡上皮细胞的保护作用
【目的】观察补充外源性硫氧还蛋白对高氧诱导的胎鼠肺泡Ⅱ型上皮细胞损伤的影响,探讨硫氧还蛋白可能的作用机制。
【方法】无菌分离、培养胎鼠肺泡Ⅱ型上皮细胞。随机将细胞分为单纯高氧暴露组和重组人硫氧还蛋白(recombinant human thioredoxin,rhTrx)处理组,两组同时暴露于氧浓度95%的高氧环境。两组分别于高氧暴露0、12、24、48h,MTT法检测细胞活力情况；流式细胞术检测细胞凋亡和细胞内活性氧(reactive oxygen species,ROS)的生成；超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶活性分别由其相应的酶活性检测试剂盒进行测定；免疫印迹蛋白检测法测定丝裂原激活蛋白激酶(mitogen-activited protein kinase,MAPK)和磷脂酰肌醇(3)激酶-蛋白激酶(phosphoinositide 3-kinase/protein kinase B,PI3K/Akt)信号通路活性。
【结果】与单纯高氧暴露组细胞比较,①rhTrx处理显著减轻了高氧诱导的肺泡上皮细胞的死亡,提高了细胞活力；②rhTrx显著降低了细胞ROS的水平；③rhTrx显著提高了锰型超氧化物歧化酶和谷胱甘肽过氧化物酶的活性；④rhTrx激活了细胞外信号调节激酶1／2(extracellular signal regulated kinase,ERK 1/2)ERK 1/2和P13K活性,而抑制了c-Jun N末端激酶1/2(c-Jun N-terminal protein kinase,JNK1/2)和P38丝裂原活化蛋白激酶(P38mitogen-activited protein kinase,p38)的活性.
【结论】硫氧还蛋白通过减少细胞内ROS的生成、提高抗氧化酶活性、调控MAPKs和PI3K-Akt信号通路对高氧暴露胎肺泡上皮细胞发挥保护作用。
第三部分硫氧还蛋白在高氧暴露早产鼠肺损伤中作用的相关机制研究
【目的】建立早产新生鼠高氧暴露肺损伤动物模型,观察外源性Trx对高氧暴露早产鼠肺损伤的作用,并探讨Trx的可能作用机制,为高氧肺损伤的防治寻找新的靶点。
【方法】孕21天SD大鼠行剖腹产术,娩出的新生鼠为早产鼠。早产鼠生后dl随机分为3组：①空气对照组,简称空气组；②单纯高氧暴露组,简称高氧组；③Trx干预+高氧暴露组,简称Trx干预组。空气对照组置于室温空气中；单纯高氧暴露组和Trx干预组置于密闭氧箱中,氧浓度维持85%,由数字测氧仪连续监测。Trx干预组每日腹腔注射rhTrx (2mg/kg),空气组和单纯高氧暴露组注射与rhTrx同体积的PBS。每日观察早产鼠生长状态、定时监测体重、记录死亡情况。于空气或高氧暴露后14d,取肺组织标本分别称量肺湿和干重并计算湿干比；石蜡切片行HE染色观察肺组织的病理学变化并进行辐射状肺泡计数(radical alveolar counts,RAC)；荧光实时定量PCR (real-time PCR)技术检测肺组织血管内皮生长因子(vascular endothelial growth factor, VEGF)、血管内皮生长因子受体(vascular endothelial growth factor receptor, VEGFR)和Toll样受体(Toll like receptor, TLR) 2、4 mRNA表达变化；免疫印迹蛋白检测技术观察MAPKs、PI3K-Akt信号通路活化情况及caspase 3蛋白水平。
【结果】①空气对照组未出现早产鼠死亡；高氧暴露早产鼠死亡率较高达28.6%,存活者出现明显生活能力下降和体重增长迟缓；与单纯高氧暴露组比较,rhTrx处理显著提高了早产鼠生存率,死亡率降低到11.1%,生活能力下降和体重增长迟缓情况亦明显改善；②高氧暴露能引起早产鼠肺组织明显炎症反应和肺发育受阻,同时伴有RAC值下降,而应用rhTrx能显著减轻高氧导致的肺损伤形态学改变；③与单纯高氧组比较,rhTrx显著减轻了高氧诱导的早产鼠肺组织水肿；④荧光定量PCR结果：与空气对照组比较,高氧暴露显著抑制早产鼠肺组织VEGF、VEGR mRNA表达,而对TLR4 mRNA具有上调作用,对TLR2无显著影响；rhTrx处理显著提高高氧暴露后肺组织VEGF、VEGR及TLR4基因水平的表达；⑤与单纯高氧暴露组比较,Trx处理组早产鼠肺组织细胞外信号调节激酶1／2(extracellular signal regulated kinase, ERK1/2)和Akt活性显著增高,c-JunN末端激酶1/2 (c-Jun N-terminal protein kinase, JNK1/2)、P38丝裂原活化蛋白激酶(P38mitogen-activited protein kinase, p38)及caspase 3勺活性明显下降(p<0.05)
[结论]硫氧还蛋白在早产鼠高氧肺损伤中具有保护作用；通过诱导VEGF及其受体、TLR4基因水平高表达,调控MAPKs和PI3K-Akt信号通路是其发挥保护作用的具体机制。
Part I The Effect of Hyperoxia on the Expression of Thioredoxin System
[Objective] To set up the animal model of hyperoxic lung injury and investigate the expression levels of thioredoxin system.
[Methods] In the first day after delivery, the preterm SD rats were randomly divided into two groups:Air group and Hyperxia group. The rats in hyperoxia group were exposed to 85% oxygen and those in air group were exposed to room air. The rats were killed in 1, 4,7 and 14 days respectively. Total RNA of lung was isolated and Trx and TrxR mRNA expression levels were detected by reverse transcription polymerase chain reaction (RT-PCR). Immunohistochemistry was used on lung sections to detect the distribution and expression of thioredoxin. Western blot was used to detect the expression of thioredoxin protein level in lung tissue. The sections of lung were stained with HE in order to assess lung histologic changes and examine lung radical alveolar counts (RAC).
[Results]①Rats in hyperxia group showed typical lung injury, which was characterized by alveolitis and delay of lung development. Compared with air group RACs were significantly decreased in hyperxia group (p<0.05).②RT-PCR results showed Trx and TrxR mRNA expression levels in Hyperxia group were increased markedly, and Trx protein level was increased correspondingly.③Immunohistochemistry detected that Trx expressed generally in the cytoplasm of alveolar epithelial cells and vascular endothelial cells.
[Conclusions] Hyperoxia induced the typical change of bronchopulmonary dysplasia. Trx and TrxR expression levels could be induced by hyperoxia, which might play importantly protective role in the development of hyperxia-induced lung injury.
PartⅡRecombinant Human Thioredoxin Protects Fetal TypeⅡEpithelial Cells from Hyperoxia-induced Injury
[Objective] The aim of this study was to determine the role of Trx in the pathogenesis of hyperoxia-induced alveolar epithelial cell injury.
[Methods] Alveolar typeⅡepithelial cells from fetal rat lung were exposed to hyperoxia in vitro in the presence or absence of recombinant human Trx. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Apoptosis and levels of reactive oxygen species (ROS) were measured by flow cytometry. Activation of mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase-Akt (PI3K-Akt) pathways were detected by Western blotting. We also investigated the effects of rhTrx on the following antioxidants (superoxide dismutase, catalase, glutathione peroxidase).
[Results] Trx significantly reduced hyperoxia-induced cell death and increased cell viability. In addition, ROS generation in typeⅡcells was inhibited by rhTrx under hyperoxic conditions. We demonstrated that rhTrx protected typeⅡcells against hyperoxic injury via sustaining the extracellular signal regulated kinase and PI3K activation, and decreasing of c-Jun N-terminal protein kinase and p38 activation. The results also showed manganese superoxide dismutase and glutathione peroxidase activities were increased by rhTrx in typeⅡcells exposed to hyperoxia.
[Conclusions] Taken together, these results demonstrate that rhTrx administration markedly attenuates hyperoxia-induced typeⅡcell injury through reduction of ROS generation, elevation of antioxidant activities and regulation of both MAPK and PI3K-Akt signaling pathways.
Part III Research on the Mechanism of Thioredoxin Protects Preterm Rat from Hyperoxia-induced Lung Injury
[Objective] To set up the animal model of hyperoxic lung injury, investigate the effect of the recombinant human thioredoxin (rhTrx) on hyperoxic lung injury and explore it's mechanism of action.
[Methods] Preterm Sprague-Dawley rats delivered by hysterotomy at 21-d gestation were allowed to recover in room air for 24 h. After one day of birth, pups were randomized to three groups. Group 1 pups were kept in room air and served as controls. Group 2 and 3 pups were oxygen exposed animals maintained in a transparent Plexiglas chambers with 85% oxygen, in which the oxygen concentration was monitored regularly with an oxygen sensor. Pups in group 2 received daily an intraperitoneal injection of 2 mg/kg/d of rhTrx, whereas pups in groups 1 and 3 received only its vehicle (PBS). The body weight of all pups was measured every day. All lung tissues of premature rat pups were collected at 14d after birth. The sections of lung were stained with HE in order to assess lung histologic changes and examine lung radical alveolar counts (RAC). The levels of VEGF、VEGFR、TLR2 and TLR4 mRNA were detected by real-time reverse transcription polymerase chain reaction (RT-PCR). The protein expression levels of p-ERK 1/2、ERK1/2、p-JNK1/2、JNK1/2、p-p38、p38、p-Akt、Akt and caspase 3 were determined by western blot.
[Results]①The pups in air group did not dead; the mortality of pups in hyperoxia was 28.6%; compared with hyperoxia group rhTrx pretreatment can decreased the mortality to 11.1%.②Rats in hyperxia group showed typical lung injury, which was characterized by alveolitis and delay of lung development. Compared with air group RACs were significantly decreased in hyperxia group. rhTrx pretreatment improved the hyperoxia induced changes of lung morphometry.③Real-time PCR results showed:exposure to hyperoxia resulted in the levels of mRNA for VEGF and VEGFR were decreased, TLR4 was increased, but TLR2 mRNA expression level did not change. Rat pups treated with rhTrx from the hyperoxic environment expressed significantly high levels of mRNA for VEGF、VEGFR and TLR4 than the hyperoxic control pups.④Wssren blot results showed: the levels of active ERK1/2, JNK1/2, p38 and caspase 3 after exposure to hyperoxia were higher than air control group, and Akt activity level was decreased.The levels of active JNK1/2, p38 and caspase 3 were decreased markedly after rhTrx treatment in hyperoxia exposure rat pups, but active ERK1/2 and Akt were increased with the same rhTrx treatment.
[Conclusions] Trx had a protective effect on hyperoxic lung injury by which decrease active levels of JNK, p38 and caspase 3, increase active levels of ERK and Akt, subsequently increase the expression of VEGF, VEGFR and TRL4.
引文
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