ARDS行控制性肺膨胀的分子生物学机制研究
发布时间：2018-11-13 11:30
【摘要】：目的探讨急性呼吸窘迫综合征(ARDS)行控制性肺膨胀(SI)的治疗效果,并观察SI在分子生物学水平上对ARDS的影响,初步探索其可能的分子生物学机制,以期为临床治疗ARDS提供参考依据。 方法20只成年雄性Beagle犬,麻醉后接呼吸机予以基础通气,利用油酸建立犬ARDS模型。稳定30min后随机分成2组。非SI组：予以一次密闭式吸痰,吸痰后不给予ARDS犬控制性肺膨胀。SI组：予以一次密闭式吸痰后立即给予ARDS犬一次控制性肺膨胀,SI采用持续气道正压通气(CPAP)模式,CPAP设置为30cmH2O持续30秒,结束后恢复基础通气。监测吸痰前、吸痰后3min、15min、30min、60min、120min、240min的动脉血气、血流动力学指标、呼吸力学指标等。油酸前、成模后、吸痰后1小时、吸痰后4小时分别用酶联免疫吸附法(ELISA)检测各组ARDS犬血清中TNF-α、IL-1β、IL-6、IL-10、SP-A和HMGB1。吸痰后4小时静脉注射氯化钾处死ARDS犬,取肺脏标本RT-PCR检测肺组织中AQP-1、AQP-5、SP-A、HMGB1、FLIP和IL-6mRNA的表达,并观察其普通病理。 结果SI提高了ARDS犬密闭式吸痰后的动脉氧分压和静态肺顺应性(P0.05),降低了气道阻力(P0.05),并且SI对ARDS犬的血流动力学指标未产生明显的影响。在分子生物学水平,SI降低了ARDS犬血清中促炎细胞因子(TNF-α、IL-1β、IL-6)和抗炎细胞因子(IL-10)的浓度(P0.05),降低了血清SP-A的水平(P0.05)。在肺组织中,SI能增加AQP-1、AQP-5和FLIP mRNA的表达(P0.05),减少IL-6mRNA的表达(P0.05)。而迟发型炎症因子(HMGB1)在血清和肺组织中的水平两组比较均无明显统计学差异。 结论实施SI能安全、有效的改善ARDS的氧合、提高肺顺应性,改善密闭式吸痰后的低氧血症状态。其可能的分子生物学机制为：SI减轻了剪切力对肺泡上皮细胞及血管内皮细胞的机械刺激,抑制了肺组织中炎症介质的mRNA表达,也减少了全身炎症介质的释放。炎症介质分泌的减少,减轻了肺局部炎症反应和肺泡毛细血管屏障的损伤,肺内SP-A丢失减少,肺泡张力降低,从而预防肺泡塌陷。肺泡毛细血管屏障受损减轻,肺组织内AQP-1和AQP-5表达增加,加强了机体清除水肿液的能力,减轻了肺泡和间质的水肿,改善了氧合。炎症细胞因子产生减少,肺组织中FLIP表达增高,抑制了肺血管内皮细胞、肺泡上皮细胞的凋亡。因此SI减少炎症介质分泌、上调水通道蛋白、减少肺内SP-A丢失等引起的上述一系列变化可能最终达到改善肺顺应性和氧合的目的。
[Abstract]:Objective to investigate the therapeutic effect of (ARDS) on (SI), observe the effect of SI on ARDS at molecular level, and explore its possible molecular biological mechanism. In order to provide reference for clinical treatment of ARDS. Methods Twenty adult male Beagle dogs were treated with ventilator after anesthesia and ARDS model was established by oleic acid. After stable 30min, they were randomly divided into two groups. In non-SI group, once closed aspiration was given, and ARDS dog was not given control pulmonary expansion after sucking sputum. SI group: ARDS dog was given once controlled pulmonary expansion immediately after once closed suction. SI was treated with (CPAP) mode of continuous positive airway pressure ventilation. CPAP was set to 30cmH2O for 30 seconds, after which basic ventilation was restored. The arterial blood gas, hemodynamics, respiratory mechanics and so on were monitored before and 3 min after sputum aspiration. TNF- 伪, IL-1 尾, IL-6,IL-10,SP-A and HMGB1. in serum of ARDS dogs in each group were detected by enzyme linked immunosorbent assay (ELISA) before, after modeling, 1 hour after sputum aspiration and 4 hours after sputum aspiration. ARDS dogs were killed by intravenous injection of potassium chloride 4 hours after sputum aspiration. The expression of AQP-1,AQP-5,SP-A,HMGB1,FLIP and IL-6mRNA in lung tissue was detected by RT-PCR and the pathological changes were observed. Results SI increased arterial oxygen pressure and static pulmonary compliance after closed sputum aspiration in ARDS dogs (P0.05), decreased airway resistance (P0.05), and SI had no significant effect on hemodynamics of ARDS dogs. At the level of molecular biology, SI decreased the concentration of TNF- 伪, IL-1 尾, IL-6 and anti-inflammatory cytokines (IL-10) in serum of ARDS dogs (P0.05), and decreased the level of serum SP-A (P0.05). In lung tissue, SI could increase the expression of AQP-1,AQP-5 and FLIP mRNA (P0.05) and decrease the expression of IL-6mRNA (P0.05). There was no significant difference in the level of delayed inflammatory factor (HMGB1) in serum and lung tissue between the two groups. Conclusion SI can effectively improve oxygenation of ARDS, improve lung compliance and improve hypoxemia after closed sputum aspiration. The possible molecular biological mechanism is that SI alleviates the mechanical stimulation of alveolar epithelial cells and vascular endothelial cells by shear stress, inhibits the mRNA expression of inflammatory mediators in lung tissues, and reduces the release of systemic inflammatory mediators. The decreased secretion of inflammatory mediators alleviates the local inflammatory reaction and the injury of alveolar capillary barrier, decreases the loss of SP-A and the decrease of alveolar tension, thus preventing alveolar collapse. The damage of alveolar capillary barrier was alleviated, the expression of AQP-1 and AQP-5 in lung tissue increased, the ability of removing edema fluid was strengthened, the edema of alveolar and interstitial was alleviated, and oxygenation was improved. The production of inflammatory cytokines decreased and the expression of FLIP in lung tissue increased, which inhibited the apoptosis of pulmonary vascular endothelial cells and alveolar epithelial cells. Therefore, SI can reduce the secretion of inflammatory mediators, upregulate aquaporin and decrease the loss of SP-A in the lung, which may ultimately improve lung compliance and oxygenation.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R563.8
本文编号：2328979
[Abstract]:Objective to investigate the therapeutic effect of (ARDS) on (SI), observe the effect of SI on ARDS at molecular level, and explore its possible molecular biological mechanism. In order to provide reference for clinical treatment of ARDS. Methods Twenty adult male Beagle dogs were treated with ventilator after anesthesia and ARDS model was established by oleic acid. After stable 30min, they were randomly divided into two groups. In non-SI group, once closed aspiration was given, and ARDS dog was not given control pulmonary expansion after sucking sputum. SI group: ARDS dog was given once controlled pulmonary expansion immediately after once closed suction. SI was treated with (CPAP) mode of continuous positive airway pressure ventilation. CPAP was set to 30cmH2O for 30 seconds, after which basic ventilation was restored. The arterial blood gas, hemodynamics, respiratory mechanics and so on were monitored before and 3 min after sputum aspiration. TNF- 伪, IL-1 尾, IL-6,IL-10,SP-A and HMGB1. in serum of ARDS dogs in each group were detected by enzyme linked immunosorbent assay (ELISA) before, after modeling, 1 hour after sputum aspiration and 4 hours after sputum aspiration. ARDS dogs were killed by intravenous injection of potassium chloride 4 hours after sputum aspiration. The expression of AQP-1,AQP-5,SP-A,HMGB1,FLIP and IL-6mRNA in lung tissue was detected by RT-PCR and the pathological changes were observed. Results SI increased arterial oxygen pressure and static pulmonary compliance after closed sputum aspiration in ARDS dogs (P0.05), decreased airway resistance (P0.05), and SI had no significant effect on hemodynamics of ARDS dogs. At the level of molecular biology, SI decreased the concentration of TNF- 伪, IL-1 尾, IL-6 and anti-inflammatory cytokines (IL-10) in serum of ARDS dogs (P0.05), and decreased the level of serum SP-A (P0.05). In lung tissue, SI could increase the expression of AQP-1,AQP-5 and FLIP mRNA (P0.05) and decrease the expression of IL-6mRNA (P0.05). There was no significant difference in the level of delayed inflammatory factor (HMGB1) in serum and lung tissue between the two groups. Conclusion SI can effectively improve oxygenation of ARDS, improve lung compliance and improve hypoxemia after closed sputum aspiration. The possible molecular biological mechanism is that SI alleviates the mechanical stimulation of alveolar epithelial cells and vascular endothelial cells by shear stress, inhibits the mRNA expression of inflammatory mediators in lung tissues, and reduces the release of systemic inflammatory mediators. The decreased secretion of inflammatory mediators alleviates the local inflammatory reaction and the injury of alveolar capillary barrier, decreases the loss of SP-A and the decrease of alveolar tension, thus preventing alveolar collapse. The damage of alveolar capillary barrier was alleviated, the expression of AQP-1 and AQP-5 in lung tissue increased, the ability of removing edema fluid was strengthened, the edema of alveolar and interstitial was alleviated, and oxygenation was improved. The production of inflammatory cytokines decreased and the expression of FLIP in lung tissue increased, which inhibited the apoptosis of pulmonary vascular endothelial cells and alveolar epithelial cells. Therefore, SI can reduce the secretion of inflammatory mediators, upregulate aquaporin and decrease the loss of SP-A in the lung, which may ultimately improve lung compliance and oxygenation.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R563.8
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