汽化吸入全氟化碳对内毒素性急性肺损伤兔表面活性蛋白和基质金属蛋白酶的影响

发布时间：2018-05-06 20:43

  本文选题：急性肺损伤 + 全氟化碳　； 参考：《中国人民解放军军医进修学院》2012年博士论文

【摘要】：目的：经汽化吸入全氟化碳(perfluorocarbon, PFC)治疗内毒素性急性肺损伤(acute lung injury, ALI)动物模型,观察汽化吸入PFC对ALI动物表面活性蛋白和基质金属蛋白酶的影响。 方法：健康雌性新西兰白兔24只(平均体重2.48±0.30kg),随机分为4组：①急性肺损伤汽化吸入PFC治疗组(ALI+PFC组)：通过静脉注射脂多糖(lipopolysaccharide, LPS)(2.5mg/kg)制作急性肺损伤动物模型,然后在常规机械通气(容量控制f:40/min, FiO2:0.6, I:E1:1.5, PEEP5cmH2O)的基础上,经呼吸机吸气管路以湿化器加热PFC的方法给予汽化吸入PFC治疗2小时,然后继续常规机械通气6小时。②急性肺损伤组(ALI组)：制作急性肺损伤模型成功后给予常规机械通气8小时。③汽化吸入PFC组(PFC组)：静脉注射与LPS相同剂量的生理盐水,在常规机械通气基础上,经呼吸机吸气管路以湿化器加热PFC的方法给予汽化吸入PFC治疗2小时,然后继续常规机械通气6小时。④正常对照组(C组)：静脉注射与LPS相同剂量的生理盐水,然后给予常规机械通气治疗8小时。试验期间每隔1小时抽取静脉血2ml和动脉血1ml,试验结束后左肺进行支气管肺泡灌洗,留取右肺组织行病理检查和分子生物学检测。 第一部分：观察各组动物呼吸动力学、血液动力学、血气分析的变化；分别利用光学显微镜和透射电镜观察肺组织病理变化。 第二部分：①利用ELISA、real-time PCR和Wester-Blot技术检测血浆、肺泡灌洗液和肺组织中SP-A、SP-B、SP-C、SP-D的表达情况；②利用real-time PCR和Wester-Blot技术检测ErbB1、ErbB4和磷酸化ErbB1在肺组织中的表达情况。 第三部分：利用ELISA和real-time PCR技术检测MMP-2、MMP-9及其抑制剂TIMP-1、TIMP-2在血浆、肺泡灌洗液和肺组织中表达情况。 结果： 1、ALI+PFC组较ALI组氧合指数升高,气道阻力和病理损伤评分降低。PFC组较C组病理损伤评分降低。 2、①与ALI组比较,ALI+PFC组血浆中表面活性蛋白含量下降,肺泡灌洗液中SP-A、SP-B、SP-C含量增加,肺组织中SP-A、SP-B、SP-C mRNA表达增强。②与ALI组比较,ALI+PFC组肺组织中ErbB1和ErbB4mRNA表达减弱；与C组比较,PFC组肺组织中ErbB1和ErbB4mRNA表达增强。 3、与ALI组比较,ALI+PFC组血浆和肺泡灌洗液MMP-2、MMP-9的含量以及MMP-2/TIMP-2和MMP-9/TIMP-1降低,肺组织中MMP-2、MMP-9mRNA表达减弱。与C组比较,PFC组血浆中MMP-2、MMP-9的含量以及血浆和肺泡灌洗液中MMP-2/TIMP-2和MMP-9/TIMP-1降低。 结论： 1、汽化吸入PFC改善肺损伤可能与促进SP-A、SP-B、SP-C的合成与分泌有关。汽化吸入PFC对SP-D的影响尚无法确定。也无法确定汽化吸入PFC对表面活性蛋白的影响是否与ErbB1和ErbB4有关。 2、抑制MMP-2、MMP-9的合成与分泌可能是汽化吸入PFC减轻急性肺损伤的作用机制之一。
[Abstract]:Objective: To observe the effects of vaporization of PFC on the surface active proteins and matrix metalloproteinases of ALI animals by vaporizing inhaled perfluorocarbon (PFC) in the treatment of acute lung injury (ALI).
Methods: 24 healthy female New Zealand white rabbits (average weight of 2.48 0.30kg) were randomly divided into 4 groups: (1) PFC treatment group (group ALI+PFC) with acute lung injury vaporization inhalation (group ALI+PFC): the animal model of acute lung injury was made by intravenous injection of lipopolysaccharide (2.5mg/kg) (2.5mg/kg), and then in conventional mechanical ventilation (volume control f:40/min, FiO2:0.6, On the basis of I:E1:1.5, PEEP5cmH2O), the inhalation inhalation PFC was given for 2 hours through the humidifier heating through the ventilator suction line. Then, the conventional mechanical ventilation was continued for 6 hours. Secondly, the acute lung injury group (group ALI): after making the acute lung injury model, the conventional mechanical ventilation was given for 8 hours. (3) vaporization inhalation PFC group (group PFC): static Intravenous injection of the same dosage of LPS saline, on the basis of conventional mechanical ventilation, on the basis of conventional mechanical ventilation, the method of heating PFC by the humidifier in the ventilator suction line for 2 hours, and then continuing the conventional mechanical ventilation for 6 hours. 4. Normal control group (group C): intravenous injection of the same dose of physiological saline with LPS, and then given the conventional machinery. The venous blood 2ml and arterial blood 1ml were taken every 1 hours during the test. After the test, the left lung was treated with bronchoalveolar lavage, and the right lung tissue was examined by pathological examination and molecular biological examination.
The first part: observe the changes of respiratory dynamics, hemodynamics and blood gas analysis in each group, and observe the pathological changes of lung tissue by optical microscope and transmission electron microscope respectively.
The second part: (1) the expression of SP-A, SP-B, SP-C, SP-D in plasma, alveolar lavage fluid and lung tissue was detected by ELISA, real-time PCR and Wester-Blot.
The third part: using ELISA and real-time PCR technology to detect the expression of MMP-2, MMP-9 and its inhibitor TIMP-1 and TIMP-2 in plasma, bronchoalveolar lavage fluid and lung tissue.
Result锛，

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