文章摘要
γ-氨基丁酸预先给药对机械通气所致人肺泡Ⅱ型上皮细胞损伤的影响
Effect of γ-aminobutyric acid preconditioning on pathological stretch-induced damage to type Ⅱ alveolar epithelial cells by mechanical ventilation
  
DOI：10.12089/jca.2018.01.015
中文关键词: γ-氨基丁酸  呼吸窘迫综合征  机械通气
英文关键词: γ-Aminobutyric acid  Respiratory distress syndrome  Mechanical ventilation
基金项目:国家自然科学基金(81571936, 81601679,81401626); 江苏省“六大人才高峰”(WSW 077)
作者单位E-mail
罗科 225001,扬州大学临床医学院,江苏省苏北人民医院麻醉科  
高巨 225001,扬州大学临床医学院,江苏省苏北人民医院麻醉科 gaoju_003@163.com 
方向志 225001,扬州大学临床医学院,江苏省苏北人民医院麻醉科  
黄天丰 225001,扬州大学临床医学院,江苏省苏北人民医院麻醉科  
张扬 225001,扬州大学临床医学院,江苏省苏北人民医院麻醉科  
郭唯真 225001,扬州大学临床医学院,江苏省苏北人民医院麻醉科  
葛亚丽 225001,扬州大学临床医学院,江苏省苏北人民医院麻醉科  
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中文摘要:
      目的 评价γ-氨基丁酸(GABA)预先给药对机械通气所致人肺泡Ⅱ型上皮细胞损伤的影响。方法 人肺泡Ⅱ型上皮细胞株A549细胞,以0.2×106/ml接种培养板中,每孔2.5 ml,采用随机数字表法,将培养孔分为三组:正常对照组(C组)、机械通气性牵张组(P组)和GABA预先给药组(G组)。C组常规培养;P组给予20%应变率的机械牵张6 h,牵张频率0.3 Hz,载入波形为正弦波;G组机械牵张前30 min时给予50 μmol/L GABA。采用甲基噻唑蓝法测定细胞活力,采用比色法测定乳酸脱氢酶(LDH)含量,计算LDH释放率;采用免疫荧光法观察纤维状肌动蛋白(F-actin)重构情况;采用Western blot法测定RhoA激酶1(ROCK1)与γ 氨基丁酸A型受体(GABAA受体)的相对含量。结果 与C组比较,P组和G组细胞活力明显降低,LDH释放率明显升高,ROCK1的相对含量明显上调,GABAA受体的相对含量明显下调(P<0.05)。与P组比较,G组细胞活力明显升高,LDH释放率明显降低,ROCK1的相对含量明显下调,GABAA受体的相对含量明显上调 (P<0.05)。G组F-actin重构程度轻于P组,而重于C组。结论 GABA预先给药可减轻机械通气性牵张致人肺泡Ⅱ型上皮细胞损伤,其机制可能与GABAA受体的激活有关。
英文摘要:
      Objective To evaluate the effect of γ-aminobutyric acid (GABA) preconditioning on pathological stretch-induced damage to type Ⅱ alveolar epithelial cells by mechanical ventilation. Methods A549 cells cultured in vitro (0.2×106/ml, 2.5 ml/well) were randomly divided into 3 groups using a random number table: control group (group C), pathological stretch group (group P) and GABA preconditioning+pathological stretch group (group G). A549 cells were exposed to 20% cyclic stretch at 0.3 Hz for 6 h in groups P and G; GABA 50 μmol/L was given 30 min before cyclic stretch in group G. After the end of pathological stretch, the cells were collected for determination of the cell viability by methyl thiazolyl tetrazolium assay and lactate dehydrogenase (LDH) release by colorimetricmethod; the expression of F-actin was observed with indirect immunofluorescence; the expression of Rho-associated kinase 1 (ROCK1) and GABAAR were determined by western blot. Results Compared with group C, the cell viability was significantly decreased,the amount of LDH released was increased, the expression of ROCK1 was significantly increased and the expression of GABAAR was significantly decreased in groups P and G (P<0.05); Compared with group P, the cell viability was significantly increased, the amount of LDH released was decreased, F-actin was reconstructed, the expression of ROCK1 was significantly decreased and the expression of GABAAR was significantly increased (P<0.05) in group G. The reconstruction of F-actin in group P was better than that in group G and worse than that in group C. Conclusion GABA preconditioning can attenuate pathological stretch-induced damage to type Ⅱ alveolar epithelial cells probably through upregulating the expression of GABAA receptor.
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