2018 年第 4 期 第 13 卷

蛋白激酶C和酪氨酸激酶在机械通气相关肺损伤中的作用机制

Mechanisms of protein kinase C and tyrosine kinase in mechanical ventilation-induced lung injury

作者：赵涛郑升法李刚韩永彬吕刚

英文作者：

单位：276826济宁医学院附属山东省日照市人民医院麻醉科

英文单位：

关键词：机械通气相关肺损伤；p120-连环蛋白；蛋白激酶C；酪氨酸激酶

英文关键词：

• 摘要：

• 目的    探讨蛋白激酶C（PKC）和酪氨酸激酶（c-Src）在机械通气相关肺损伤（VILI）中的作用机制。方法    将30只无特定病原体级健康C57BL/6J小鼠采用随机数字表法分为5组：正常小潮气量组（C组），大潮气量组（H组），大潮气量+c-Src抑制剂组（H+P组），大潮气量+PKCα抑制剂组（H+B组），大潮气量+c-Src抑制剂+PKCα抑制剂组（H+P+B组），每组6只。C组气管切开后，进行小潮气量机械通气，潮气量为7 ml/kg，呼吸频率为120次/min，呼气末正压为5 cmH2O（1 cmH2O=0.098 kPa）；H组、H+P组、H+B组、H+P+B组气管切开后，进行大潮气量机械通气，潮气量为20 ml/kg，呼吸频率为40次/min，呼气末正压为0 cmH2O； 5组吸呼比为1∶2，通气时间为4 h。H+P组、H+P+B组于麻醉前1 h肌内注射c-Src抑制剂PP2 1 μg/kg；H+B组、H+P+B组于麻醉前1 h肌内注射PKCα抑制剂BIM 0.12 mg/kg。机械通气结束后，处死小鼠，取肺组织，记录肺损伤病理学评分并测肺组织湿干重比，采用蛋白质印迹法测定p120-连环蛋白、PKCα、c-Src的表达。结果    H组肺损伤病理学评分及肺组织湿干重比高于C组、H+P组、H+B组和H+P+B组，差异均有统计学意义（均P<0.05）。H组p120-连环蛋白表达低于、PKCα和p-c-Src表达高于C组［（0.361±0.040）比（0.818±0.025）、（0.731±0.046）比（0.324±0.032）、（0.655±0.040）比（0.207±0.045）］，差异均有统计学意义（均P<0.05）；H+P组、H+B组、H+P+B组p120-连环蛋白表达［（0.714±0.030）、（0.734±0.027）、（0.816±0.020）］明显高于H组，差异均有统计学意义（均P<0.05）；H+B组p-c-Src表达低于H组［（0.268±0.053）比（0.655±0.040）］，差异有统计学意义（P<0.05）。结论    PKCα和c-Src均参与VILI过程，大潮气量机械通气激活PKCα，使c-Src磷酸化，降低p120-连环蛋白表达。

• Objective    To investigate the mechanisms of protein kinase C（PKC） and tyrosine kinase（c-Src） in mechanical ventilation-induced lung injury(VILI). Methods    Thirty healthy specific pathogen free C57BL/6J mice were randomly divided into 5 groups: normal low tidal volume group(group C), high tidal volume group(group H), high tidal volume+c-Src inhibitor group(group H+P), high tidal volume+PKCα inhibitor group(group H+B), high tidal volume+c-Src inhibitor+PKCα inhibitor group(group H+P+B), with 6 mice in each group. Group C was treated by low tidal volume mechanical ventilation: tidal volume＝7 ml/kg, respiratory rate＝120 times/min, end-expiratory positive pressure＝5 cmH2O; group H, group H+P, group H+B and group H+P+B were treated by high tidal volume mechanical ventilation: tidal volume＝20 ml/kg, respiratory rate＝40 times/min, end-expiratory positive pressure＝0 cmH2O; inspiratory-to-expiratory ratio was 1∶2; duration of ventilation was 4 h. Group H+P and group H+P+B had intramuscular injection of c-Src inhibitor PP2（1 μg/kg） 1 h before anesthesia; group H+B and group H+P+B had intramuscular injection of PKCα inhibitor BIM（0.12 mg/kg） 1 h before anesthesia. The mice were executed after mechanical ventilation and lung tissues were separated. Pathological score of lung injury was assessed. Wet-to-dry weight ratio of lung tissue was measured. Expressions of p120-catenin, PKCα and c-Src were tested by Western blot. Results   Pathological score of lung injury and wet-to-dry weight ratio of lung tissue in group H were significantly higher than those in group C, group H+P, group H+B and group H+P+B(P<0.05). Expression of p120-catenin in group H was significantly lower than that in group C, expressions of PKCα and p-c-Src in group H were significantly higher than those in group C［（0.361±0.040） vs （0.818±0.025）, （0.731±0.046） vs （0.324±0.032）, （0.655±0.040） vs （0.207±0.045）］(P<0.05). Expression of p120-catenin in group H+P, group H+B and group H+P+B［（0.714±0.030）,（0.734±0.027）,（0.816±0.020）］ were significantly higher than that in group H(P<0.05). Expression of p-c-Src in group H+B was significantly lower than that in group H［（0.268±0.053）（0.655±0.040）］(P<0.05). Conclusion    sPKCα and c-Src are involved in VILI. High tidal volume mechanical ventilation activates PKCα mediating the phosphorylation of c-Src to decrease the expression of p120-catenin.