大鼠机械通气相关性肺损伤时蛋白激酶Cδ与细胞焦亡的关系  

Relationship between protein kinase C-delta and pyroptosis during ventilator-induced lung injury in rats

作　　者：张晓[1] 宋作艳[1] 李秋杰 王明山[1] 孙立新[1] 韩伟[2] Zhang Xiao;Song Zuoyan;Li Qiujie;Wang Mingshan;Sun Lixin;Han Wei(Department of Anesthesia,Qingdao Municipal Hospital,Medical College of Qingdao University,Qingdao 266071,China;Department of Respiratory and Critical Medicine,Qingdao Municipal Hospital,Medical College of Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学附属青岛市市立医院麻醉科,266071 [2]青岛大学附属青岛市市立医院呼吸与危重症医学科,266071

出　　处：《中华麻醉学杂志》2020年第5期581-584,共4页Chinese Journal of Anesthesiology

基　　金：青岛市医疗卫生优秀学科带头人项目(VDTR2017Y11)。

摘　　要：目的评价大鼠机械通气相关性肺损伤时蛋白激酶Cδ(PKCδ)与细胞焦亡的关系。方法清洁级健康成年雄性SD大鼠36只,体重200~250 g,采用随机数字表法分为3组(n=12):对照组(C组)、机械通气相关性肺损伤组(VILI组)和PKCδ特异性抑制剂KAI 9803组(K组)。气管插管术后VILI组气管内注射200μl磷酸缓冲盐溶液,K组气管内注射KAI 9803200μg/kg,行机械通气4 h(潮气量40 ml/kg、通气频率60次/min,吸呼比1∶1,吸入氧浓度21%,呼气末正压为0)。于机械通气结束时采集股动脉血样,进行动脉血气分析,记录PaO2。通气结束后麻醉处死大鼠,取肺组织,制备支气管肺泡灌洗液(BALF),光镜下观察肺组织病理学结果并行肺损伤评分,计算肺组织湿重/干重(W/D)比值,采用考马斯亮蓝法测定BALF总蛋白浓度,采用ELISA法测定BALF IL-18和IL-1β浓度,分别采用Western blot法和qRT-PCR法测定肺组织PKCδ、gasdermin D N端片段(GSDMD-N)及其mRNA的表达。结果与C组比较,VILI组和K组肺损伤评分和W/D比值升高,PaO2降低,BALF总蛋白、IL-18和IL-1β浓度升高,肺组织PKCδ、GSDMD-N及其mRNA表达上调(P<0.01);与VILI组比较,K组肺损伤评分和W/D比值降低,PaO2升高,BALF总蛋白、IL-18和IL-1β浓度降低,肺组织PKCδ、GSDMD-N及其mRNA表达下调(P<0.05或0.01)。结论PKCδ可介导细胞焦亡,参与大鼠机械通气相关性肺损伤的病理生理过程。Objective To evaluate the relationship between protein kinase C-delta(PKCδ)and pyroptosis during ventilator-induced lung injury(VILI)in rats.Methods Thirty-six clean-grade healthy adult male Sprague-Dawley rats,weighing 200-250 g,were divided into 3 groups(n=12 each)using a random number table method:control group(group C),group VILI,and VILI plus specific PKCδinhibitor KAI 9803 group(group K).Phosphate buffer solution 200μl was injected through the tracheal tube after intubation in group VILI,and KAI 9803200μg/kg was given instead in group K.The patients were mechanically ventilated(tidal volume 40 ml/kg,respiratory rate 60 breaths/min,inspiratory/expiratory ratio 1∶2,fraction of inspired oxygen 21%,positive end-expiratory pressure 0)for 4 h.Blood samples were taken from the femoral artery at the end of mechanical ventilation for blood gas analysis,and PaO2 was recorded.Animals were sacrificed at the end of ventilation,lung tissues were removed,and bronchoalveolar lavage fluid(BALF)was prepared.The total protein concentrations in BALF were measured by coomassie blue staining,and concentrations of interleukin-18(IL-18)and IL-1βin BALF by enzyme-linked immunosorbent assay.Lung tissues were obtained for microscopic examination of the pathological changes which were scored and for determination of wet/dry weight ratio(W/D ratio)and expression of PKCδand gasdermin D N terminal fragment(GSDMD-N)protein and mRNA(by Western blot or by quantitative real-time polymerase chain reaction).Results Compared with group C,the lung injury score,W/D ratio,and concentrations of total protein,IL-18 and IL-1βin BALF were significantly increased,PaO2 was decreased,and the expression of PKCδand GSDMD-N protein and mRNA was up-regulated in VILI and K groups(P<0.01).Compared with group VILI,the lung injury score,W/D ratio,and concentrations of total protein,IL-18 and IL-1βin BALF were significantly decreased,PaO2 was increased,and the expression of PKCδand GSDMD-N protein and mRNA was down-regulated in group K(P<0.05 or 0.01)

关 键 词：蛋白激酶C-δ 细胞焦亡 呼吸 人工 急性肺损伤 

分 类 号：R563[医药卫生—呼吸系统]