七氟醚减轻大鼠呼吸机相关性肺损伤  

Sevoflurane alleviated ventilator-induced lung injury in rats

作　　者：张鲁阳 刘博熙 张晓 秦伟伟 孙立新[1] 李秋杰[1] 韩伟[2] 牛兆倬[3] ZHANG Luyang;LIU Boxi;ZHANG Xiao;QIN Weiwei;SUN Lixin;LI Qiujie;HAN Wei;NIU Zhaozhuo(Department of Anesthesiology,Qingdao Municipal Hospital,Qingdao 266071,China)

机构地区：[1]青岛市市立医院麻醉科,266071 [2]青岛市市立医院呼吸与危重症医学科,266071 [3]青岛市市立医院心脏外科,266071

出　　处：《临床麻醉学杂志》2022年第6期631-637,共7页Journal of Clinical Anesthesiology

摘　　要：目的探讨七氟醚对大鼠呼吸机相关性肺损伤(VILI)的影响并探讨其可能机制。方法健康SPF级雄性SD大鼠36只,6~8周龄,体重220~280 g。随机分为三组:对照组(C组)、VILI组(V组)和七氟醚组(S组),每组12只。大鼠给予1%戊巴比妥钠40 mg/kg麻醉后行气管切开插管术,C组自主呼吸4 h,V组和S组插管后机械通气4 h,S组机械通气期间吸入2%七氟醚4 h。通气参数:V_(T) 20 ml/kg,RR 80次/分,I∶E 1∶1,FiO_(2)21%,PEEP 0 cmH_(2)O。机械通气结束时采集股动脉血测定PaO_(2)。处死大鼠,取肺组织和支气管肺泡灌洗液(BALF),计算肺组织湿/干重比值(W/D),采用ELISA法检测BALF中白细胞介素(IL)-1β、IL-18浓度,二氯荧光黄双乙酸盐法检测BALF中肺泡巨噬细胞活性氧(ROS)水平,Western blot法及qRT-PCR法检测肺组织NF-E2相关因子2(Nrf2)、NLRP3、凋亡相关斑点样蛋白(ASC)、caspase-1蛋白含量及其mRNA表达量,观察肺组织病理学改变并进行肺损伤评分。结果与C组比较,V组和S组机械通气结束时PaO_(2)、肺组织Nrf2蛋白含量及其mRNA表达量明显降低(P<0.05),肺组织W/D、BALF中IL-1β、IL-18浓度、BALF中肺泡巨噬细胞ROS水平、肺组织NLRP3、ASC、caspase-1蛋白含量及其mRNA表达量、肺损伤评分明显升高(P<0.05)。与V组比较,S组机械通气结束时PaO_(2)、肺组织Nrf2蛋白含量及其mRNA表达量明显升高(P<0.05),肺组织W/D、BALF中IL-1β、IL-18浓度、BALF中肺泡巨噬细胞ROS水平、肺组织NLRP3、ASC、caspase-1蛋白含量及其mRNA表达量、肺损伤评分明显降低(P<0.05)。结论七氟醚可能通过上调Nrf2和下调NLRP3炎性小体的表达,减轻肺组织氧化应激反应和炎症反应,减轻大鼠呼吸机相关性肺损伤。Objective To evaluate the effects of sevoflurane on ventilator-induced lung injury(VILI) in rats and its possible mechanism. Methods Thirty-six specific pathogen-free(SPF) healthy adult male Sprague-Dawley rats, aged 6-8 weeks, weighing 220-280 g, were randomly divided into three groups: control group(group C), VILI group(group V) and VILI combined with sevoflurane group(group S), 12 rats in each group. All rats in the three groups were anesthetized with 1% pentobarbital sodium 40 mg/kg and intubated with tracheotomy. Group C maintained spontaneous respiration for 4 hours, group V and group S were treated with mechanical ventilation for 4 hours, group S inhaled 2% sevoflurane for 4 hours during mechanical ventilation. Ventilation parameters were set up as the following: tidal volume 20 ml/kg, respiratory rate 80 breaths/minutes, inspiratory/expiratory ratio 1∶1, fraction of inspired oxygen 21%, positive end-expiratory pressure 0 cmH_(2)O. Blood samples of femoral artery were collected immediately for the detection of PaO_(2)at the end of mechanical ventilation. The rats were sacrificed, then lung tissue samples and bronchoalveolar lavage fluid(BALF) were collected respectively, and wet/dry weight ratio(W/D) of lung tissue was calculated. The concentrations of interleukin(IL)-1β and IL-18 in BALF were determined by enzyme-linked immunosorbent assay(ELISA). The level of reactive oxygen species(ROS) in alveolar macrophages in BALF was detected by DCFH-DA. The expression of NF-E2-related factor 2(Nrf2), NOD-like receptor pyrin domain containing 3(NLRP3), apoptosis-associated speck-like protein containing a CARD(ASC), and caspase-1 protein and mRNA in lung tissues were detected by Western blot and qRT-PCR. Lung histopathological changes were observed and lung injury scores were recorded. Results Compared with group C, PaO_(2)at the end of mechanical ventilation and the expression of Nrf2 protein and mRNA in lung tissues in groups V and S were significantly decreased(P < 0.05), W/D of lung tissue, the concentrations

关 键 词：七氟醚 呼吸机相关性肺损伤 NF-E2相关因子2 NOD样受体热蛋白结构域相关蛋白3 氧化应激 炎症 大鼠 

分 类 号：R614[医药卫生—麻醉学]