雷帕霉素对大鼠呼吸机相关性肺损伤时NLRC4炎症小体活性的影响  

Effect of rapamycin on activity of NLRC4 inflammasomes in rats with ventilator-induced lung injury

作　　者：陈令楠 宋作艳 马福国[1] 李秋杰[1] 孙立新[1] 王明山[1] 韩伟[2] Chen Lingnan;Song Zuoyan;Ma Fuguo;Li Qiujie;Sun Lixin;Wang Mingshan;Han Wei(Department of Anesthesiology,Qingdao Municipal Hospital,Qingdao 266071,China;Department of Respiratory and Critical Care Medicine,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

出　　处：《中华麻醉学杂志》2020年第8期950-954,共5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：目的评价雷帕霉素对大鼠呼吸机相关性肺损伤(VILI)时NOD样受体C4(NLRC4)炎症小体活性的影响。方法清洁级健康雄性SD大鼠36只,6~8周龄,体重200~250 g,采用随机数字表法分为3组(n=12):对照组(C组)、VILI组和雷帕霉素组(RAPA组)。RAPA组造模前连续3 d腹腔注射雷帕霉素4 mg·kg^-1·d^-1,C组和VILI组给予等容量生理盐水。VILI组和RAPA组行机械通气4 h,通气参数设置:RR 80次/min,FiO221%,PEEP 0 cmH2O,I∶E1∶1,VT 20 ml/kg。机械通气结束后采集股动脉血样行血气分析,记录PaO2;采用ELISA法检测血清IL-1β和IL-18浓度。收集左肺支气管肺泡灌洗液(BALF),采用ELISA法测定中性粒细胞计数及IL-1β、IL-18浓度。取肺组织,HE染色后光镜下观察病理学结果,并进行肺损伤评分,测定湿重/干重(W/D)比值,采用Western blot法检测哺乳动物雷帕霉素靶蛋白(mTOR)、NLRC4和caspase-1的表达,采用RT-PCR法检测NLRC4 mRNA表达。结果与C组比较,VILI组和RAPA组肺组织W/D比值、肺损伤评分、BALF中性粒细胞计数及血清、BALF IL-1β、IL-18浓度均升高,PaO2下降,肺组织mTOR、NLRC4、caspase-1及NLRC4 mRNA表达上调(P<0.01);与VILI组比较,RAPA组肺组织W/D比值、肺损伤评分、BALF中性粒细胞计数及血清、BALF IL-1β、IL-18浓度均降低,PaO2升高,肺组织mTOR、NLRC4、caspase-1及NLRC4 mRNA表达下调(P<0.05)。结论雷帕霉素减轻大鼠VILI的机制可能与抑制mTOR信号通路激活,进而抑制NLRC4炎症小体活性有关。Objective To evaluate the effect of rapamycin on the activity of NOD-like receptor C4(NLRC4)inflammasomes in the rats with ventilator-induced lung injury(VILI).Methods Thirty-six healthy clean-grade male Sprague-Dawley rats,aged 6-8 weeks,weighing 200-250 g,were divided into 3 groups(n=12 each)using a random number table method:control group(group C),VILI group and rapamycin group(group RAPA).In group RAPA,rapamycin 4 mg·kg^-1·d^-1 was intraperitoneally injected once a day for 3 consecutive days before establishing the model,while the equal volume of normal saline was given instead in group C and group VILI.The patients were mechanically ventilated for 4 h(tidal volume 20 ml/kg,respiratory rate 80 breaths/min,inspiratory/expiratory ratio 1∶1,fraction of inspired oxygen 21%)in VILI and RAPA groups.Blood samples were collected from the femoral artery after the end of ventilation for blood gas analysis and for determination of serum interleukin-1β(IL-1β)and interleukin-18(IL-18)concentrations(by enzyme-linked immunosorbent assay),and PaO2 was recorded.The bronchoalveolar lavage fluid(BALF)was collected for determination of the neutrophil count and IL-1β and IL-18 concentrations by enzyme-linked immunosorbent assay.The lung tissues were obtained for examination of the pathological changes(under the light microscope)after HE staining which were scored and for determination of wet to dry weight(W/D)ratio,and expression of mammalian target of rapamycin(mTOR),NLRC4 and caspase-1(by Western blot)and expression of NLRC4 mRNA(by real-time polymerase chain reaction).Results Compared with group C,the W/D ratio,lung injury score,neutrophil counts in BALF,and concentrations of IL-1βand IL-18 in serum and BALF were significantly increased,PaO2 was decreased,and the expression of mTOR,NLRC4,caspase-1 and NLRC4 mRNA was up-regulated in group VILI and group RAPA(P<0.01).Compared with group VILI,the W/D ratio,lung injury score,neutrophil counts in BALF,and concentrations of IL-1βand IL-18 in serum and BALF were significantl

关 键 词：西罗莫司 呼吸机相关性肺损伤 NLR蛋白质类 

分 类 号：R965[医药卫生—药理学]