摘要
目的评价姜黄素减轻脓毒症小鼠急性肺损伤(ALI)时沉默信息调节因子1(SIRT1)与铁死亡的关系。方法SPF级雄性C57BL/6J小鼠152只,8周龄,体质量23~27 g,采用随机数字表法分为4组(n=38):假手术组(C组)、脓毒症组(S组)、姜黄素组(Cur组)和姜黄素+SIRT1抑制剂EX527组(CE组)。Cur组每日姜黄素200 mg/kg灌胃;CE组每日姜黄素200 mg/kg灌胃,并腹腔注射EX5275 mg/kg;C组和S组给予等量溶剂二甲基亚砜。各组连续给药5 d后采用盲肠结扎穿孔术制备脓毒症模型。每组随机取20只小鼠观察造模后7 d生存情况。造模后24 h时,收集支气管肺泡灌洗液(BALF),采用ELISA法检测TNF-α、IL-1β、IL-6和IL-18浓度;取肺组织,计算湿重/干重比值(W/D比值),HE染色后行肺损伤评分,采用比色法检测肺组织还原型谷胱甘肽(GSH)、MDA和铁含量,Western blot法检测肺组织SIRT1、谷胱甘肽过氧化物酶4(GPX4)、酯酰辅酶A合成酶长链家族成员4(ACSL4)的表达。结果与C组相比,S组造模后7 d生存率降低,BALF中TNF-α、IL-1β、IL-6及IL-18浓度、肺组织W/D比值及肺损伤评分升高,肺组织GSH含量降低,MDA和铁含量升高,SIRT1和GPX4表达下调,ACSL4表达上调(P<0.05);与S组相比,Cur组造模后7 d生存率升高,BALF中TNF-α、IL-1β、IL-6及IL-18浓度、肺组织W/D比值及肺损伤评分降低,肺组织GSH含量增加,MDA和铁含量降低,SIRT1和GPX4表达上调,ACSL4表达下调(P<0.05);与Cur组相比,CE组造模后7 d生存率降低,BALF中TNF-α、IL-1β、IL-6及IL-18浓度、肺组织W/D比值及肺损伤评分升高,肺组织GSH含量降低,MDA和铁含量升高,SIRT1和GPX4表达下调,ACSL4表达上调(P<0.05)。结论姜黄素减轻脓毒症小鼠ALI的机制可能与激活SIRT1,进而抑制铁死亡有关。
Objective To evaluate the relationship between silent information regulator 1(SIRT1)and ferroptosis during curcumin-induced reduction of acute lung injury in a mouse model of sepsis.Methods One hundred and fifty-two SPF-grade male C57BL/6J mice,aged 8 weeks,weighing 23-27 g,were divided into 4 groups(n=38 each)using a random number table method:sham operation group(C group),sepsis group(S group),curcumin group(Cur group)and curcumin plus SIRT1 inhibitor EX527 group(CE group).Curcumin 200 mg/kg was administered by intragastric gavage every day in Cur group.Curcumin 200 mg/kg was administered by intragastric gavage every day and EX5275 mg/kg was intraperitoneally injected in CE group.The equal volume of solvent dimethyl sulfoxide(DMSO)was given in C group and S group.Sepsis model was developed by cecal ligation and puncture(CLP)after 5 days of consecutive administration in anesthetized animals.Twenty mice in each group were randomly selected to observe the survival condition within 7 days after CLP.The bronchoalveolar lavage fluid(BALF)was collected at 24 h after developing the model to determine the concentrations of tumor necrosis factor-alpha(TNF-α),interleukin-1beta(IL-1β),IL-6 and IL-18(by enzyme-linked immunosorbent assay),and the lung tissues were obtained for microscopic examination of the pathological changes which were scored and for determination of wet-to-dry lung weight(W/D)ratio,contents of glutathione(GSH),malondialdehyde(MDA)and iron(by colorimetry),and expression of SIRT1,glutathione peroxidase 4(GPX4)and Acyl-CoA synthetase long chain family member 4(ACSL4)(by Western blot).Results Compared with C group,the 7-day survival rate after CLP was significantly decreased,the concentrations of TNF-α,IL-1β,IL-6 and IL-18 in BALF,W/D ratio and lung injury score were increased,the content of GSH in lung tissues was decreased,the contents of MDA and iron were increased,the expression of SIRT1 and GPX4 was down-regulated,and the expression of ACSL4 was up-regulated in S group(P<0.05).Compared with S group,the 7-day survival rate after CLP was significantly increased,the concentrations of TNF-α,IL-1β,IL-6 and IL-18 in BALF,W/D ratio and lung injury score were decreased,the content of GSH was increased,the contents of MDA and iron were decreased,the expression of SIRT1 and GPX4 was up-regulated,and the expression of ACSL4 was down-regulated in Cur group(P<0.05).Compared with Cur group,the 7-day survival rate after CLP was significantly decreased,the concentrations of TNF-α,IL-1β,IL-6 and IL-18 in BALF,W/D ratio and lung injury score were increased,the content of GSH was decreased,the contents of MDA and iron were increased,the expression of SIRT1 and GPX4 was down-regulated,and the expression of ACSL4 was up-regulated in CE group(P<0.05).Conclusions The mechanism by which curcumin attenuates acute lung injury may be related to activation of SIRT1 and further inhibition of ferroptosis in mice.
作者
陆大浩
黄天丰
张扬
颜克实
高巨
Lu Dahao;Huang Tianfeng;Zhang Yang;Yan Keshi;Gao Ju(Department of Anesthesiology,Northern Jiangsu People's Hospital Affiliated to Yangzhou University,Yangzhou 225001,China;Yangzhou Key Laboratory of Anesthesiology,Yangzhou 225001,China)
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2023年第4期450-454,共5页
Chinese Journal of Anesthesiology
基金
国家自然科学基金(82172190)
江苏省卫生健康委员会医学科研项目面上项目(M2021105)
扬州市重点实验室培育专项基金(YZ20211148)。