右美托咪定通过调控TXNIP/NLRP3炎症小体途径减轻脂多糖诱导的小鼠肠道屏障损伤  

Dexmedetomidine reduces lipopolysaccharide-induced intestinal barrier damage in mice by regulating TXNIP/NLRP3 inflammasome pathway

作　　者：贾彤[1] 邢珍[1] 王会娟[2] 李国利[1] JIA Tong;XING Zhen;WANG Hui-juan;LI Guo-li(Department of Anesthesiology,The First Affiliated Hospital of Hebei North University,Zhangjiakou 075061,China;Department of Critical Care Medicine,The First Affiliated Hospital of Hebei North University,Zhangjiakou 075061,China)

机构地区：[1]河北北方学院附属第一医院麻醉科,河北张家口075061 [2]河北北方学院附属第一医院重症医学科,河北张家口075061

出　　处：《中国病理生理杂志》2021年第9期1589-1595,共7页Chinese Journal of Pathophysiology

基　　金：张家口市科技计划项目(No.1621066D);河北省科技计划项目(No.162777173);2020年河北省政府资助临床医学人才培养项目(冀卫办科教〔2021〕1号)。

摘　　要：目的:探讨右美托咪定(DEX)调控硫氧还蛋白相互作用蛋白(TXNIP)/核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体途径减轻脂多糖(LPS)诱导的小鼠肠道屏障损伤的机制。方法:实验小鼠分为对照组、模型组、DEX组、DEX+pcDNA3.1组和DEX+TXNIP组,每组12只。除对照组外,其余各组小鼠腹腔注射15 mg/kg LPS,对照组小鼠腹腔注射等体积生理盐水。在LPS诱导肠道屏障损伤前30 min,DEX组小鼠腹腔注射30μg/kg DEX,DEX+pcDNA3.1组和DEX+TXNIP组在DEX组基础上分别皮下注射5 nmol pcDNA3.1空载体及5 nmol pcDNA3.1-TXNIP过表达载体,对照组和模型组小鼠则皮下注射等体积生理盐水。收集小鼠血清及肠道组织样本,采用ELISA检测血清中白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)的水平,苏木精-伊红(HE)染色观察肠道组织形态,采用相应试剂盒检测肠道组织中氧化应激指标丙二醛(MDA)、活性氧(ROS)、超氧化物歧化酶(SOD)和谷胱甘肽(GSH)水平,采用RT-qPCR和Western blot分别检测肠道组织中TXNIP、NLRP3、caspase-1和IL-1β的mRNA和蛋白水平。结果:模型组肠道组织完整性被破坏,出现明显的细胞脱落现象,炎症浸润明显,病理评分升高(P<0.05);DEX组和DEX+pcDNA3.1组肠道组织完整,但出现明显的炎症浸润现象,与模型组相比病理评分降低(P<0.05);DEX+TXNIP组肠道完整性被破坏,出现细胞脱落现象,炎症浸润明显,与DEX+pcDNA3.1组相比病理评分升高(P<0.05)。与对照组相比,模型组血清中IL-1β和TNF-α水平升高,肠道组织中MDA含量、ROS水平及TXNIP、NLRP3、caspase-1和IL-1β的mRNA和蛋白水平升高(P<0.05),肠道组织中SOD和GSH活性降低(P<0.05);与模型组相比,DEX组和DEX+pcDNA3.1组血清中IL-1β和TNF-α水平降低,肠道组织中MDA含量、ROS水平及TXNIP、NLRP3、caspase-1和IL-1β的mRNA和蛋白水平降低(P<0.05),肠道组织中SOD和GSH活性升高(P<0.05);分别与DEX组和DEX+pcDNA3.1组相比,DEX+TXNIP组血�AIM:To investigate the mechanism of dexmedetomidine(DEX)reducing the intestinal barrier damage induced by lipopolysaccharide(LPS)in mice by regulating the thioredoxin-interacting protein(TXNIP)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome pathway.METHODS:The mice were divided into control group,model group,DEX group,DEX+pcDNA3.1 group and DEX+TXNIP group,with 12 mice in each group.Except for control group,the mice in other groups were intraperitoneally injected with LPS at 15 mg/kg,and the mice in control group were injected with equal volume of normal saline.At 30 min before LPS-induced intestinal barrier injury,the mice in DEX group were intraperitoneally injected with 30μg/kg DEX(30 mg/L),the mice in DEX+pcDNA3.1 group and DEX+TXNIP group were subcutaneously injected with 5 nmol pcDNA3.1 empty vector and pcDNA3.1-TXNIP overexpression vector on the basis of DEX group,respectively,and the mice in control group and model group were injected with equal volume of saline.The serum levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)were measured by ELISA.Hematoxylin-eosin(HE)staining was used to observe the morphological changes of intestinal tissues.The levels of oxidative stress indicators,malondialdehyde(MDA),reactive oxygen species(ROS),superoxide dismutase(SOD)and glutathione(GSH),in intestinal tissues were detected using respective kits.The mRNA and protein levels of TXNIP,NLRP3,caspase-1 and IL-1β in intestinal tissues were detected by RT-qPCR and Western blot,respectively.RESULTS:The integrity of the intestinal tissue in model group was destroyed,with obvious cell shedding and inflammatory infiltration,and the pathological score was increased(P<0.05).Compared with model group,the intestinal tissues in DEX group and DEX+pcDNA3.1 group were intact,but there was obvious inflammatory infiltration,and the pathological score was decreased(P<0.05).In DEX+TXNIP group,the integrity of the intestine was destroyed,cell shedding and inflammatory infiltration wer

关 键 词：右美托咪定 硫氧还蛋白相互作用蛋白 NLRP3炎症小体 肠道屏障损伤 脂多糖 氧化应激 

分 类 号：R574.4[医药卫生—消化系统] R363.2[医药卫生—内科学]