摘要
目的探讨毛蕊花苷(AC)对肺炎链球菌(SP)肺炎大鼠肺损伤及肌醇需求酶1α/硫氧还蛋白结合蛋白/核苷酸结合寡聚化结构域样受体蛋白3(IRE1α/TXNIP/NLRP3)信号通路的影响。方法构建SP肺炎模型大鼠,将造模成功大鼠随机分为模型组(Model组)、毛蕊花苷低、高剂量组(AC-L、AC-H组)、毛蕊花苷高剂量+空载体组(AC-H+pAdEasyTM组)、毛蕊花苷高剂量+IRE1α过表达腺病毒载体组(AC-H+Ad-IRE1α组),每组18只,另取18只健康正常大鼠作为对照组(Control组);检测各组大鼠氧合指数;酶联免疫吸附试验(ELISA)检测肺泡灌洗液炎症因子水平;检测各组大鼠肺组织湿/干比(W/D)及氧化应激水平;苏木精-伊红(HE)染色观察肺组织病理情况;原位缺口末端标记法(TUNEL)染色检测肺组织细胞凋亡;蛋白免疫印迹法(Western blot)检测IRE1α/TXNIP/NLRP3信号通路相关蛋白表达检测。结果Model组较Control组肺组织损伤明显,肺泡上皮细胞肿胀,排列杂乱,肺泡壁毛细血管扩张,肺泡间隔增厚,炎性细胞浸润明显,PaCO_(2)、TNFα、IL-1β、IL-6、W/D、MDA水平和细胞凋亡率及Bax、p-IRE1α/IRE1α、TXNIP、NLRP3表达升高,PaO_(2)、OI、SOD水平及Bcl-2表达降低(P<0.05);AC-L、AC-H组较Model组肺组织形态有所改善,肺泡间隔少量增厚,少量炎性细胞浸润,PaCO_(2)、TNFα、IL-1β、IL-6、W/D、MDA水平和细胞凋亡率及Bax、p-IRE1α/IRE1α、TXNIP、NLRP3表达降低,PaO_(2)、OI、SOD水平及Bcl-2表达升高(P<0.05);AC-H+Ad-IRE1α组较AC-H+pAdEasyTM组PaCO_(2)、TNFα、IL-1β、IL-6、W/D、MDA水平和细胞凋亡率及Bax、p-IRE1α/IRE1α、TXNIP、NLRP3表达升高,PaO_(2)、OI、SOD水平及Bcl-2表达降低(P<0.05)。结论毛蕊花苷可改善SP肺炎大鼠肺损伤,且证明与抑制IRE1α/TXNIP/NLRP3信号通路相关。
OBJECTIVE To investigate the effect of verbascoside(AC)on lung injury and inositol requiring enzyme 1α/thioredoxin-interacting protein/nucleotide binding oligomerization domain-like receptor protein 3(IRE1α/TXNIP/NLRP3)signaling pathway in Streptococcus pneumoniae(SP)pneumonia rats.METHODS SP pneumonia model rats were constructed,and successfully modeled rats were randomly assigned into Model group,low and high dose verbascoside groups(AC-L,AC-H groups),high dose verbascoside+empty vector group(AC-H+pAdEEasyTM group),and high dose verbascoside+IRE1αoverexpression adenovirus vector group(AC-H+Ad-IRE1αgroup),with 18 rats in each group.Another 18 healthy normal rats were as the Control group.The oxygenation index of rats in each group was measured.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of inflammatory factors in bronchoalveolar lavage fluid.The wet/dry ratio(W/D)and oxidative stress level of lung tissue of rats in each group were detected.Hematoxyl-eosin(HE)staining was used to observe the pathological condition of lung tissue.Tdt mediated dutp nick end labeling(TUNEL)staining was used to detect apoptosis in lung tissue cells.Western blot was used to detect the expression of proteins related to the IRE1α/TXNIP/NLRP3 signaling pathway.RESULTS The lung tissue injury in the Model group was more significant than that in the Control group,with swollen and disordered alveolar epithelial cells,dilated alveolar wall capillaries,thickened alveolar septa,and significant infiltration of inflammatory cells,the PaCO_(2),levels of TNFα,IL-1β,IL-6,W/D,MDA,apoptosis rate,and the expression of Bax,p-IRE1α/IRE1α,TXNIP,and NLRP3 were higher,and the PaO_(2),OI,level of SOD,and the expression of Bcl-2 were lower(P<0.05).The lung tissue morphology of the AC-L and AC-H groups improved compared with the Model group,with slight thickening of the alveolar septa and infiltration of inflammatory cells,the PaCO_(2),levels of TNFα,IL-1β,IL-6,W/D,MDA,apoptosis rate,and the expression of Bax,p-IRE1α/IRE1α,TXNIP,and NLRP3 were lower,and the PaO_(2),OI,level of SOD,and the expression of Bcl-2 were higher(P<0.05).The PaCO_(2),levels of TNFα,IL-1β,IL-6,W/D,MDA,apoptosis rate,and the expression of Bax,p-IRE1α/IRE1α,TXNIP,and NLRP3 in the AC-H+Ad-IRE1αgroup were higher than those in AC-H+pAdEasyTM group,and the PaO_(2),OI,level of SOD,and the expression of Bcl-2 were lower(P<0.05).CONCLUSION Verbascoside can improve lung injury in SP pneumonia rats,and its related to the inhibition of the IRE1α/TXNIP/NLRP3 signaling pathway.
作者
陈丽媛
车姣子
朱翠敏
CHEN Liyuan;CHE Jiaozi;ZHU Cuimin(Department of Clinical Laboratory,Chengde Central Hospital,Chengde 067000,China;Department of Oncology,Affiliated Hospital of Chengde Medical College,Chengde 067000,China)
出处
《中国药学杂志》
北大核心
2025年第18期1931-1937,共7页
Chinese Pharmaceutical Journal
基金
河北省医学科学研究重点课题计划项目资助(编号:20211664)。
关键词
毛蕊花苷
肌醇需求酶1α/硫氧还蛋白结合蛋白/核苷酸结合寡聚化结构域样受体蛋白3信号通路
肺炎链球菌
肺炎
肺损伤
verbascoside
inositol requiring enzyme 1α
thioredoxin-interacting protein
nucleotide binding oligomerization domain-like receptor protein 3 signaling pathway
Streptococcus pneumoniae
pneumonia
lung injury
