目的:探讨橘皮素调节硫氧还蛋白互作蛋白(thioredoxin-interacting protein，TXNIP)/含NLR家族PYRIN域蛋白3(NLR family PYRIN domain containing protein 3，NLRP3)信号通路对重症肺炎大鼠细胞焦亡的影响。方法:将SD大鼠分为对照组、重症肺炎组、橘皮素低剂量组、橘皮素高剂量组、橘皮素高剂量+pcDNA组、橘皮素高剂量+pcDNA-TXNIP(TXNIP 激活剂)组，每组 12 只。除对照组外，其它组大鼠均采用气管穿刺肺炎克雷伯菌液的方式构建重症肺炎大鼠模型。造模成功后立即给药处理，给药共持续 4 周。动物肺功能仪监测大鼠呼气峰流速(peak expiratory flow，PEF)、用力肺活量(forced vital capacity，FVC)的变化;ELISA检测大鼠肺泡灌洗液中白细胞介素(interleukin，IL)-1β 和IL-18 水平;检测大鼠肺湿干重比值变化;HE染色检测大鼠肺组织病理变化;碘化丙啶(propidium iodide，PI)染色检测大鼠肺组织中细胞焦亡;Western blot 检测大鼠肺组织中 TXNIP、NLRP3、裂解的天冬氨酸特异性半胱氨酸蛋白酶-1(Cleaved Cysteinyl aspartate-specific proteinase-1，Cleaved Caspase-1)、Gasder-min 家族蛋白D-N端(N-terminal fragment of GSDMD，GSDMD-N)蛋白表达。结果:与对照组比较，重症肺炎组大鼠肺组织严重损伤，PEF、FVC降低，肺泡灌洗液中IL-1β 和 IL-18 水平、肺湿干重比值升高，肺组织中细胞焦亡率及TXNIP、NLRP3、Cleaved Caspase-1、GSDMD-N蛋白表达升高(P＜0。05);与重症肺炎组比较，橘皮素低剂量组、橘皮素高剂量组大鼠肺组织损伤有所改善，PEF、FVC 升高，肺泡灌洗液中IL-1β和IL-18 水平、肺湿干重比值降低，肺组织中细胞焦亡率及 TXNIP、NLRP3、Cleaved Caspase-1、GSDMD-N蛋白表达降低(P＜0。05);pcDNA-TXNIP 逆转了高剂量橘皮素对重症肺炎的保护作用。结论:橘皮素可能通过阻断TXNIP/NLRP3 通路抑制重症肺炎大鼠炎症及细胞焦亡。
Impact of Tangeretin on TXNIP/NLRP3 Signaling Pathway Regulation and Cell Pyroptosis in Severe Pneumonia Rats
Objective:This study aimed to explore the influence of tangeretin on the TXNIP/NLRP3 sig-naling pathway in regulating cell pyroptosis in severe pneumonia rats.Methods:Sprague-Dawley rats were di-vided into control group,severe pneumonia group,low-dose tangeretin group,high-dose tangeretin group,high-dose tangeretin+pcDNA group,and high-dose tangeretin+pcDNA-TXNIP(TXNIP activator)group,with 12 rats in each group.Except for the control group,all other groups were induced to establish a severe pneumonia rat model by tracheal instillation of Klebsiella pneumoniae.Drug administration began immediately after successful modeling and continued for 4 weeks.Lung function parameters including peak expiratory flow(PEF)and forced vital capacity(FVC)were monitored using an animal lung function meter.Levels of inter-leukin(IL)-1β and IL-18 in bronchoalveolar lavage fluid were measured by ELISA.Changes in lung wet-to-dry weight ratio were evaluated.Lung histopathological changes were assessed by HE staining.Cell pyropto-sis in lung tissues was detected using propidium iodide(PI)staining.Expression levels of TXNIP,NLRP3,cleaved caspase-1,and N-terminal fragment of gasdermin D(GSDMD-N)in lung tissues were analyzed by Western blot.Results:Compared with the control group,rats in the severe pneumonia group showed severe lung tissue damage,decreased PEF and FVC,increased levels of IL-1β and IL-18 in bronchoalveolar lavage fluid,increased lung wet-to-dry weight ratio,elevated cell pyroptosis rate in lung tissues,and increased ex-pression of TXNIP,NLRP3,cleaved caspase-1,and GSDMD-N proteins(P＜0.05).Compared with the se-vere pneumonia group,rats in the low-dose tangeretin and high-dose tangeretin groups exhibited improved lung tissue damage,increased PEF and FVC,decreased levels of IL-1β and IL-18 in bronchoalveolar lavage fluid,decreased lung wet-to-dry weight ratio,reduced cell pyroptosis rate in lung tissues,and decreased ex-pression of TXNIP,NLRP3,cleaved caspase-1,and GSDMD-N proteins(P＜0.05).The protective effect of high-dose tangeretin on severe pneumonia was reversed by pcDNA-TXNIP.Conclusion:Tangeretin may inhibit inflammation and cell pyroptosis in severe pneumonia rats by blocking the TXNIP/NLRP3 pathway.
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