吴茱萸碱通过调节HMGB1/TLR4/NF-κB信号通路减轻大鼠输血相关性急性肺损伤  

Evodiamine alleviates blood transfusion related acute lung injury in Rats by regulating HMGB1/TLR4/NF-κB signaling pathway

作　　者：乔剑 刁春红 于洋 张娜 常梦远 QIAO Jian;DIAO Chunhong;YU Yang(Department of Blood Transfusion Medicine,Joint Support Force 983 Hospital,Tianjin 300142)

机构地区：[1]联勤保障部队第九八三医院输血医学科,天津300142

出　　处：《临床输血与检验》2023年第5期599-607,共9页Journal of Clinical Transfusion and Laboratory Medicine

基　　金：天津市卫生健康委员会科技项目(No.TJWJ2022QN019)资助。

摘　　要：目的 探究吴茱萸碱通过调节高迁移率族蛋白Bl(HMGB1)/Toll样受体4(TLR4)/核转录因子-κB(NF-κB)信号通路减轻大鼠输血相关性急性肺损伤(TRALI)的机制。方法 取72只SD大鼠随机分为对照组、模型组、吴茱萸碱低剂量组、吴茱萸碱高剂量组、空载组、吴茱萸碱高剂量+HMGB1过表达组,每组12只,除对照组外,其余各组大鼠通过腹腔注射脂多糖(LPS)联合输注人血浆的方法制备TRALI模型。分组处理后检测各组大鼠肺功能指标吸气阻力(Ri)、每分钟通气量(MV)、呼气峰流速(PEF);检测各组大鼠肺湿干比,苏木素-伊红(HE)染色检测肺组织病理形态变化;免疫荧光染色检测各组大鼠肺组织内血小板与中性粒细胞聚集情况;检测各组大鼠肺泡灌洗液(BALF)细胞总数和中性粒细胞百分比;酶联免疫吸附测定(ELISA)法检测各组大鼠BALF及血清促炎因子肿瘤坏死因子-α(TNF-α)、白细胞介素1β(IL-1β)水平;免疫印迹检测各组大鼠肺组织HMGB1/TLR4/NF-κB信号通路相关蛋白表达。结果 与模型组相比,吴茱萸碱低剂量组、吴茱萸碱高剂量组大鼠肺功能指标MV、PEF均升高(P <0.05),Ri、肺湿干比、肺组织病理评分、肺组织内血小板-中性粒细胞合光密度、BALF中细胞总数、中性粒细胞百分比、BALF及血清促炎因子TNF-α、IL-1β水平、肺组织HMGB1、TLR4蛋白表达与p-NF-κB p65/NF-κB p65均降低(P <0.05)。HMGB1过表达可减弱吴茱萸碱对TRALI大鼠肺功能的改善作用。结论 吴茱萸碱可能通过抑制HMGB1/TLR4/NF-κB信号激活而抑制TRALI大鼠体内炎症,进而减轻其急性肺损伤,修复其肺功能。Objective To investigate the mechanism by which evodiamine alleviates blood transfusion related acute lung injury(TRALI)in rats by regulating high mobility group protein Bl(HMGB1)/toll like receptor 4(TLR4)/nuclear transcription factor-κB(NF-κB)signaling pathway.Methods A total of 72 SD rats were randomly divided into the model group,low-dose evodiamine group,high-dose evodiamine group,no load group,high-dose evodiamine+HMGB1 overexpression group,and control group with 12 rats in each.Except for the control group,the TRALI model was prepared by intraperitoneal injection of lipopolysaccharide(LPS)combined with infusion of human plasma.After that,we analyzed the lung function indexes such as inspiratory resistance(Ri),minute ventilation(MV)and peak expiratory flow(PEF).Then,in each group we examined the lung wet-dry ratio of rats and the pathological changes of lung tissue by hematoxylin-eosin(HE)staining;the platelet and neutrophil aggregation in lung tissue by immunofluorescence staining;the total number of cells and the percentage of neutrophils in bronchoalveolar lavage fluid(BALF);the levels of pro-inflammatory cytokines tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)in BALF and serum by enzyme-linked immunosorbent assay(ELISA);as well as the expression of HMGB1/TLR4/NF-κB signaling pathway-related proteins in lung tissue by western blotting.Results Compared with the model group,the pulmonary function indexes MV and PEF in low-dose evodiamine group and high-dose evodiamine group were increased(P<0.05),while Ri,lung wet-dry ratio,lung histopathological score,platelet-neutrophil optical density in lung tissue,total number of cells in BALF,percentage of neutrophils,levels of pro-inflammatory factors TNF-αand IL-1βin BALF and serum,HMGB1 and TLR4 protein expression in lung tissue and p-NF-κB p65/NF-κB p65 were all decreased(P<0.05).Overexpression of HMGB1 attenuated the improvement of lung function by evodiamine in TRALI rats.Conclusion Evodiamine may inhibit the inflammation in TRALI rats by in

关 键 词：吴茱萸碱 高迁移率族蛋白Bl/Toll样受体4/核转录因子-κB信号通路 输血相关性急性肺损伤 炎症 肺功能 

分 类 号：R457.13[医药卫生—治疗学]