肺心汤通过调控NF-κB/NLRP3通路抑制肺动脉平滑肌细胞焦亡而缓解小鼠低氧性肺动脉高压  

Feixin decoction alleviates hypoxic pulmonary hypertension in mice by regulating NF-κB/NLRP3 pathway and inhibiting pyroptosis of pulmo⁃nary artery smooth muscle cells

作　　者：谭骏岚 曹闲雅 郑润锈 易健[1,2] 王飞英 周灵灵 谢思琳 李霞 宋岚 戴爱国 TAN Junlan;CAO Xianya;ZHENG Runxiu;YI Jian;WANG Feiying;ZHOU Lingling;XIE Silin;LI Xia;SONG Lan;DAI Aiguo(Department of Geriatrics,The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410021,China;Hunan Provincial Key Laboratory of Vascular Biology and Translational Medicine,Changsha 410208,China;Department of Respiratory Diseases,Medical School,Hunan University of Chinese Medicine,Changsha 410208,China;Department of Respiratory Medicine,The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410021,China)

机构地区：[1]湖南中医药大学第一附属医院老年病科,湖南长沙410021 [2]血管生物学与转化医学湖南省重点实验室/湖南省高校重点实验室,湖南长沙410208 [3]湖南中医药大学医学院呼吸疾病研究室,湖南长沙410208 [4]湖南中医药大学第一附属医院呼吸科,湖南长沙410021

出　　处：《中国病理生理杂志》2025年第1期36-45,共10页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.82370069);湖南省教育厅优秀青年基金(No.24B0338);湖南省科技攻关项目-芙蓉实验室科技攻关项目(No.2023SK2089)。

摘　　要：目的:基于核因子κB(NF-κB)/核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)信号通路研究肺心汤(FXD)对肺动脉平滑肌细胞(PASMCs)焦亡的调控作用,以探讨FXD缓解小鼠低氧性肺动脉高压(HPH)机制。方法:采用Sugen 5416联合低氧(SuHx)法构建HPH小鼠模型。60只C57BL/6小鼠随机分为对照组、SuHx组、西地那非组,以及低、中、高剂量FXD组,每组10只。给药5周后检测各组小鼠超声心动图指标[包括肺动脉加速时间(PAT)、肺动脉射血时间(PET)、舒张期右心室前壁厚度(RVAWd)和三尖瓣环平面收缩期位移(TAPSE)];右心导管法检测各组小鼠右心室收缩压(RVSP);称量心脏,计算右心肥厚指数(RVHI);HE染色观察肺组织和右心室病理改变;Masson染色检测右心室壁纤维化程度;免疫荧光法检测各组小鼠肺动脉中α-平滑肌肌动蛋白(α-SMA)与NLRP3、消皮素D的N端片段(N-GSDMD)和caspase-1的共定位表达;Western blot法检测肺组织中NF-κB、p-NF-κB、NLRP3、含caspase募集结构域的凋亡相关斑点样蛋白(ASC)、N-GSDMD、白细胞介素1β(IL-1β)、IL-18和cleaved caspase-1蛋白水平;透射电镜观察各组小鼠PASMCs超微形态改变。结果:与对照组相比,SuHx组小鼠RVSP和RVHI升高(P<0.01),右心功能下降(P<0.01),右心室壁纤维化和肺血管重塑增加(P<0.01),肺小动脉α-SMA与NLRP3、N-GSDMD和caspase-1的共定位表达增加(P<0.01),肺组织p-NF-κB、NLRP3、ASC、N-GSDMD、IL-1β、IL-18和cleaved caspase-1蛋白表达增加(P<0.01),并诱导PASMCs焦亡。与SuHx组相比,FXD降低HPH小鼠RVSP和RVHI,改善右心功能,缓解右心室壁纤维化和肺血管血管重塑(P<0.05或P<0.01),减少肺小动脉α-SMA与NLRP3、N-GSDMD和caspase-1的共定位表达(P<0.05或P<0.01),下调肺组织p-NF-κB、NLRP3、ASC、N-GSDMD、IL-1β、IL-18和cleaved caspase-1蛋白表达(P<0.05或P<0.01),并减轻PASMCs焦亡。结论:FXD可通过抑制NF-κB/NLRP3通路减轻PASMCs焦亡,从而缓解SuHx诱导的HPH小鼠肺�AIM:This study aims to investigate the effects of Feixin decoction(FXD)on pyroptosis of pulmonary artery smooth muscle cells(PASMCs)by modulating nuclear factorκB(NF-κB)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)pathway,and to explore how FXD attenuates hypoxic pulmonary hypertension(HPH)in mice.METHODS:A mouse model of HPH was established using the Sugen 5416 combined hypoxia(SuHx)method.Sixty C57BL/6 mice were randomly divided into 6 groups:control group,SuHx group,sildenafil group,and low-,medium-and high-dose FXD groups,with 10 mice in each group.Five weeks after treatment,echocardiographic parameters,including pulmonary artery acceleration time(PAT),pulmonary artery ejection time(PET),right ventricular anterior wall thickness at diastole(RVAWd)and tricuspid annular plane systolic excursion(TAPSE),were measured.Right ventricular systolic pressure(RVSP)was assessed via right heart catheterization.Right ventricular hypertrophy index(RVHI)was determined by weighing the hearts.Histological examination using HE staining was conducted to observe pathological changes in small pulmonary arteries and the right ventricle,while Masson staining was used to assess fibrosis in the right ventricular wall.Immunofluorescence staining was used to detect co-localized expression ofα-smooth muscle actin(α-SMA)with NLRP3,N-terminal fragment of gasdermin D(N-GSDMD)and caspase-1 in the pulmonary arteries.Western blot analysis was conducted to measure the protein levels of NF-κB,p-NF-κB,NLRP3,apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),N-GSDMD,interleukin(IL)-1β,IL-18 and cleaved caspase-1 in lung tissues.Transmission electron microscopy was employed to observe the ultrastructure of PASMCs.RE⁃SULTS:Compared with control group,the mice in SuHx group exhibited elevated RVSP and RVHI(P<0.01),decreased right heart function(P<0.01),increased right ventricular wall fibrosis,and pulmonary vascular remodeling(P<0.01).There was also increased co-localized expressi

关 键 词：肺心汤 低氧性肺动脉高压 肺动脉平滑肌细胞 细胞焦亡 NF-κB/NLRP3通路 

分 类 号：R285.5[医药卫生—中药学] R543.2[医药卫生—中医学] R363