文献类型：期刊文献

中文题名：基于蛋白质组学与动物实验探究宣肺化浊丸治疗湿热型肺炎的作用机制

英文题名：Mechanism of XuanFei HuaZhuo Pill for treatment of damp?heat pneumonia based on proteomics and animal experiments

作者：赵佳丽[1,2];张慧娟[1,3];梁夏丽[1,2];王燕如[1,2];田雅文[1,3];张志明[1,4];靳晓杰[1,2,3];刘永琦[1,2]

第一作者：赵佳丽

机构：[1]敦煌医学与转化教育部重点实验室,甘肃中医药大学,兰州730000;[2]甘肃省高校重大疾病分子医学与中医药防治研究重点实验室,兰州730000;[3]甘肃中医药大学药学院,兰州730000;[4]甘肃省中医院,兰州730050

第一机构：甘肃中医药大学科研实验中心（甘肃省中医药标准化技术委员会秘书处）

年份：2026

卷号：34

期号：3

起止页码：382

中文期刊名：中国实验动物学报

外文期刊名：Acta Laboratorium Animalis Scientia Sinica

收录：;北大核心:【北大核心2023】；

基金：甘肃省科技计划项目-科技重大专项(22ZD1FA001);国家中医药管理局科研项目(2021ZYLCYJ08)。

语种：中文

中文关键词：宣肺化浊丸;湿热型肺炎;蛋白质组学;作用机制

外文关键词：XuanFei HuaZhuo Pill;damp?heat pneumonia;proteomics;mechanism

摘要：目的本研究旨在探讨宣肺化浊丸(XuanFei HuaZhuo Pill,XFHZP)对湿热型肺炎(damp-heat pneumonia,DH)作用,并通过蛋白质组学技术揭示其治疗DH的机制,为中医药治疗肺炎提供实验依据。方法建立DH大鼠模型,通过体征观察及评分(一般情况、舌、耳、爪甲、尿液、粪便);肺指数;肺组织苏木素-伊红(HE)染色;肺组织白介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的酶联免疫吸附测定法(ELISA)测定;血细胞计数:白细胞(WBC)、中心粒细胞(Gran)、单核细胞(Mon)、平均血小板体积(MPV);大鼠肺功能检测呼吸频率(F)和分钟通气量(MV)等,明确XFHZP治理DH的疗效。利用Astral DIA定量蛋白组学技术检测空白对照(control,CON)组、DH组和XFHZP+DH组大鼠的右肺下叶,分析CON组和DH组、DH组和XFHZP+DH组之间的差异表达蛋白(DEPs)。并通过生物信息学进行DEPs的KEGG富集分析。采用免疫组化分析代表性DEPs的表达,采用Western Blot对DEPs及其下游蛋白的表达含量进行检测和验证。结果与CON组相比,DH组大鼠出现狂躁、舌色红、耳背血管充盈、尿液色黄及粪便粘滞等症状;肺指数升高,肺组织HE染色出现肺泡壁增厚,炎细胞浸润,肺泡毛细血管充血扩等病变,肺组织中IL-6及TNF-α含量显著升高(P<0.05),WBC、Gran、Mon、MPV及F显著升高(P<0.05),MV显著降低(P<0.05)。与DH组相比,XFHZP+DH组肺指数降低,肺组织HE染色观察到充血情况及肺组织炎性细胞浸润明显减轻,肺组织中IL-6及TNF-α含量显著降低(P<0.05),WBC、Gran、Mon、MPV及F显著降低(P<0.05),MV显著升高(P<0.05)。蛋白质组学分析,表明CON组和DH组大鼠肺组织中有1348个DEPs,DH组和XFHZP+DH组大鼠肺组织中有448个差异蛋白,其中包括AAK1、CACNα1δ1、eIF4E、HSD11β1、ROCK1、TDP1等。DEPs的KEGG通路富集分析显示,cGMP-PKG信号通路和胰岛素信号通路可能是XFHZP治疗DH的关键信号通路。免疫组化结果显示,DH大鼠肺组织中ROCK1和MKNK1的表达均上调,XFHZP治疗后均下调。Western Blot实验进一步表明DH大鼠肺组织中ROCK1和MKNK1的表达均上调,增加了MLC2和eIF4E的磷酸化;XFHZP通过抑制ROCK1和MKNK1的表达,减少MLC2和eIF4E的磷酸化,从而舒张气道平滑肌并抑制炎性因子释放。结论XFHZP对DH具有显著的疗效,其作用机制可能通过调控ROCK1和MKNK1的表达,抑制MLC2和eIF4E的磷酸化,进而舒张气道平滑肌并减轻炎症反应。本研究为XFHZP治疗DH提供了分子层面的科学依据,并为中医药治疗肺炎的机制研究提供了新思路。
Objective To investigate the differential therapeutic effects of XuanFei HuaZhuo pill(XFHZP)on rat pneumonia models with damp?heat pneumonia(DH)syndrome,and to elucidate the underlying mechanisms using proteomics,to provide experimental evidence for the use of traditional Chinese medicine(TCM)in the treatment of pneumonia.Methods A DH group model was established.Therapeutic effects were evaluated via clinical signs and scores(general information,tongue,ear,claw nail,urine,feces),lung index,histopathology(hematoxylin?eosin),enzyme?linked immunosorbent assay for pulmonary interleukin(IL)?6 and tumor necrosis factor(TNF)?α levels,hematology(white blood cell count(WBC),granulocytes(Gran),monocytes(Mon),mean platelet volume(MPV)),and pulmonary function(respiratory frequency(F),minute ventilation(MV)).Astral data?independent acquisition quantitative proteomics was performed on the right inferior lung lobes from control(CON)group,DH group,and XFHZP+DH group to identify differentially expressed proteins(DEPs)between CON vs.DH and DH vs.XFHZP+DH groups.Key DEPs were verified by immunohistochemistry and Western Blot,including their downstream targets(phospho?MLC2,phospho?eIF4E).Results Compared with CON group,DH group exhibited hyperactive behavior,red tongues,ear?vessel dilation,yellow urine,and sticky feces,and displayed elevated lung indices,alveolar wall thickening,inflammatory infiltration,and capillary congestion,plus increased IL?6 and TNF?α(P<0.05),WBC,Gran,Mon,MPV,F(P<0.05),and reduced MV(P<0.05).XFHZP treatment significanly ameliorated lung pathology,reduiced IL?6、TNF?α(P<0.05),and nomalized WBC、Gran、Mon、MPV、F(P<0.05),MV(P<0.05)in DH rats(XFHZP+DH).Proteomics identified 1348 DEPs in the CON vs.DH and 448 in the DH vs.XFHZP+DH group,including AAK1、CACNα2δ1、eIF4E、HSD11β1、ROCK1、TDP1.KEGG analysis highlighted cGMP?dependent protein kinase G and insulin?signaling pathways as potential mechanisms for XFHZP in DH pneumonia.ROCK1/MKNK1 was upregulated in DH lungs,accompanied by increased MLC2/eIF4E phosphorylation,which were suppressed by XFHZP.Conclusions XFHZP exerts significant therapeutic effects on DH syndrome pneumonia,likely by downregulating ROCK1/MKNK1 expression,inhibiting MLC2/eIF4E phosphorylation,relaxing airway smooth muscle,and attenuating inflammation.This study provides molecular?level evidence for XFHZP’s efficacy and offers novel insights into TCM?based pneumonia treatment.