摘要
目的从“肺-肠轴”角度探讨玉屏风颗粒防治慢性阻塞性肺疾病(COPD)的可能作用机制。方法32只大鼠随机分为正常组、模型组、罗红霉素组、玉屏风颗粒组,每组8只。除正常组外,其余各组大鼠采用气管滴注脂多糖(LPS)联合烟熏法制备COPD大鼠模型,持续12周。造模第5~12周,罗红霉素组和玉屏风颗粒组分别给予相应药物31.5 mg/(kg·d)、1.575 g/(kg·d)灌胃,正常组和模型组给予10 ml/(kg·d)生理盐水。末次给药后24 h取材。HE染色观察各组大鼠肺组织病理形态改变情况。正常组、模型组及玉屏风颗粒组大鼠应用超高效液相色谱-四极杆飞行时间质谱联用技术(UHPLC-QTOFMS)检测肺泡灌洗液(BALF)差异性代谢物;16S rDNA测序技术检测肠道菌群的差异菌属,并将差异性代谢物和差异菌属进行关联分析。结果与正常组比较,模型组大鼠小支气管壁杯状细胞增多,支气管壁平滑肌层消失,可见炎性细胞浸润;与模型组比较,罗红霉素组肺泡间质轻微水肿,炎性细胞明显减少;玉屏风颗粒组未见明显肺泡间质水肿,伴有少量炎性细胞浸润。BALF差异性代谢物筛选结果显示,与正常组比较,模型组中12种物质上调,19种物质下调;与模型组比较,玉屏风颗粒组中37种物质上调,43种物质下调。KEGG分析共得出2条代谢通路,分别是甘油磷脂代谢、不饱和脂肪酸的生物合成代谢;与模型组比较,玉屏风颗粒组胆碱、乙酰胆碱、甘油-3-磷酸、甘油磷酸胆碱、棕榈酸、花生四烯酸呈上升趋势;硬脂酸、二十二碳六烯酸呈下降趋势(P<0.05)。肠道菌群结果显示,正常组和模型组有80种差异物种,模型组和玉屏风颗粒组有65种差异物种;与模型组比较,玉屏风颗粒组相对丰度水平前五位的物种中普雷沃菌属_9、Ruminococcaceae_UCG-005、Ruminiclostridium_6水平升高,乳杆菌属、拟杆菌属水平降低(P<0.05)。相关性分析结果显示,模型组vs正常组,花生四烯酸与Oribacterium呈负相关(r=-0.753,P<0.01);玉屏风颗粒组vs模型组,硬脂酸与拟杆菌属(r=0.788)、分枝杆菌属(r=0.826)、[Eubacterium]_ruminantium_group(r=0.770)均呈正相关(P<0.01);花生酸与Roseiarcus(r=-0.779)、甘油-3-磷酸与脱硫弧菌属(r=-0.758)、花生四烯酸与Oribacterium(r=-0.753)、棕榈酸与Pseudolabrys(r=-0.750)均呈负相关(P<0.01)。结论玉屏风颗粒可影响COPD模型大鼠BALF代谢产物和肠道微生物的菌群结构,调节“肺-肠轴”平衡可能是其防治COPD的可能作用机制之一。
Objective To explore the possible mechanism of Yupingfeng Granules(玉屏风散)in preventing and treating chronic obstructive pulmonary disease(COPD)from the perspective of“lung-gut axis”.Methods Thirty-two male Wistar rats were randomly divided into normal group,model group,roxithromycin group and Yupingfeng Granules group,with 8 rats in each group.Except for the normal group,the rat model of COPD was prepared by intra⁃tracheal instillation of lipopolysaccharide(LPS)combined with smoking for 12 weeks.Since the fifth week of model⁃ing,the roxithromycin group and the Yupingfeng Granules group were given 31.5 mg/(kg·d)and 1.575 g/(kg·d)of corresponding drugs respectively by gavage,and normal group and model group were given 10 ml/(kg·d)physiolo-gical saline.Sample was collected 24 hours after the last administration.The pathological changes of lung tissue were observed using HE staining;Ultrahigh performance liquid chromatography quadrupole time of flight mass spectrome⁃try(UHPLC-QTOFMS)was used to detect the differential metabolites in alveolar lavage fluid(BALF)in all groups but roxithromycin group;16S rDNA sequencing technology was used to detect the changes of intestinal flora,and the association analysis was conducted between the differential metabolites and the differential flora.Results Compared with the normal group,the model group showed an increase in goblet cells in the small bronchial wall,disappearance of the smooth muscle layer of the bronchial wall,and infiltration of inflammatory cells;compared with the model group,roxithromycin group showed slight alveolar interstital edema,and obviously reduced inflammatory cell,while no obvious alveolar interstital edema was observed in the Yupingfeng Granules group,showing a small amout of inflam⁃matory cell infiltration.The results of the BALF differential metabolite screening showed that compared with the nor⁃mal group,12 substances were upregulated and 19 substances were downregulated in the model group;compared with the model group,37 substances in the Yupingfeng Granules group were upregulated and 43 substances were downregu⁃lated KEGG analysis yielded a total of 2 metabolic pathways,glycerophospholipid metabolism,and unsaturated fatty acid biosynthetic metabolism;compared with the model group,choline,acetylcholine,glycerol-3-phosphate,glycerophosphate choline,palmitic acid,and arachidonic acid showed an upward trend,while stearic acid and doco⁃sahexaenoic acid showed a downward trend in Yupingfeng Granules group(P<0.05).The results of the intestinal flora showed that,there are 80 different species between the normal group and the model group,and 65 different species between the model group and Yupingfeng Granules group.Among the top 5 species with relative abundance levels,compared with the model group,the level of Prevotella_9,Ruminococcaceae_UCG-005,Ruminiclostridium_6 increase,and Lactobacillus,Bacteroides decrease(P<0.05).The results of the correlation analysis showed that,in the normal and model groups,arachidonic acid was negatively correlated with Oribacterium(r=-0.753,P<0.01);in the Yupingfeng Granules group and model group,stearic acid and Bacteroides(r=0.788),Mycobacterium(r=0.826),[Eubacterium]_Ruminantium_Group(r=0.770)was positively correlated(P<0.01),Arachidic acid was negatively correlated with Roseiarcus(r=-0.779),glycerol-3-phosphate was negatively correlated with Desulfovibrio(r=-0.758),Arachidonic acid was negatively correlated with Oribacterium(r=-0.753),and Palmitic acid was negatively correlated with Pseudolabs(r=-0.750,P<0.01).Conclusion Yupingfeng Granules can affect the metabolism of BALF and the flora structure of intestinal microorganisms,and regulating the balance of“lung-gut axis”may be one of the mechanisms of Yupingfeng Granules in treatment of COPD.
作者
刘海叶
骆姗
贾智玲
董文军
王媛媛
王杰鹏
于文涛
LIU Haiye;LUO shan;JIA Zhiling;DONG Wenjun;WANG Yuanyuan;WANG Jiepeng;YU Wentao(Hebei College of Traditional Chinese Medicine,Shijiazhuang,050200)
出处
《中医杂志》
CSCD
北大核心
2023年第20期2116-2124,共9页
Journal of Traditional Chinese Medicine
基金
河北省中医药管理局科研计划(2019072)
河北中医学院博士科研基金(BSZ2018013)。
关键词
慢性阻塞性肺疾病
玉屏风颗粒
肺-肠轴
代谢组学
肠道菌群
chronic obstructive pulmonary disease
Yupingfeng Granules
lung-gut axis
metabonomics
gut micro⁃flora
作者简介
通讯作者:于文涛,ywtawen@163.com。
