基于空间代谢组学探究川贝母抗肺纤维化的作用机制  

Investigating Mechanism of Fritillariae Cirrhosae Bulbus Against Pulmonary Fibrosis Based on Spatial Metabolomics

作　　者：秦善博 谭鹏 郝露 谢俊杰 林俊芝 张磊[1] 赵军宁[1] QIN Shanbo;TAN Peng;HAO Lu;XIE Junjie;LIN Junzhi;ZHANG Lei;ZHAO Junning(Key Laboratory of Biological Evaluation of Traditional Chinese Medicine(TCM)Quality of National Administration of TCM,Sichuan Academy of Chinese Medicine Sciences,Chengdu 610041,China;College of Food and Biological Engineering,Chengdu University,Chengdu 610106,China;TCM Regulating Metabolic Diseases Key Laboratory of Sichuan Province,Hospital of Chengdu University of TCM,Chengdu 610075,China)

机构地区：[1]四川省中医药科学院国家中医药管理局中药质量生物评价重点研究室,成都610041 [2]成都大学食品与生物工程学院,成都610106 [3]成都中医药大学附属医院,代谢性疾病中医药调控四川省重点实验室,成都610075

出　　处：《中国实验方剂学杂志》2024年第13期150-159,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：四川省科技厅重点研发项目(2022YFS0429);中央引导地方科技发展项目(2022ZYD0102);四川省中医药管理局项目(2023MS496);四川省科研院所基本科研项目(2022JDKY0036)。

摘　　要：目的:基于空间代谢组学技术结合药理指标,解析川贝母粉改善博莱霉素诱导的大鼠肺纤维化的作用机制。方法:将60只SD大鼠随机分为空白组、模型组及川贝母高、中、低剂量组,每组12只。除空白组外,其他组大鼠均采用气管注射博莱霉素建立肺纤维化模型。术后川贝母高、中、低剂量组分别给予0.36、0.18、0.09 g·kg^(-1)川贝母粉的水溶液,连续给药28 d,空白组和模型组灌胃给予等量蒸馏水。末次给药后收集肺组织及血液,通过苏木素-伊红(HE)、马松(Masson)染色综合评估大鼠肺组织病理状况,利用空气动力辅助解吸电喷雾离子化质谱成像技术对不同实验组大鼠肺组织进行质谱成像分析,依据HE染色图选择模型组和川贝母高剂量组肺组织纤维化区域进行空间代谢组学分析,采用有监督的正交偏最小二乘法-判别分析(OPLS-DA)获得的变量重要性投影(VIP)值>1、t检验P<0.05及变异倍数分析来筛选组间差异代谢产物;采用京都基因与基因组百科全书(KEGG)数据库对筛选出的差异代谢物进行代谢通路分析。采用蛋白免疫印迹法(Western blot)检测大鼠肺组织中核转录因子-κB p65(NF-κB p65)和血红素加氧酶-1(HO-1)蛋白表达;生化检测大鼠肺组织中超氧化物歧化酶(SOD)、丙二醛(MDA)和谷胱甘肽(GSH)水平;采用酶联免疫吸附测定法(ELISA)检测大鼠血清中白细胞介素(IL)-1β、IL-6、核因子E2相关因子2(Nrf2)、肿瘤坏死因子-α(TNF-α)水平,对筛选出的部分相关性强的信号通路进行验证。结果:质谱成像实验结果显示,肺纤维化大鼠在灌胃川贝母粉28 d后,其肺组织纤维化区域的L-精氨酸含量与模型组大鼠比较具有明显差异,磷脂酰胆碱含量低于模型组大鼠肺纤维化区域中磷脂酰胆碱含量。Western blot结果表明,与模型组比较,在灌胃给药川贝母粉28 d后,可以抑制肺纤维化大鼠肺组织中NF-κB p65蛋白表达量上升,高�Objective:Based on spatial metabolomics technology combined with pharmacological indexes,to analyze the mechanism of Fritillariae Cirrhosae Bulbus(FCB)powder in improving bleomycininduced pulmonary fibrosis in rats.Method:Sixty SD rats were randomly divided into five groups,including the blank group,the model group,and high,medium,low dosage groups of FCB.Except for the blank group,rats in all other groups were injected with bleomycin by tracheal injection to establish a pulmonary fibrosis model.Postoperatively,the high,medium and low dosage groups of FCB were administered aqueous solutions of FCB powder at doses of 0.36,0.18,0.09 g·kg^(-1),respectively,continuously for 28 d.The blank and model groups were given an equal volume of distilled water by gavage.After the last administration,lung tissues and blood samples were collected,the pathological conditions of rat lung tissues were comprehensively evaluated by hematoxylin-eosin(HE)and Masson staining,and aerodynamic assisted desorption electrospray ionization mass spectrometry imaging(AFADESI-MSI)was used for MSI of rat lung tissues from different experimental groups.Spatial metabolomics analysis was conducted on the fibrotic areas of lung tissues in the model group and the high dosage group of FCB based on HE staining images.Differential metabolites between groups were screened by orthogonal partial least squares-discriminant analysis(OPLS-DA),with variable importance in the projection(VIP)values>1,t-test P<0.05,and fold change analysis.Metabolic pathway analysis of the identified differential metabolites was performed using Kyoto Encyclopedia of Genes and Genomes(KEGG).Protein expression levels of nuclear transcription factor-κB p65(NF-κB p65)and heme oxygenase-1(HO-1)in rat lung tissues were detected by Western blot.Biochemical assessments of superoxide dismutase(SOD),malondialdehyde(MDA)and glutathione(GSH)levels in rat lung tissues were conducted.Serum levels of interleukin(IL)-1β,IL-6,nuclear factor erythroid 2 related factor 2(Nrf2),and tumor necrosis

关 键 词：川贝母 肺纤维化 空间代谢组学 炎症因子 氧化应激 质谱成像 博莱霉素 

分 类 号：R22[医药卫生—中医基础理论] R28[医药卫生—中医学] R563[理学—分析化学] O657[理学—化学]