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田慧,陈光宇,瞿昊宇,何群,谢梦洲.UPLC-Q-TOF-MS结合网络药理学及实验验证探讨益气平喘豆乳粉药物成分及其干预COPD的作用机制[J].湖南中医药大学学报英文版,2023,43(11):2092-2103.[Click to copy
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| UPLC-Q-TOF-MS结合网络药理学及实验验证探讨益气平喘豆乳粉药物成分及其干预COPD的作用机制 |
| 田慧,陈光宇,瞿昊宇,何群,谢梦洲 |
| (湖南中医药大学, 湖南 长沙 410208;湖南省药食同源功能性食品工程技术研究中心, 湖南 长沙 410208;湖南中医药大学中医心肺病证辨证与药膳食疗重点研究室, 湖南 长沙 410208) |
| 摘要: |
| 目的 运用UPLC-Q-TOF-MS、网络药理学和实验验证探讨益气平喘豆乳粉干预慢性阻塞性肺疾病(chronic obstructive pulmonary disease, COPD)的作用机制。方法 采用UPLC-Q-TOF-MS解析益气平喘豆乳粉中药复方中的活性成分,在SwissTargetPrediction中筛选靶点;在GeneCards中收集COPD靶点,运用Venny 2.1.0平台,对活性成分靶点与COPD靶点取交集获得潜在靶点;将潜在靶点导入STRING和DAVID中进行蛋白质相互作用网络分析(protein-protein interaction, PPI)、GO和KEGG分析。将36只小鼠按随机数字表法分为正常组、模型组、高剂量组(12.0 mL/kg)、中剂量组(6.0 mL/kg)、低剂量组(3.0 mL/kg)、地塞米松组(1.0 mg/kg),每组6只。1天1次,连续给药30 d。除正常组外,其余小鼠采用脂多糖(lipopolysaccharide, LPS)联合香烟烟雾诱导法建立COPD小鼠模型。造模成功后,每周测量1次小鼠体质量,末次给药后,HE染色观察肺组织及脂肪组织形态变化;ELISA检测血清及支气管肺泡(bronchoalveolar lavage fluid, BALF)灌洗液中炎症因子TNF-α、IL-6水平;RT-PCR检测肺组织中TNF-α、ICAM-1、SELE mRNA水平。结果 UPLC-Q-TOF-MS分析得到143个活性成分,297个潜在交集靶点。PPI、GO和KEGG分析发现,益气平喘豆乳粉可能通过作用于TNF、AKT1、IL-6等靶点,调节TNF、PI3K-Akt、癌症等信号通路以干预COPD病理机制。动物实验结果显示,与正常组相比,模型组小鼠肺组织可见大量炎性细胞浸润,脂肪组织中空泡脂滴增多,体质量明显降低(P＜0.01),血清中TNF-α和IL-6含量升高(P＜0.01)。与模型组相比,各用药组肺组织炎症细胞浸润程度减轻,脂肪细胞分布增多排列紧密,中剂量组小鼠体质量降低最少(P＜0.05),高剂量组小鼠血清中TNF-α含量显著降低(P＜0.05),各用药组肺组织中ICAM-1 mRNA的相对表达量显著降低(P＜0.01)。结论 益气平喘豆乳粉可能通过干预TNF、PI3K-Akt通路,抑制TNF-α、IL-6的释放,发挥抗炎、改善营养不良等作用。 |
| 关键词: UPLC-Q-TOF-MS 网络药理学 益气平喘豆乳粉 COPD小鼠模型 营养不良 药食同源 |
| DOI：10.3969/j.issn.1674-070X.2023.11.023 |
| Received:April 17, 2023 |
| 基金项目:2021年湖南省研究生科研创新项目（CX20210706）。 |
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| Pharmacological ingredients of Yiqi Pingchuan aSoybean Milk Powder and its intervention mechanism on COPD based on UPLC-Q-TOF-MS combined with network pharmacology and experimental validation |
| TIAN Hui,CHEN Guangyu,QU Haoyu,HE Qun,XIE Mengzhou |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Hunan Engineering Research Center of Medicine-Food Homologous Functional Food, Changsha, Hunan 410208, China;Key Laboratory of TCM Heart and Lung Pattern Differentiation, Medicated Diet & Dietotherapy, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To explore the intervention mechanism of Yiqi Pingchuan Soybean Milk Powder (YQPCSMP) on chronic obstructive pulmonary disease (COPD) based on UPLC-Q-TOF-MS, network pharmacology, and experimental validation. Methods UPLC-Q-TOF-MS was used to analyze the active ingredients in TCM formula of YQPCSMP and screen the targets in SwissTargetPrediction. Venny 2.1.0 platform was used to intersect active ingredient targets and COPD targets collected in GeneCards to obtain potential targets, which were introduced into STRING and DAVID for Protein-Protein Interaction Networks (PPI), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. A total of 36 mice were assigned into normal, model, high-dose (12.0 mL/kg), medium-dose (6.0 mL/kg), low-dose (3.0 mL/kg), and dexamethasone groups (1.0 mg/kg) by random number table method, with 6 mice in each group. The drug was administered once a day for 30 days. Except for the normal group, the other mice were applied with the "Lipopolysaccharide (LPS) and cigarette smoke induction method" to establish a COPD mouse model. The body mass of mice was measured once a week after successful modeling. The morphological changes of lung and adipose tissues were observed by HE staining after the last dose. Combined with the network pharmacology results obtained from previous studies, inflammatory factors of TNF-α and IL-6 in serum and bronchoalveolar lavage fluid (BALF) were checked by ELISA, and the mRNA levels of TNF-α, ICAM-1, and SELE in lung tissue were tested by RT-PCR. Results A total of 143 active ingredients and 297 potential intersection targets were obtained by UPLC-Q-TOF-MS. PPI, GO, and KEGG analyses indicated that YQPCSMP might intervene in the pathological mechanism of COPD by acting on targets such as TNF, AKT1, and IL6 and regulating signal pathways such as TNF, PI3K-Akt, and cancer. The results of animal experiments showed that compared with the normal group, the mice in the model group showed a large number of inflammatory cells infiltration in the lung tissue, an increase in hollow vesicle lipid droplets in adipose tissue, a significant decrease in body mass (P＜0.01), and an increase of TNF-αand IL-6 content (P＜0.01) in serum. Compared with model group, each treatment group showed a reduction in the infiltration degree of inflammatory cells in the lung tissue as well as a rising distribution and tight arrangement of adipocytes. The body mass of the medium-dose group decreased the least (P＜0.05), while the content of TNF-α in serum of the high-dose group mice significantly was lower (P＜0.05). The relative expression of ICAM-1 mRNA in lung tissues of all treatment groups was significantly lower (P＜0.01). Conclusion YQPCSMP might inhibit the release of TNF-α and IL-6 by intervening TNF and PI3K-Akt pathways, and thus exert anti-inflammatory and malnutrition-improving effects. |
| Key words: UPLC-Q-TOF-MS network pharmacology Yiqi Pingchuan Soybean Milk Powder COPD mouse model malnutrition homology of medicine and food |
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