Hong Du , Meijuan Shao , Shangcheng Xu , Qian Yang , Jingping Xu , Hong Ke , Li Zou , Liping Huang , Yanru Cui , Fei Qu
{"title":"结合代谢组学和网络药理学分析探索葛根抗肺纤维化的机制:精氨酸代谢途径的参与","authors":"Hong Du , Meijuan Shao , Shangcheng Xu , Qian Yang , Jingping Xu , Hong Ke , Li Zou , Liping Huang , Yanru Cui , Fei Qu","doi":"10.1016/j.jep.2024.118346","DOIUrl":null,"url":null,"abstract":"<div><h3>Ethnopharmacological relevance</h3><p>Pueraria lobata (Willd.) Ohwi is a typical medicinal and edible plant with a long application history in China and Southeast Asia. As a widely used traditional medicine, <em>P. lobata</em> exhibits the properties of anti-inflammatory, antipyretic, antioxidant, relieving cough and asthma. Particularly, the increasing evidence indicates that the <em>P. lobata</em> has the therapeutic effect on fibrotic-related diseases in terms of metabolic regulation. However, the mechanisms of <em>P. lobata</em> on pulmonary fibrosis (PF) has not been thoroughly explored.</p></div><div><h3>Aim of the study</h3><p>This study aimed to explore the effect of arginine metabolites of <em>P. lobata</em> against PF model by integrating metabolomics and network pharmacology analysis. It might provide a new idea for the target finding of <em>P. lobata</em> anti-pulmonary fibrosis.</p></div><div><h3>Materials and methods</h3><p>In this study, the Sprague Dawley (SD) rats were randomly divided into five experimental groups: saline-treated control group, bleomycin-induced fibrosis group, prednisolone acetate group, <em>P. lobata</em> 3.2 g/kg group and <em>P. lobata</em> 6.4 g/kg group. The therapeutic effect of <em>P. lobata</em> on bleomycin-induced PF in rats was evaluated by clinical symptoms such as lung function, body weight, hematoxylin eosin staining (HE), Masson staining and hydroxyproline assay. Next, the plasma metabolomics analysis was carried out by LC-MS to explore the pathological differences between the group of control, PF and <em>P. lobata</em>-treated rats. Then, the network pharmacology study coupled with experimental validation was conducted to analysis the results of metabolic research. We constructed the “component-target-disease” network of <em>P. lobata</em> in the treatment of PF. In addition, the molecular docking method was used to verify the interaction between potential active ingredients and core targets of <em>P. lobata</em>. Finally, we tested NOS2 and L-OT in arginine-related metabolic pathway in plasma of the rats by enzyme-linked immunosorbent assay (ELISA). Real-time PCR was performed to observe the level of TNF-α mRNA and MMP9 mRNA. And we tested the expression of TNF-α and MMP9 by Western blot analysis.</p></div><div><h3>Results</h3><p>Our findings revealed that <em>P. lobata</em> improved lung function and ameliorated the pathological symptoms, such as pathological damage, collagen deposition, and body weight loss in PF rats. Otherwise, the plasma metabolomics were employed to screen the differential metabolites of amino acids, lipids, flavonoids, arachidonic acid metabolites, glycoside, etc. Finally, we found that the arginine metabolism signaling mainly involved in the regulating of <em>P. lobata</em> on the treatment of PF rats. Furtherly, the network pharmacology predicted that the arginine metabolism pathway was contained in the top 20 pathways. Next, we integrated metabolomics and network pharmacology that identified NOS2, MMP9 and TNF-α as the <em>P. lobata</em> regulated hub genes by molecular docking. Importantly, it indicated a strong affinity between the puerarin and the NOS2. <em>P. lobata</em> attenuated TNF-α, MMP-9 and NOS2 levels, suppressed TNF-α and MMP-9 protein expression, and decreased L-OT and NOS2 content in PF rats. These results indicated that the effects of <em>P. lobata</em> may ameliorated PF via the arginine metabolism pathway in rats. Therefore, <em>P. lobata</em> may be a potential therapeutic agent to ameliorated PF.</p></div><div><h3>Conclusion</h3><p>In this work, we used metabolomics and network pharmacology to explore the mechanisms of <em>P. lobata</em> in the treatment of PF. Finally, we confirmed that <em>P. lobata</em> alleviated BLM-induced PF in rats by regulating arginine metabolism pathway based on reducing the L-OT and NOS2-related signal molecular. The search for the biomarkers finding of arginine metabolism pathway revealed a new strategy for <em>P. lobata</em> in the treatment of PF.</p></div>","PeriodicalId":15761,"journal":{"name":"Journal of ethnopharmacology","volume":"332 ","pages":"Article 118346"},"PeriodicalIF":4.8000,"publicationDate":"2024-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Integrating metabolomics and network pharmacology analysis to explore mechanism of Pueraria lobata against pulmonary fibrosis: Involvement of arginine metabolism pathway\",\"authors\":\"Hong Du , Meijuan Shao , Shangcheng Xu , Qian Yang , Jingping Xu , Hong Ke , Li Zou , Liping Huang , Yanru Cui , Fei Qu\",\"doi\":\"10.1016/j.jep.2024.118346\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Ethnopharmacological relevance</h3><p>Pueraria lobata (Willd.) Ohwi is a typical medicinal and edible plant with a long application history in China and Southeast Asia. As a widely used traditional medicine, <em>P. lobata</em> exhibits the properties of anti-inflammatory, antipyretic, antioxidant, relieving cough and asthma. Particularly, the increasing evidence indicates that the <em>P. lobata</em> has the therapeutic effect on fibrotic-related diseases in terms of metabolic regulation. However, the mechanisms of <em>P. lobata</em> on pulmonary fibrosis (PF) has not been thoroughly explored.</p></div><div><h3>Aim of the study</h3><p>This study aimed to explore the effect of arginine metabolites of <em>P. lobata</em> against PF model by integrating metabolomics and network pharmacology analysis. It might provide a new idea for the target finding of <em>P. lobata</em> anti-pulmonary fibrosis.</p></div><div><h3>Materials and methods</h3><p>In this study, the Sprague Dawley (SD) rats were randomly divided into five experimental groups: saline-treated control group, bleomycin-induced fibrosis group, prednisolone acetate group, <em>P. lobata</em> 3.2 g/kg group and <em>P. lobata</em> 6.4 g/kg group. The therapeutic effect of <em>P. lobata</em> on bleomycin-induced PF in rats was evaluated by clinical symptoms such as lung function, body weight, hematoxylin eosin staining (HE), Masson staining and hydroxyproline assay. Next, the plasma metabolomics analysis was carried out by LC-MS to explore the pathological differences between the group of control, PF and <em>P. lobata</em>-treated rats. Then, the network pharmacology study coupled with experimental validation was conducted to analysis the results of metabolic research. We constructed the “component-target-disease” network of <em>P. lobata</em> in the treatment of PF. In addition, the molecular docking method was used to verify the interaction between potential active ingredients and core targets of <em>P. lobata</em>. Finally, we tested NOS2 and L-OT in arginine-related metabolic pathway in plasma of the rats by enzyme-linked immunosorbent assay (ELISA). Real-time PCR was performed to observe the level of TNF-α mRNA and MMP9 mRNA. And we tested the expression of TNF-α and MMP9 by Western blot analysis.</p></div><div><h3>Results</h3><p>Our findings revealed that <em>P. lobata</em> improved lung function and ameliorated the pathological symptoms, such as pathological damage, collagen deposition, and body weight loss in PF rats. Otherwise, the plasma metabolomics were employed to screen the differential metabolites of amino acids, lipids, flavonoids, arachidonic acid metabolites, glycoside, etc. Finally, we found that the arginine metabolism signaling mainly involved in the regulating of <em>P. lobata</em> on the treatment of PF rats. Furtherly, the network pharmacology predicted that the arginine metabolism pathway was contained in the top 20 pathways. Next, we integrated metabolomics and network pharmacology that identified NOS2, MMP9 and TNF-α as the <em>P. lobata</em> regulated hub genes by molecular docking. Importantly, it indicated a strong affinity between the puerarin and the NOS2. <em>P. lobata</em> attenuated TNF-α, MMP-9 and NOS2 levels, suppressed TNF-α and MMP-9 protein expression, and decreased L-OT and NOS2 content in PF rats. These results indicated that the effects of <em>P. lobata</em> may ameliorated PF via the arginine metabolism pathway in rats. Therefore, <em>P. lobata</em> may be a potential therapeutic agent to ameliorated PF.</p></div><div><h3>Conclusion</h3><p>In this work, we used metabolomics and network pharmacology to explore the mechanisms of <em>P. lobata</em> in the treatment of PF. Finally, we confirmed that <em>P. lobata</em> alleviated BLM-induced PF in rats by regulating arginine metabolism pathway based on reducing the L-OT and NOS2-related signal molecular. The search for the biomarkers finding of arginine metabolism pathway revealed a new strategy for <em>P. lobata</em> in the treatment of PF.</p></div>\",\"PeriodicalId\":15761,\"journal\":{\"name\":\"Journal of ethnopharmacology\",\"volume\":\"332 \",\"pages\":\"Article 118346\"},\"PeriodicalIF\":4.8000,\"publicationDate\":\"2024-05-21\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of ethnopharmacology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0378874124006457\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"CHEMISTRY, MEDICINAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of ethnopharmacology","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0378874124006457","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, MEDICINAL","Score":null,"Total":0}
Integrating metabolomics and network pharmacology analysis to explore mechanism of Pueraria lobata against pulmonary fibrosis: Involvement of arginine metabolism pathway
Ethnopharmacological relevance
Pueraria lobata (Willd.) Ohwi is a typical medicinal and edible plant with a long application history in China and Southeast Asia. As a widely used traditional medicine, P. lobata exhibits the properties of anti-inflammatory, antipyretic, antioxidant, relieving cough and asthma. Particularly, the increasing evidence indicates that the P. lobata has the therapeutic effect on fibrotic-related diseases in terms of metabolic regulation. However, the mechanisms of P. lobata on pulmonary fibrosis (PF) has not been thoroughly explored.
Aim of the study
This study aimed to explore the effect of arginine metabolites of P. lobata against PF model by integrating metabolomics and network pharmacology analysis. It might provide a new idea for the target finding of P. lobata anti-pulmonary fibrosis.
Materials and methods
In this study, the Sprague Dawley (SD) rats were randomly divided into five experimental groups: saline-treated control group, bleomycin-induced fibrosis group, prednisolone acetate group, P. lobata 3.2 g/kg group and P. lobata 6.4 g/kg group. The therapeutic effect of P. lobata on bleomycin-induced PF in rats was evaluated by clinical symptoms such as lung function, body weight, hematoxylin eosin staining (HE), Masson staining and hydroxyproline assay. Next, the plasma metabolomics analysis was carried out by LC-MS to explore the pathological differences between the group of control, PF and P. lobata-treated rats. Then, the network pharmacology study coupled with experimental validation was conducted to analysis the results of metabolic research. We constructed the “component-target-disease” network of P. lobata in the treatment of PF. In addition, the molecular docking method was used to verify the interaction between potential active ingredients and core targets of P. lobata. Finally, we tested NOS2 and L-OT in arginine-related metabolic pathway in plasma of the rats by enzyme-linked immunosorbent assay (ELISA). Real-time PCR was performed to observe the level of TNF-α mRNA and MMP9 mRNA. And we tested the expression of TNF-α and MMP9 by Western blot analysis.
Results
Our findings revealed that P. lobata improved lung function and ameliorated the pathological symptoms, such as pathological damage, collagen deposition, and body weight loss in PF rats. Otherwise, the plasma metabolomics were employed to screen the differential metabolites of amino acids, lipids, flavonoids, arachidonic acid metabolites, glycoside, etc. Finally, we found that the arginine metabolism signaling mainly involved in the regulating of P. lobata on the treatment of PF rats. Furtherly, the network pharmacology predicted that the arginine metabolism pathway was contained in the top 20 pathways. Next, we integrated metabolomics and network pharmacology that identified NOS2, MMP9 and TNF-α as the P. lobata regulated hub genes by molecular docking. Importantly, it indicated a strong affinity between the puerarin and the NOS2. P. lobata attenuated TNF-α, MMP-9 and NOS2 levels, suppressed TNF-α and MMP-9 protein expression, and decreased L-OT and NOS2 content in PF rats. These results indicated that the effects of P. lobata may ameliorated PF via the arginine metabolism pathway in rats. Therefore, P. lobata may be a potential therapeutic agent to ameliorated PF.
Conclusion
In this work, we used metabolomics and network pharmacology to explore the mechanisms of P. lobata in the treatment of PF. Finally, we confirmed that P. lobata alleviated BLM-induced PF in rats by regulating arginine metabolism pathway based on reducing the L-OT and NOS2-related signal molecular. The search for the biomarkers finding of arginine metabolism pathway revealed a new strategy for P. lobata in the treatment of PF.
期刊介绍:
The Journal of Ethnopharmacology is dedicated to the exchange of information and understandings about people''s use of plants, fungi, animals, microorganisms and minerals and their biological and pharmacological effects based on the principles established through international conventions. Early people confronted with illness and disease, discovered a wealth of useful therapeutic agents in the plant and animal kingdoms. The empirical knowledge of these medicinal substances and their toxic potential was passed on by oral tradition and sometimes recorded in herbals and other texts on materia medica. Many valuable drugs of today (e.g., atropine, ephedrine, tubocurarine, digoxin, reserpine) came into use through the study of indigenous remedies. Chemists continue to use plant-derived drugs (e.g., morphine, taxol, physostigmine, quinidine, emetine) as prototypes in their attempts to develop more effective and less toxic medicinals.