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| Imperatorin promotes the browning of white fat through the AMPK/SIRT1/PGC-1α pathway |
| Hits 131 Download times 38 Received:November 20, 2025 |
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| DOI
10.11656/j.issn.1673-9043.2026.02.07 |
| Key Words
network pharmacology;imperatorin;thermogenesis;white adipose tissue browning;survival time |
| Author Name | Affiliation | E-mail | | BAI Xia | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Military Medical Sciences Academy, Tianjin 300050, China | | | ZHANG Guanyu | Military Medical Sciences Academy, Tianjin 300050, China | | | WU Shuai | Military Medical Sciences Academy, Tianjin 300050, China | | | ZHANG Yongqiang | Military Medical Sciences Academy, Tianjin 300050, China | | | ZHANG Li | Military Medical Sciences Academy, Tianjin 300050, China | | | YANG Danfeng | Military Medical Sciences Academy, Tianjin 300050, China | | | LI Xi | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Military Medical Sciences Academy, Tianjin 300050, China | woshiliulangdeyu@163.com |
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| Abstract
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| [Objective] To screen traditional Chinese medicine（TCM）monomers that promote white adipose tissue browning using network pharmacology，evaluate their activity，and investigate their mechanism of action. [Methods] 1）Screening of TCM monomers：GEO2R was used to analyze differential genes in subcutaneous white adipose tissue of C57BL / 6 mice after one week and five weeks of cold exposure，and intersecting differentially expressed genes（DEGs）were screened. A protein-protein interaction network was constructed to identify core （Hub）genes. The ITCM platform predicted TCM monomers capable of regulating hub genes. 3T3-L1 preadipocytes were induced to differentiate with 10 μmol/L of different drugs，and real-time quantitative PCR（RT-PCR）was used to screen for the TCM monomer with the strongest up-regulatory effect on thermogenic gene expression. 2）Biological activity evaluation and mechanism study of the TCM monomer：After intervention with the optimal monomer，cell viability was detected by CCK-8 assay；lipid accumulation was observed by Oil Red O staining；uncoupling protein 1 (UCP1) expression was quantified by immunofluorescence；mitochondrial number was detected using MitoTracker Red probe；changes in browning gene and protein levels were measured by RT-PCR and Western blot；the AMP- activated protein kinase（AMPK） inhibitor dorsomorphin was added to verify whether the monomer exerts thermogenic effects by activating the AMPK/SIRT1/PGC-1α pathway. Eight-week-old male C57BL/6 mice were divided into control，low-dose，high-dose，and 2-week cold exposure groups. After 28 days of administration，orbital blood was collected to measure serum total cholesterol（TC），triacylglycerol（TG），low-density lipoprotein cholesterol (LDL-C)，and high-density lipoprotein cholesterol（HDL-C）levels. Inguinal white adipose tissue(iWAT) was isolated；histomorphology was observed by hematoxylin-eosin（HE）staining and UCP1 immunohistochemistry；gene and protein expression of related pathways was determined by RT-PCR and Western blot；a parallel group was exposed to-15 ℃ to determine survival time. [Results] Among the five TCM monomers screened by network pharmacology，imperatorin（IMP）showed the strongest up-regulatory effect on thermogenic gene expression. The optimal concentration of IMP was 10 μmol/L as determined by CCK-8 assay. After IMP intervention，lipid droplet content in adipocytes decreased，while UCP1 expression and mitochondrial number were significantly up-regulated （P<0.01）. Mitochondrial biogenesis genes nuclear respiratory factor 1（Nrf1），mitochondrial transcription factor A （Tfam），and beige-specific genes Cited1，Cd137，Tmem26，and Dio2 were significantly up-regulated（P<0.05）. At concentrations of 1，3，and 10 μmol/L，IMP dose-dependently promoted the expression of thermogenic genes UCP1， PRDM16，PGC-1α（P<0.05），thermogenic and fatty acid oxidation protein PPARα，and AMPK pathway proteins p- AMPK and SIRT1，while inhibiting the expression of lipid synthesis genes PPARγ and C/EBP. The AMPK inhibitor blocked IMP’s effects on adipose browning. After IMP intervention，mice had lower TG，TC，and LDL-C，and higher HDL-C than the control group. Lipid droplet morphology in iWAT was relatively smaller；UCP1-positive cell number significantly increased in the high-dose group（P<0.01）；high-dose IMP significantly up-regulated UCP1，PGC-1α gene and protein expression，and AMPK pathway protein expression，while down-regulating PPARγ protein expression（P<0.05）. Low-dose IMP significantly extended mouse survival time at -15 ℃（P<0.01）. [Conclusion] 1）Network pharmacology can predict TCM monomers that promote adipocyte browning. 2）IMP can promote white adipose tissue browning and effectively extend mouse survival time under extreme cold. 3）IMP promotes white adipose tissue browning by regulating the AMPK/SIRT1/PGC-1α signaling pathway. |
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