Targeting AGE-RAGE Signaling Pathway with Hujin Decoction Ameliorates MAFLD in HepG2 Cells

Zixuan Zhang,1,* Jiaxi Shi,2,* Fuxuan Liu,1 Jing Zhou,1 Qi Shen,3 Xuguang Shi1 1School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, 510006, People’s Republic of China; 2The First School of Clinical Medicine, Guangzhou University of Chinese Medicine, Guangzhou, 510405, People’s Republic of China; 3Science and Technology Innovation Center, Guangzhou University of Chinese Medicine, Guangzhou, 510405, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xuguang Shi, School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, People’s Republic of China, Tel +8613602740975, Email [email protected]: To explore the mechanism and substance basis of HJD for the treatment of MAFLD based on system pharmacology.Patients and Methods: The ingredients of HJD in vitro and in vivo were detected by UPLC-MS/MS, then network pharmacology and molecular docking technology were used to predict the mechanism and substance basis, then the establishment of in vitro MAFLD model was confirmed by oil red O staining and ELISA technology, and finally the mechanism was verified by PCR, WB and flow cell technology.Results: System pharmacology determined that succinic acid, Ginsenoside Rh4, Caffeic acid, 7-Methoxycoumarin, 5-Acetylsalicylic acid and other ingredients were the basis of pharmacodynamic substances, while RAGE[Advanced glycosylation end product-specific receptor (RAGE)], BCL2[Apoptosis regulator Bcl-2 (BCL2)], and CASP3[Caspase-3 (CASP3)] were predicted as the core targets, and AGE-RAGE was the key pathway. In vitro experiments confirmed that HJD can reduce hepatocyte apoptosis by downregulating the AGE-RAGE signaling pathway to alleviate MAFLD.Conclusion: HJD may act on RAGE, BCL2, CASP3, and other key targets to regulate the AGE-RAGE signaling pathway through succinic acid, Ginsenoside Rh4 and Caffeic acid. This study provides a theoretical basis for the clinical application and quality control of HJD.Keywords: Hujin decoction, MAFLD, UPLC-MS/MS, molecular docking, network pharmacology, AGE-RAGE pathway