Elucidating the Mechanism of Moluodan in Treating Chronic Non-Atrophic Gastritis: An Integrated Approach Combining Clinical Proteomics, Network Pharmacology, and Experimental Validation

Yuetian Xu¹Feiyue Zhao^2,3He Jiang²Qichao Liu²Menglei Wang^3,4Pengchao Bai²Huiyun Liu^2,3*Ruikang Jia^2,3*

• ¹Department of Gastroenterology, Affiliated Hospital of Hebei University of Engineering, Handan, China
• ²Handan pharmaceutical co. LTD, Handan, China
• ³Key Laboratory of Chinese Medicine for Gastric Medicine, Handan, China
• ⁴Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, China

Background: In China, a considerable proportion of individuals suffering from stomach ailments endure a prolonged, recurring, and difficult-to-manage disease course. Chronic non-atrophic gastritis (CNAG), a prevalent digestive tract disorder, requires robust clinical pharmacological intervention for effective treatment. Moluodan (MLD), a traditional Chinese medicine consisting of 18 herbs, is commonly utilized for chronic gastritis. However, the mechanism of its action in CNAG treatment remains unclear. Materials and Methods: Quantitative proteomic analysis was performed on pathological samples from patients diagnosed with clinical CNAG, through which differentially expressed proteins were identified and bioinformatic analyses were performed. Network pharmacology and molecular docking techniques werewas utilized to thoroughly investigate the potential targets of MLD in the treatment of CNAG. The intoblood ingredients of MLD were studied based on animal experiments, and the targets of intoblood ingredients were identified by network pharmacology. At the cell level, the anti-inflammatory effects of MLD formula and seven active ingredients were investigated. Ultimately, the gastric mucosal protective effects of MLD and its seven active ingredients were tested in an ethanol induced gastric injury model. Results: Proteomic results from clinical samples showed that 270 DEPs were obtained in CNAG disease. The network pharmacology results of MLD showed that MAPK8 (JNK) is a potential targetkaempferol could interact with MAPK8. A total of 116 intoblood ingredients of MLD were identified, and the network pharmacology of these intoblood ingredients also showed that MAPK8 (JNK) is a potential targetraffinose could interact with MAPK8. The results of cell experiments showed that MLD exhibits potent anti-inflammatory effect. The active ingredients Ginsenoside Rg1, Pseudo ginsenoside RT1, Pinicolic acid, Paeonoside, Allocryptopine, Atractyloside A, and Ferulic acid inhibited the expression of p-JNK, as well as IL-6 and IL-1β mRNA levels. In the ethanol induced gastric injury model, MLD and its seven active ingredients have potential gastric mucosal protective effects. Conclusion: The material basis of MLD treatment of CNAG may be Ginsenoside Rg1, Pseudo ginsenoside RT1, Pinicolic acid, Paeonoside, Allocryptopine, Atractyloside A, and Ferulic acid, which exert gastric mucosal protective effects by inhibiting overactivation within the JNK/p-JNK signaling pathway to inhibit cell inflammation.