Myristica fragrans water extract modulates multiple biological processes to pre-protect anhydrous ethanol-induced gastric ulcers via Akt/JNK/Nrf2 pathway activation

Ethnopharmacological relevance

Myristica fragrans (Nutmeg) is a commonly used Chinese herbal medicine and edible spice. According to Pharmacopoeia of People's Republic of China, it has the effects of warming the middle and promoting qi, astringent intestines, and antidiarrheal. In the record of Compendium of Materia Medica, it is the myristica fragrans water extract (MFWE) that is utilized for therapeutic purposes of gastrointestinal disorders frequently.

Research purpose

This study is to investigate the pharmacodynamic material foundation and molecular mechanism of myristica fragrans on gastric ulcers using UHPLC-Q-Orbitrap-MS/MS with network pharmacology and experimental verification. This may provide theoretical guidance for the clinical use of myristica fragrans, and support a theoretical foundation for its future advancement into natural functional products that can relieve acute gastric ulcers.

Materials and methods

Using UHPLC/MS technology and network pharmacology, we identified possible active chemicals molecules, screened out core targets and core pathways, and simulated drug target binding through molecular docking situations. Acute gastric ulcer was caused by intragastric administration of absolute ethanol (0.075 ml/10g). Myristica fragrans water extract (182 mg/kg and 364 mg/kg) was administered orally 14 days in advance. The same method was used to distribute 0.5% carboxymethyl cellulose solution into the Model and Control group. The mice were murdered on the 15th day. Following the sacrifice, the gastric tissue was removed for histological analysis. The tissue needs to detect levels of IL-1β, TNF-α, IL-10, and IL-6 as well as the activity of SOD, GSH-Px, MDA, and MPO. In addition, H&E staining and the TUNEL method were used to observe the effect on the gastric mucosa of mice. Western blot was used to detect apoptosis, ferroptosis, and antioxidation-related proteins.

Results

A total of 10 chemical constituents were identified from MFWE using UHPLC-Q-Orbitrap MS/MS and TCMSP database. Through the network pharmacological analysis of these identified components, it was discovered that the protective effect is mainly carried out by six compounds, they are: Myristicin, Myrisligna, Ferulaldehyde, Dehydrodiisoeugenol, 7-Methoxy-4-methylcoumarin, 1,5-Bis(2,5-dimethoxyphenyl) pentane-1,5-dione. Furthermore, MFWE was found to significantly reduce TNF-α, IL-1β, and IL-6, increase IL-10, and alleviate the inflammation caused by alcoholic gastric ulcers. It can lower MDA and MPO, raise SOD and GSH-Px to relieve oxidative stress. Results from network pharmacology indicated that the Akt, JNK, and apoptosis signaling pathways were essential for the therapeutic effects of MFWE on gastric ulcers. Further literature research revealed that Nrf2 and ferroptosis signaling pathways may be related to the role of MFWE. Molecular biology studies confirmed that MFWE decreased the expression levels of p-Akt/Akt, p-JNK/JNK, Bax, and Keap-1, and increased the expression levels of Bcl-2, Nrf2, HO-1, SLC7A11, GPX4, FTH1.

Conclusion

This study demonstrates that MFWE can alleviate inflammatory responses and diminish both cellular apoptosis and ferroptosis. they confer a protective effect on gastric ulcers via the activation of the Akt/JNK-Keap1-Nrf2-HO-1 signaling pathway and offer a promising therapeutic strategy.