Efficacy and potential mechanisms of jatrorrhizine on MNNG-induced chronic atrophic gastritis in rats based on serological metabolomics and molecular docking

Abstract

Jatrorrhizine (JATR), a natural isoquinoline alkaloid from Coptidis Rhizoma, exhibits various pharmacological activities, including antibacterial, anti-inflammatory, and antitumor effects. While JATR is known to treat chronic gastritis, its therapeutic potential for chronic atrophic gastritis (CAG) and its underlying mechanisms are not fully understood. This study induced CAG in rats using N-Methyl-N'-nitro-N-nitrosoguanidine (MNNG) for 12 weeks through free drinking and force-feeding. Serological metabolomics identified 23 core targets of JATR related to CAG improvement. Reverse transcription-quantitative polymerase chain reaction and western blotting confirmed the involvement of these targets. Molecular docking revealed interactions between JATR and IL-1β and Caspase-3. JATR significantly alleviated gastric inflammation and atrophy, with Kyoto Encyclopedia of Genes and Genomes analysis showing enrichment in the "Nod-like receptor-related pyroptosis pathway". JATR also enhanced GES-1 cell proliferation and reduced MNNG-induced cell damage. Additionally, JATR downregulated pyroptosis-related (Gasdermin D, NLRP3, Caspase-1) and apoptosis-related (Bcl-2, Bax, Caspase-3) markers. These findings suggest that JATR may ameliorate MNNG-induced CAG by inhibiting the activation of the Nod-like receptor-related pyroptosis pathway, supporting its potential as a therapeutic intervention for CAG.

Keywords: Caspase-3; Chronic atrophic gastritis; Jatrorrhizine; MNNG; Molecular docking; NL family pyrin domain containing 3.

Fig. 1

Metabolic profiles and differentiation of the vehicle, model, and JATR-H groups (n = 8) by multivariate analysis. The PCA score plots in ESI− mode (A) and ESI+ mode (D). PLS-DA score plots between the vehicle and model groups in ESI− mode (B) and ESI+ mode (E), between the model and JATR-H groups in ESI− mode (C) and ESI+ mode (F), (n = 8).

Fig. 2

Metabolite volcano plot of the OPLS-DA model for the vehicle and model groups in ESI− mode (A) and ESI+ mode (B), between the model and JATR-H groups in ESI− mode (C) and ESI+ mode (D). 100 permutation tests to evaluate the quality of the OPLS-DA model between the vehicle and model groups in ESI− mode (E) and ESI+ mode (F), between the model and JATR-H groups in ESI− mode (G) and ESI+ mode (H).

Fig. 3

Spearman analysis of correlation potential biomarkers and serological indices among the vehicle, the CAG group, and the JATR-H group.

Fig. 4

Visualization of gene-related information affecting endogenous DM. (A) The target genes for DM; (B) Visualized diagram of the pathway; (C) GO enrichment results of the DM-related genes. DM Differential metabolites.

Fig. 5

Effects of JATR on macroscopic pathological changes of gastric mucosa stomach tissue in CAG rats (A). Bodyweight of rats during the treatment period (B). JATR relieved histological lesions of gastric tissues in CAG rats induced by MNNG (C). Data were expressed as mean ± SD (n = 8). *P < 0.05, **P < 0.01 versus model; ^##P < 0.01 versus vehicle.

Fig. 6

JATR regulated the serological levels of gastrointestinal hormones and inflammatory cytokines in CAG rats induced by MNNG, (A) GAS-17, (B) PGI, (C) PGI/PGII, (D) IL-18, (E) IL-1β were detected using ELISA kits. The apoptosis status of gastric tissue in CAG rats was imaged histologically by TUNEL staining (F–K). Data were expressed as mean ± SD (n = 8). *P < 0.05, **P < 0.01 versus model; ^##P < 0.01 versus vehicle.

Fig. 7

Molecular docking results of JATR on IL-1β and caspase 3. (A) JATR-IL-1β. (B) JATR-caspase 3.

Fig. 8

JATR affected the pyroptosis-associated protein expression in CAG rats induced by MNNG. Cropped blots are displayed; full-length blots are provided in Supplementary Information. Data were expressed as mean ± SD (n = 8), and the experiments were repeated 3 times. *P < 0.05, **P < 0.01 versus Model; ^##P < 0.01 versus vehicle.

Fig. 9

JATR affected the expression of apoptosis-associated proteins in gastric tissues of CAG rats induced by MNNG. The expression levels of Bcl-2, Bax, and cleaved caspase-3 were measured using western blotting. Cropped blots are displayed; full-length blots are provided in Supplementary Information. Data were expressed as mean ± SD (n = 8), and the experiments were repeated 3 times. *P < 0.05, **P < 0.01 versus model; ^##P < 0.01 versus vehicle.

Fig. 10

Effect of MNNG and JATR on cell viability of GES-1 (A); Cell survival rate of GES-1 treated with different doses of MNNG for different times; (B) Cell survival rate of GES-1 treated with different doses of JATR (C). Protective effect of JATR pretreatment on proliferation of MNNG co-culturing with GES-1 cells. Histogram of NLRP3 (D), Caspase1 (E), GSDMD (F), IL-1β (H), Cyclin D1 (I), Bcl-2 (J) and Bax (K) mRNA expression in MNNG co-cultured GES-1 cells by qRT-PCR. Data were shown as mean ± SD (n = 6). *P < 0.05, **P < 0.01 versus model; ^##P < 0.01 versus vehicle.

Fig. 11

Schematic diagram of MNNG ameliorates MNNG-induced chronic gastric mucosal injury by upregulating pyroptosis and apoptosis.

Fig. 12

Experimental design of MNNG-induced CAG and drug treatment.