Integrated Approaches Revealed the Therapeutic Mechanisms of Zuojin Pill Against Gastric Mucosa Injury in a Rat Model with Chronic Atrophic Gastritis

Abstract

Background

The Zuojin Pill (ZJP) is widely used for treating chronic atrophic gastritis (CAG) in clinical practice, effectively ameliorating symptoms such as vomiting, pain, and abdominal distension in patients. However, the underlying mechanisms of ZJP in treating CAG has not been fully elucidated.

Purpose

This study aimed to clarify the characteristic function of ZJP in the treatment of CAG and its potential mechanism.

Methods

The CAG model was established by alternant administrations of ammonia solution and sodium deoxycholate, as well as an irregular diet. Therapeutic effects of ZJP on body weight, serum biochemical indexes and general condition were analyzed. HE staining and AB-PAS staining were analyzed to characterize the mucosal injury and the thickness of gastric mucosa. Furthermore, network pharmacology and molecular docking were used to predict the regulatory mechanism and main active components of ZJP in CAG treatment. RT-PCR, immunohistochemistry, immunofluorescence and Western blotting were used to measure the expression levels of apoptosis-related proteins, gastric mucosal barrier-associated proteins and PI3K/Akt signaling pathway proteins.

Results

The results demonstrated that ZJP significantly improved the general state of CAG rats, alleviated weight loss and gastric histological damage and reduced the serum biochemical indicators. Network pharmacology and molecular docking found that ZJP in treating CAG by inhibiting inflammation, suppressing apoptosis, and protecting the gastric mucosal barrier via the PI3K/Akt signaling pathway. Further experiments confirmed that ZJP obviously modulated the expression of key proteins involved in gastric mucosal cell apoptosis, such as Bax, Bad, Apaf-1, cleaved-caspase-3, cleaved-caspase-9, Cytochrome C, Bcl-2, and Bcl-xl. Moreover, ZJP significantly reversed the protein expression of Occludin, ZO-1, Claudin-4 and E-cadherin.

Conclusion

Our study revealed that ZJP treats CAG by inhibiting the PI3K/Akt signaling pathway. This research provided a scientific basis for the rational use of ZJP in clinical practice.

Keywords:

• Zuojin Pill
• chronic atrophic gastritis
• network pharmacology
• molecular docking
• PI3K/Akt signaling pathway

Abbreviations

ZJP, Zuojin Pill; IL-6, Interleukin-6; CAG, Chronic atrophic gastritis; IL-1β, Interleukin-1β; TCM, Traditional Chinese medicine; TNF-α, Tumor necrosis factor-α; PG I, Pepsinogen I; OB, Oral bioavailability; PG II, Pepsinogen II; DL, Drug-likeness; GAS-17, Gastrin17; RIPA, Radioimmunoprecipitation assay; KEGG, Kyoto Encyclopedia of Genes and Genomes; PMSF, Phenylmethylsulfonyl fluoride; GO, Gene Ontology.

Ethics Statement

All animal experiments were conducted in accordance with the Laboratory Care and Use Guidelines. The research was approved by Ethics Committee of the Chinese PLA General Hospital (Approval ID: IACUC-2021-0022).

Acknowledgments

This research was financially supported by the Major Program of the National Natural Science Foundation of China (No. 82192915), the National Key Research and Development Program (No. 2018YFC1704500) and the China Postdoctoral Science Foundation (No. 2023M730115).

Author Contributions

All authors made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis and interpretation, or in all these areas; took part in drafting, revising or critically reviewing the article; gave final approval of the version to be published; have agreed on the journal to which the article has been submitted; and agree to be accountable for all aspects of the work.

Disclosure

The authors declare that they have no competing interests in this work.