miRNA-Targeted Herbal Compounds Against Gastric Precancerous Evolution in Chronic Atrophic Gastritis

2.1. Search Strategy

Published studies were retrieved from electronic databases including MEDLINE, PubMed, EMBASE, and the Cochrane Library, with the search period spanning from database inception to April 25, 2026. The search strategy terms used were as follows: “gastritis, atrophic”[MeSH Terms] OR (“gastritis” AND “atrophic” OR “atrophic gastritis” OR (“chronic” AND “atrophic” AND “gastritis”) OR “chronic atrophic gastritis” OR ((“gastrics” OR “stomach”[MeSH Terms] OR “stomach” OR “gastric”) AND “Precancerous” AND (“lesion” OR “lesions” OR “lesional” OR “lesions”))) AND (“medicine, chinese traditional”[MeSH Terms] OR (“medicine” AND “chinese” AND “traditional”) OR “chinese traditional medicine” OR (“traditional” AND “chinese” AND “medicine”) OR “traditional chinese medicine”. In addition, we carried out manual retrieval of the China national knowledge infrastructure (CNKI) and the Wan Fang Database to identify publications. We also searched clinical trial registries, including ClinicalTrials.gov ( https://clinicaltrials.gov/) and Controlled-Trials.com ( www.controlled-trials.com), for ongoing studies. The search was restricted to studies published in English or Chinese. Additionally, several newly published articles were retrieved by manually reviewing the reference lists of relevant studies.

2.2. Study Criteria and Selection

Inclusion criteria for studies were applied as follows: (1) Participants: patients were diagnosed with Gastric Precancerous Evolution in Chronic Atrophic Gastritis, GC, CAG; (2) Intervention: conventional treatment + TCM, Chinese herbology, acupuncture; (3) Control group: conventional treatment; (4) Study design: Clinical trials, basic experiments, and reviews; if not published in English or Chinese, they are excluded.

5.1. Chinese Medical Herbs

While clinical trials have demonstrated the efficacy of the Huosu-Yangwei (HSYW) prescription in treating advanced GC and CAG with precancerous lesions, its molecular mechanisms remain incompletely elucidated. ³⁵ To address this gap, a systems biology approach integrating transcriptomics and network pharmacology was employed to investigate the regulatory role of HSYW’s circRNA-miRNA-mRNA (CMM) network in gastric carcinogenesis. ³⁶ The experiments conducted on Balb/c mice carrying N87 cells have verified that HSYW has an anti-tumor effect. RNA sequencing (RNA-seq) identified 200 DE mRNAs and 119 DE circRNAs, which were subsequently analyzed through predictive miRNA target interactions. Computational modeling constructed both CMM and protein-protein interaction (PPI) networks, followed by quantitative real-time PCR (qRT-PCR) validation to ensure network reliability. ³⁷ Bioinformatic interrogation of The Cancer Genome Atlas (TCGA) and Human Protein Atlas (HPA) databases revealed significant dysregulation of key differentially expressed (DE) targets, including KLF15 and PREX1. ³⁸ Core network analysis identified four circRNAs, six mRNAs, and five miRNAs as potential biomarkers for HSYW therapeutic response. Notably, the study highlights two critical regulatory axes: circRNA_07980/miR-766-3p/PREX1 and circRNA_00240/miR-642a-5p/KLF15, which may mediate HSYW’s antitumor effects in GC. These findings underscore the utility of combining experimental and computational approaches to decipher complex herbal medicine mechanisms, providing a foundation for future biomarker-driven therapeutic strategies in GC management (Figure 2 and Table1).OPEN IN VIEWER

OPEN IN VIEWER

Table 1.

Modulation of miRNA to Mitigate the Transition From CAG to GPL Using TCM

Classification	Chinese herbology	Type of study	Model	Dose/Acupoint	Effects	Refs
Chinese medical herbs	HSYW	In vivo	mice	60 mL/60 kg×0.02 kg×10.0	inhibited tumor growth	36
​	YZSJD	In vivo; In vitro	Rats, MKN-45 cells	9 g/kg	mediated by microRNA-7 on apoptosis	39
​	XLHZ	In vivo; In vitro	Rats, GES-1 cells	36 g·kg-1	altering miRNA expression profiles and regulating downstream target genes	40
​	YWXY	In vivo; In vitro	mice, human	15 g/kg, 20 g/kg	Restored the expression of miR-7 by mediating TFF2	41
​	WFC	In vitro	Human GC cell	50 mg/kg; 5 to 50 μg/mL	reducing the levels of KPNA2	42
​	Zhiwei Fuwei Pills	In vivo	Rats	0. 42 g·kg-1, 0. 84 g·kg-1, 1. 67 g·kg-1	regulating the miRNA-21/Bcl-2 signaling pathway	43
Acupuncture	NA	In vivo	Rats	Zusanli (ST36), Zhongwan (CV12), Pishu (BL20)	Decreases NF-κB p65, miR-155, miR-21 and increases miR-146a	44

HSYW, Huosu-Yangwei: Pogostemon cablin, Perilla frutescens, Atractylodes macrocephala Koidz, Citrus trifoliata, Amomum kwangsiense D, Citrus medica, Prunus mume, Zingiber officinale Roscoe, Ziziphus jujuba, Glycyrrhiza uralensis Fisch, Astragalus mongholicus Bunge, Rehmannia glutinosa, Paeoni, Trichosanthes kirilowii Maxim, Angelica sinensis, Conioselinum anthriscoides, Curcuma zedoaria, Lycium chinense, Coptis chinensis Franch, Codonopsis pilosula, Taraxacum mongolicum; YZSJD, Yangzheng Sanjie Decoction: Astragali Radix, Scutellariae Barbatae Herba, Arisaematis Rhizoma Preparatum, Citri Sarcodactylis Fructus, Cremastrae Pseudobulbus and Curcumae Longae Rhizoma; XLHZ, Xianglian Huazhuo formula: Rhizoma Coptidis, Rhizoma Atractylodis Macrocephalae, Fructus Amomi Rotundus, Herba Hedyotidis, Radix Angelicae Sinensis, Rhizoma Chuanxiong, Radix Notoginseng, Poria, Radix Paeoniae Alba, Bulbus Lilii Lancifolii, Fructus Aurantii Immaturus, Rhizoma Cyperi, Radix Aucklandiae, Scorpion, Rhizoma Corydalis Yanhusuo, Herba Agastaches Rugosa, Fructus Amomi, Herba Scutellariae Barbatae; YWXY, Yiwei Xiaoyu granules: Red Ginseng, Panax Notoginseng, Atractylodis Macrocephalae Rhizoma, Coicis Semen, Angelica APA, Aesculi Semen, Fritillariae Thunbergii Bulbus et al; WFC, Weifuchun: Radix Ginseng Rubra, Rabdosia amethystoides, Fructus Aurantii; NF-κB, nuclear factor-kappa B.

Emerging evidence highlights the pivotal regulatory role of miRNAs in the molecular pathogenesis of gastric carcinogenesis, particularly during the transition from CAG to GPL, where they function as either oncogenic drivers (oncomiRs) or tumor suppressors (tumor-suppressor-miRs). ⁴⁵ Among these, miR-7 has been identified as a key tumor-suppressive miRNA in CAG-associated precancerous transformation, exerting its inhibitory effects by directly targeting the epidermal growth factor receptor (EGFR), a well-characterized oncogenic signaling hub. Yangzheng Sanjie Decoction (YZSJD), a traditional Chinese herbal formulation, has demonstrated clinical efficacy in alleviating symptoms and potentially delaying the progression of precancerous lesions, prompting mechanistic investigations into its anti-tumor properties. ⁴⁶ To elucidate the underlying molecular mechanisms, a recent study employed a YZSJD-containing serum (YCS) model to assess its effects on the GC MKN-45 cell line. ³⁹ The findings revealed that YCS significantly inhibited cellular proliferation by inducing S-phase cell cycle arrest and promoting apoptosis, while miR-7 expression was notably downregulated in GC tissues compared to normal controls. Further Western blot and qRT-PCR analyses demonstrated an inverse correlation between miR-7 and EGFR levels, suggesting that YCS-mediated suppression of GC cell proliferation and induction of apoptosis may occur, at least in part, through miR-7-dependent EGFR downregulation. ⁴⁷ These insights provide a novel mechanistic basis for YZSJD’s therapeutic potential in GC intervention, positioning miR-7/EGFR axis modulation as a promising strategy for chemoprevention and targeted therapy in gastric carcinogenesis.

This study elucidated the molecular mechanisms underlying Xianglian Huazhuo formula (XLHZ)-mediated inhibition of CAG progression to GC. ⁴⁰ Gastric mucosal pathology was assessed in a rat model, and miRNA profiling was performed via high-throughput sequencing. DE miRNAs were analyzed using miRDB, miRanda, and miRWalk to predict target genes, followed by functional enrichment analysis (GO/KEGG). Key findings were validated by qRT-PCR, while Western blot, EdU incorporation, and flow cytometry assays evaluated XLHZ’s effects on EMT markers, proliferation, migration, and cell cycle dynamics in CAG cells. Bioinformatic analysis identified four critical target genes and five DE miRNAs. ⁴⁸ GO enrichment implicated these genes in neuronal development, transcription factor regulation, and RNA polymerase activity. KEGG pathway analysis revealed significant associations with the Phospholipase D, Wnt, and MAPK signaling cascades. Experimental validation confirmed miRNA-target gene interactions consistent with sequencing data. ⁴⁹ Functionally, XLHZ upregulated E-cadherin while suppressing transforming growth factor β1 (TGF-β1), vimentin, and β-catenin expression, thereby attenuating CAG cell migration and proliferation. Cell cycle arrest at G0/G1 and G2/M phases further substantiated XLHZ’s tumor-suppressive effects. These results suggest that XLHZ impedes CAG-to-GC progression via modulation of miR-320-3p, miR-20a-3p, miR-883-3p, miR-483-3p, and miR-34b-5p and their downstream targets. This study provides novel mechanistic insights into XLHZ’s therapeutic potential, highlighting its role in miRNA-mediated pathway regulation as a strategy to mitigate gastric carcinogenesis.

Spasmolytic polypeptide-expressing metaplasia (SPEM) is an underlying PLGC. The purpose was to elucidate the preventive impacts of Yiwei Xiaoyu granules (YWXY) mediated by miR-7 in the SPEM lesions. ⁴¹ With signed informed consent, gastric mucosal tissue was gathered from healthy individuals and CAG patients. Mice in which SPEM was induced by tamoxifen were treated with YWXY. On the tenth day of the experiment, the gastric mucosa was collected. Subsequently, confirm SPEM and lesions, and study its underlying mechanism. qRT-PCR was utilized to detect RNA transcripts. In SPEM lesions, the expression of miR-7 was reduced, and the expressions of trefoil factor 2 (TFF2) and clusterin were high. ⁵⁰ Experimental research indicated that YWXY was able to suppress cell proliferation through the regulation of Ki67. At the same time, as detected by immunofluorescence, YWXY was able to restore the expression of miR-7 by regulating TFF2, suggesting its underlying machinery of targeting miR-7 via modulating TFF2. Within 3 days of YWXY administration for the SPEM lesions, the vascular endothelial growth factor-β (VEGF-β) expression and gastric intrinsic factor recovered. This led to the speculation that YWXY hinders the progression of SPEM by modulating VEGF-β and gastric intrinsic factor as its underlying mechanism. In summary, in the human gastric mucosa, the down-regulation of miR-7 occurs early in SPEM via the modulation of TFF2. ⁵¹ In the SPEM mouse model, YWXY can suppress cell proliferation and reinstate the miR-7 expression via modulating TFF2.

Weifuchun (WFC), a TCM with established efficacy in chronic gastric disorders such as atrophic gastritis and intestinal metaplasia, exhibits potential anti-tumor properties in GC, though its underlying mechanisms remain poorly characterized. ⁵² To address this gap, this study systematically evaluated WFC’s inhibitory effects on GC oncogenicity and dissected its molecular pathways. ⁴² In vitro experiments demonstrated WFC’s selective suppression of malignant phenotypes in GC cells, with minimal impact on normal gastric epithelium. Functional assays and molecular profiling revealed that WFC attenuates tumor progression primarily through downregulation of karyopherin subunit alpha 2 (KPNA2), a nuclear transport protein implicated in oncogenesis. Mechanistically, WFC modulates KPNA2 via dual pathways: (1) miR-26a-5p upregulation, inducing post-transcriptional silencing, and (2) suppression of MAPK phosphorylation dynamics, disrupting oncogenic signaling. These findings were validated in vivo, where WFC significantly impaired tumor growth in a xenograft model. ⁵³ This study identifies WFC as a multi-targeted GC therapeutic, acting through coordinated miR-26a-5p/KPNA2 axis modulation and MAPK pathway inhibition. These insights not only advance the pharmacological understanding of WFC but also support its potential integration into GC treatment regimens, particularly for therapy-resistant cases.

The present research sought to assess how Zhiwei Fuwei Pills modulate mitochondrial apoptosis in PLGC-bearing rats, centering on the regulatory role of the miRNA-21/Bcl-2 signaling pathway. ⁴³ Male SPF SD rats aged 50 weeks were selected for this study. Seventy-five of these animals were treated with N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) plus multiple pathogenic factors to induce lesions, and a reliable PLGC rat model was achieved after 26 weeks of induction. Rats with successfully established PLGC models were randomly stratified into five groups, including the model control group, folic acid positive control group, and Zhiwei Fuwei Pills low-, medium-, and high-dose groups. The other 10 rats were assigned to the negative control group, with intragastric administration of normal saline at an equivalent volume. Rats received consecutive medication treatment over four weeks, followed by the collection of gastric mucosa samples for further laboratory assays. For the assessment of pathological and molecular modifications, multiple experimental methodologies were implemented: Hematoxylin-eosin (HE) staining was carried out to characterize the pathological features of gastric mucosal tissues; TUNEL assay was employed to determine the apoptotic index of gastric mucosal epithelial cells; RT-PCR was utilized to quantify the mRNA transcript levels of miRNA-21, PTEN, Bcl-2, Bax, and caspase-3; Western blot analysis was performed to assess the protein concentrations of PTEN, Bcl-2, Bax, and caspase-3; Immunohistochemistry (IHC) was applied to detect the positive expression patterns of PTEN, Bcl-2, and Bax in gastric mucosal tissues; and Transmission electron microscopy (TEM) was used to analyze mitochondrial morphological and structural changes. These data indicated that, in comparison to the model control group, rats administered Zhiwei Fuwei Pills presented with mitigated pathological alterations in the gastric mucosa, accompanied by a prominent upregulation in apoptotic cell numbers. ⁵⁴ Further molecular assays demonstrated that the drug intervention remarkably inhibited the mRNA expression of miRNA-21 and Bcl-2. Conversely, the transcriptional and protein levels of PTEN, Bax, and caspase-3 were greatly enhanced, accompanied by a distinct reduction in Bcl-2 protein expression in all medication groups. Notably, mitochondrial ultrastructural damages were substantially relieved in all intervention groups, showing alleviated swelling, reduced membrane breakage and attenuated cristae depletion. These changes further verified the activation of mitochondria-mediated apoptotic progression. In summary, these findings reveal that Zhiwei Fuwei Pills exert a potent pro-apoptotic effect on PLGC cells via the mitochondrial pathway, and this pharmacotherapeutic action may be orchestrated by regulating the miRNA-21/Bcl-2 signaling axis.

5.2. Acupuncture

For centuries, TCM has employed acupuncture as an effective therapeutic intervention for CAG^55,56. This study systematically investigated the therapeutic mechanisms of acupuncture at Zusanli (ST36), Zhongwan (CV12), and Pishu (BL20) in a CAG-induced rat model through comprehensive evaluation of body weight dynamics, gastric gland histopathology, and molecular expression profiles of NF-κB p65 and key microRNAs (miR-155, miR-21, miR-146a). ⁴⁴ The results demonstrated that acupuncture treatment significantly improved physiological parameters, as evidenced by marked increases in body weight compared to the model group, suggesting enhanced metabolic and nutritional status. Histopathological examinations further revealed substantial amelioration of gastric mucosal damage, indicating acupuncture’s protective effects on gastric tissue integrity. At the molecular level, the CAG model exhibited upregulated expression of pro-inflammatory mediators NF-κB p65, miR-155 and miR-21, with miR-21 showing a direct positive correlation with miR-155, while anti-inflammatory miR-146a was downregulated and inversely correlated with miR-155/miR-21. Notably, acupuncture intervention effectively reversed these pathological molecular patterns, downregulating NF-κB p65, miR-155 and miR-21 while upregulating miR-146a expression. ⁵⁷ These findings collectively suggest that acupuncture’s therapeutic benefits in CAG may be mediated through modulation of the NF-κB signaling pathway and restoration of microRNA homeostasis, highlighting the critical involvement of these molecular targets in acupuncture’s anti-inflammatory and immunomodulatory effects, thereby providing a scientific basis for its clinical application in CAG management.

6.1. Limitations and future Perspectives

Lack of standardized literature retrieval strategies and methodological quality evaluation systems renders relevant studies susceptible to publication bias; The majority of included studies are preclinical basic investigations (e.g., in vitro cell assays and animal models), whereas high-quality controlled clinical trials are rare due to small sample sizes and insufficient follow-up durations, hampering the direct translation of experimental findings to human pathological conditions; In terms of mechanism exploration, existing research is mostly limited to superficial investigation of single miRNAs and linear signaling pathways, lacking targeted binding verification assays; Additionally, it fails to fully account for confounding factors including the spatiotemporal expression specificity of miRNAs, interactive regulation of non-coding RNAs, gastric mucosal microenvironment and Helicobacter pylori infection.

Future studies should prioritize clarifying the dynamic expression characteristics of tumor-suppressive miRNAs (TS-miRNAs) specific to different gastric carcinogenesis stages, such as miR-34a, miR-145, the let-7 family and miR-200c. By integrating TCM’s holistic approach with modern molecular medicine, this ancient practice may provide a novel and effective strategy for preventing gastric cancer progression through targeted miRNA modulation. Large-sample, multicenter clinical cohorts are required to verify their dual value as early warning biomarkers for malignant transformation and therapeutic targets of TCM intervention.